(Vol.61 No.12 1997)
Progressive Effects of Phloridzin on Melanogenesis in B16
Mouse Melanoma Cells
Toshihiko Shoji, Masuko Kobori, Hiroshi Shinmoto, Masayuki Tanabe, and
Tojiro Tsushida 1963
In Vivo Antioxidant Activity of Okara Koji, a Fermented
Okara, by Aspergillus oryzae
Masako Matsuo 1968
Effects of Metal Ions and pH on the Stability of Linoleic
Acid Hydroperoxide in the Water Phase
Tamako Nishiike, Satoko Kondo, Tamae Yamamoto, Aya Shigeeda, Yoshimi Yamamoto,
Hitoshi Takamura, and Teruyoshi Matoba 1973
Flavor Characteristics of Glutathione in Raw and Cooked
Foodstuffs
Yoichi Ueda, Muneaki Yonemitsu, Takako Tsubuku, Makoto Sakaguchi, and Ryuichi
Miyajima 1977
Dose-dependent Incorporation of Tea Catechins, (-)-Epigallocatechin-3-gallate
and (-)-Epigallocatechin, into Human Plasma
Kiyotaka Nakagawa, Shiho Okuda, and Teruo Miyazawa 1981
Expression of Genes Encoding Membrane-bound Hydrogenase
in Pseudomonas hydrogenovora under Autotrophic Condition Is Dependent on
Two Different Promoters
Takashi Ohtsuki, Makoto Shimosaka, and Mitsuo Okazaki 1986
Synthesis of Didocosahexaenoylphosphatidylserine
Ryuichi Nakashima, Kouichi Kado, Takeshi Kume, and Kazuyuki Maekawa 1991
cDNA Cloning and Gene Expression of Phenylalanine Ammonia-Lyase
in Lithospermum erythrorhizon
Kazufumi Yazaki, Mieko Kataoka, Gisho Honda, Klaus Severin, and Lutz Heide
1995
Amino Acid Sequence and Stereoselective Hydrolytic Reaction
of an Endo-1,4--glucanase from a Bacillus Strain
Jun Hitomi, Yuji Hatada, Shunro Kawaminami, Shuji Kawai, and Susumu Ito
2004
Transxylosylation of -Xylosidase from Aspergillus awamori
K4
Masahiro Kurakake, Shinji Osada, and Toshiaki Komaki 2010
Effects of a Hydrogenated Isomaltooligosaccharide Mixture
on Glucan Synthesis and on Caries Development in Rats
Jun Tsunehiro, Takashi Matsukubo, Masao Shiota, and Yoshinori Takaesu 2015
Chiral Synthesis of the BC-Ring System of Ciguatoxin from
D-Glucose
Eiichi Ami, Hisakazu Kishimoto, Hiroshi Ohrui, and Hiroshi Meguro 2019
New Physiological Effects of 5-Aminolevulinic Acid in
Plants: The Increase of Photosynthesis, Chlorophyll Content, and Plant
Growth
Yasushi Hotta, Tohru Tanaka, Hideo Takaoka, Yasutomo Takeuchi, and Makoto
Konnai 2025
Continuous Production of L-Alanine with NADH Regeneration
by a Nanofiltration Membrane Reactor
Shu-Su Lin, Osato Miyawaki, and Kozo Nakamura 2029
Superoxide Dismutase Inhibition of Oxidation of Ubiquinol
and Concomitant Formation of Hydrogen Peroxide
Tsutomu Nakayama, Miku Hashimoto, and Kei Hashimoto 2034
Inactivation of Cu,Zn-Superoxide Dismutase by Intermediates
of Maillard Reaction and Glycolytic Pathway and Some Sugars
Hiroyuki Ukeda, Yuko Hasegawa, Toshinao Ishi, and Masayoshi Sawamura 2039
Synthesis and Biological Activities of 8'-Methylene- and
8'-Methylidyneabscisic Acids
Yasushi Todoroki, Sei-chi Nakano, Sho Arai, Nobuhiro Hirai, and Hajime
Ohigashi 2043
Purification and Characterization of N-Acetylneuraminate
Synthase from Escherichia coli K1-M12
Eriko Komaki, Yasuhiro Ohta, and Yoji Tsukada 2046
Modulation of GABA Receptors Expressed in Xenopus Oocytes
by 13-L-Hydoxylinoleic Acid and Food Additives
Hitoshi Aoshima and Yukihiro Tenpaku 2051
Characterization of Styrene Oxide Isomerase, a Key Enzyme
of Styrene and Styrene Oxide Metabolism in Corynebacterium sp.
Nobuya Itoh, Kunimasa Hayashi, Keisaku Okada, Takeshi Ito, and Naoyuki
Mizuguchi 2058
Glycogen-Surfactant Complexes: Phase Behavior in a Water/Phytoglycogen/Sodium
Dodecyl Sulfate (SDS) System
Akihiro Nakano, Ryozo Irie, and Koichi Tateishi 2063
Autoxidation Reaction Mechanism for L-Ascorbic Acid in
Methanol without Metal Ion Catalysis
Noriko Miyake, Yuzuru Otsuka, and Tadao Kurata 2069
Molecular Cloning of Levan Fructotransferase Gene from
Arthrobacter nicotinovorans GS-9 and Its Expression in Escherichia coli
Katsuichi Saito, Atsushi Yokota, and Fusao Tomita 2076
Effect of Polyphenol Oxidase on Deodorization
Osamu Negishi and Tetsuo Ozawa 2080
Oxidative Stability of Docosahexaenoic Acid-containing
Oils in the Form of Phospholipids, Triacylglycerols, and Ethyl Esters
Jin-Hyang Song, Yoshikazu Inoue, and Teruo Miyazawa 2085
Effects of Dietary Lipid Peroxidation Products on Hormonal
Responses in Primary Cultured Hepatocytes of Rats
Hitoshi Ashida, Kenichi Ohue, Kazuki Kanazawa, and Gen-ichi Danno 2089
Identification of the Essential Amino Acid Residues in
LukS for the Hemolytic Activity of Staphylococcal Leukocidin towards Rabbit
Erythrocytes
Hirofumi Nariya, Akiko Shimatani, Toshio Tomita, and Yoshiyuki Kamio
2095
Note
Purification and Some Properties of a Protease from the Sarcocarp of Musk
Melon Fruit
Makoto Kaneda, Hiroo Yonezawa, and Tetsuya Uchikoba 2100
Note
Synthesis and Fungicidal Activity of 6-Alkyl Six-membered Cyclic Thiophosphates
Shinkichi Tawata, Shigehiko Taira, Hirofumi Kikizu, Naotada Kobamoto, Masanobu
Ishihara, and Seizen Toyama 2103
Note
Superoxide-scavenging and Tyrosinase-inhibitory Activities of the Extracts
of Some Chinese Medicines
Zhuang Miao, Hiroshi Kayahara, and Koji Tadasa 2106
Note
Formation of Thioxopyrrolidines and Dithiocarbamates from 4-Methylthio-3-butenyl
Isothiocyanates, the Pungent Principle of Radish, in Aqueous Media
Hiroki Matsuoka, Yoshinori Toda, Kenji Yanagi, Asaka Takahashi, Koichi
Yoneyama, and Yasushi Uda 2109
Note
Diurnal Fluctuation in the Enzyme Activity and the Messenger RNA Level
of Pineal Serotonin N-Acetyltransferase in Normal and Hereditary Microphthalmic
Rats
Suhyeun Shim, Zhengwei Fu, Hisanori Kato, and Hideyuki Tanaka 2113
Note
The Effects of Niacin on DNA Repair after N-Methyl-N'-nitro-N-nitrosoguanidine
Treatment in Normal Human Lymphocytes
Shin Ogata, Katsuzumi Okumura, and Hiroshi Taguchi 2116
Note
Regulatory Region of Expression of Thiobacillus ferrooxidans leuB Gene
in Escherichia coli
Hiroshi Kawaguchi, Kenji Inagaki, and Hidehiko Tanaka 2119
Note
Starvation-increased Insulin-dependent Tyrosine Phosphorylation of the
195-kDa Protein in Intact Rat Liver
Yoshiaki Ito, Shin-Ichiro Takahashi, Asako Takenaka, Tomomi Hidaka, and
Tadashi Noguchi 2122
Note
Periplasmic Secretion of Functional Ovotransferrin N-Lobe in Escherichia
coli
Tsuguo Miyauchi, Nobuyuki Takahashi, and Masaaki Hirose 2125
Note
Eicosanyl p-Coumarates from a Kenyan Plant, Psiadia punctulata : Plant
Growth Inhibitors
Joseph M. Keriko, Shuhei Nakajima, Naomichi Baba, and Junkichi Iwasa 2127
Note
Production of New Restriction Endonuclease VpaK11BI Recognizing Sequence
5'-GGWCC-3' in a Haemolysin-less Mutant of Vibrio parahaemolyticus
Harumi Ueno, Akihiko Kita, Michiko Miyahara, Naoko Ishiwata, Katsutoshi
Mise, Ikunoshin Kato, and Yoshizumi Ishino 2129
Note
Mutational Analysis of the Potato Virus Y 5' Untranslated Region for Alteration
in Translational Enhancement in Tobacco Protoplasts
Li Jun Yang, Makoto Hidaka, Junichiro Sonoda, Haruhiko Masaki, and Takeshi
Uozumi 2131
Note
Conversion from Tryptophan Precursor into Violacein Pigments by a Cell-free
System from Chromobacterium violaceum
Tsutomu Hoshino and Masahiro Yamamoto 2134
Note
Effects of Carnosine and Anserine on the Destruction of Vitamin B(/)12
with Vitamin C in the Presence of Copper
Shigeo Takenaka, Sumi Sugiyama, Fumio Watanabe, Katsuo Abe, Yoshiyuki Tamura,
and Yoshihisa Nakano 2137
Note
Purification and Characterization of Two Chitinase Isoforms from the Bulbs
of Gladiolus (Gladiolus gandavensis)
Takeshi Yamagami, Yoichiro Mine, Yoichi Aso, and Masatsune Ishiguro 2140
Note
Molecular Cloning and Characterization of a cDNA Encoding Early Light-inducible
Protein from Soybean (Glycine max L.)
Hiroshi Yamagata and Chris Bowler 2143
Short Communication
Flutolanil Resistance as a Genetic Marker of Coprinus cinereus Strains
Shigeru Oita, Tsutomu Fushimi, Tetsuya Ookura, Yasuhiro Ito, and Sonoe
O. Yanagi 2145
Short Communication
Substantially Complete Removal of Three Major Allergenic Soybean Proteins
(Gly m Bd 30K, Gly m Bd 28K, and the -Subunit of Conglycinin) from Soy
Protein by Using a Mutant Soybean, Tohoku 124
Masahiko Samoto, Yoichi Fukuda, Koji Takahashi, Kohsei Tabuchi, Miki Hiemori,
Hideaki Tsuji, Tadashi Ogawa, and Yukio Kawamura 2148
Short Communication
A Novel Modification of the Lysine Residue at Position 12 of Histone H4
in Starfish Sperm
Kazuto Nunomura, Takahiko Shimizu, Keiko Hozumi, Toshifumi Takao, Yasutsugu
Shimonishi, and Susumu Ikegami 2151
Short communication
Simplicissin, a New Pollen Growth Inhibitor Produced by the Fungus, Penicillium
cf. simplicissimum (Oudemans) Thom No. 410
Miyako Kusano, Hiroyuki Koshino, Jun Uzawa, Shozo Fujioka, Tsuyoshi Kawano,
and Yasuo Kimura 2153
-1-
Progressive Effects of Phloridzin on Melanogenesis in B16
Mouse Melanoma Cells
Toshihiko Shoji, Masuko Kobori,* Hiroshi Shinmoto,* Masayuki Tanabe, and Tojiro Tsushida*
Institute for Production Research and Development, The Nikka Whisky
Distilling Co., Ltd., 967 Matsuyama, Masuo, Kashiwa, Chiba 277, Japan
* National Food Research Institute, Ministry of Agriculture, Forestry,
and Fisheries, 1-2-1 Kannondai, Tsukuba, Ibaraki 305, Japan
Received February 24, 1997
@When we studied the effects of polyphenols from apple fruits on melanogenesis
in B16 mouse melanoma cell lines, phloridzin had dose-dependent progressive
effects on melanogenesis between 10 and 500 g/ml without inhibiting cell
growth. At a concentration of 500 g/ml, phloridzin increased the melanin
content in the cells to 181% of that in control cells. In contrast, phloretin,
the aglycon of phloridzin, did not activate melanogenesis in the cells
and was cytotoxic at a concentration of 5 g/ml. Phloridzin increased
the activity of tyrosinase to 223% of that in control cells. Furthermore,
phloridzin inhibited the activity of protein kinase C (PKC), which is recognized
to regulate tyrosinase activity. The inhibition of PKC activity continued
for 120 min from the addition of phloridzin. Therefore, we estimated that
the activation of melanogenesis by phloridzin resulted from the increase
of tyrosinase activity caused by the inhibition of PKC activity.
Key words: phloridzin; melanogenesis; B16 mouse melanoma cell; tyrosinase;
protein kinase C
-2-
In Vivo Antioxidant Activity of Okara Koji, a Fermented
Okara, by Aspergillus oryzae
Masako Matsuo
Gifu Woman's University, 80 Taromaru, Gifu 501-25, Japan
Received March 3, 1997
@The antioxidants in Okara Koji (OK), an okara (OC) fermented by Aspergillus
oryzae, -tocopherol, -tocopherol, genistin, daizein, genistein, and
3-hydroxyanthranilic acid were identified by HPLC. OK's extract with 80%
methanol strongly inhibited linoleate peroxidation, much more than other
OK's extracts with hexane or hot water. The methanol extract accelerated
12-hydroxyeicosatetraenoic acid formation in membrane lipids at 10-3 concentration,
but inhibited the formation at higher concentrations than 10-3 ex vivo.
To confirm the total effect of all components of OK on lipid peroxidation
in vivo, rats fed food deficient in vitamin E were put on diets containing
OK or OC with oxidized oil. In rats fed the OK diet, no effect of oxidized
oil feeding on the body weight gain, of the TBA value in plasma, or of
glutathione peroxidase activities of plasma and liver was observed. But
in rats fed the OC diet, the effect of oxidized oil feeding was apparently
observed on all of those values. These results suggested that OK would
scavenge lipid peroxides in vivo.
Key words: okara; okara koji; antioxidant activity
-3-
Effects of Metal Ions and pH on the Stability of Linoleic
Acid Hydroperoxide in the Water Phase
Tamako Nishiike, Satoko Kondo,* Tamae Yamamoto,* Aya Shigeeda,* Yoshimi Yamamoto,* Hitoshi Takamura,* and Teruyoshi Matoba*
Division of Human Life and Environmental Sciences, Graduate School of
Human Culture, Nara Women's University, Kitauoya-Nishimachi, Nara 630,
Japan
* Department of Food Science and Nutrition, Nara Women's University, Kitauoya-Nishimachi,
Nara 630, Japan
Received March 27, 1997
@We examined linoleic acid hydroperoxide (hydroperoxyoctadecadienoic acid;
HPOD) decomposition kinetically with or without various metal ions and
at various pHs as effective factors on the stability of hydroperoxides.
@HPOD decomposition in the reaction system of this experiment was a first-order
reaction. Manganese, copper, and especially iron accelerated the decomposition
of HPOD, while lithium, sodium, potassium, magnesium, calcium, and aluminium
stabilized HPOD. Besides, HPOD was comparatively stable at pH 3, 7, and
8, and unstable at pH 2, 4-6, and 9. According to activation energy, however,
it was estimated that only in the reaction system with iron or at pH 2
and 9 the HPOD decomposition mechanism was different from that in water.
Key words: lipid oxidation; linoleic acid; hydroperoxide; metal ions; pH
-4-
Flavor Characteristics of Glutathione in Raw and Cooked
Foodstuffs
Yoichi Ueda, Muneaki Yonemitsu, Takako Tsubuku, Makoto Sakaguchi, and Ryuichi Miyajima
Food Research & Development Laboratories, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki 210, Japan
Received April 7, 1997
@The flavor of glutathione (-L-glutamyl-L-cysteinylglycine, GSH) was
examined by several sensory evaluations. The measurement of a point of
subjective equality (PSE) showed that the peptide increases the flavor
characteristics but did not affect the intensity of basic tastes, such
as sweetness, saltiness, sourness, and umami. However, the threshold value
of GSH decreased significantly in an umami solution containing 0.05% each
of monosodium glutamate (MSG) and disodium inosinate (IMP). This suggests
that GSH interacts with the umami substance and has a certain effect on
the flavor.
@GSH had a characteristic kokumi flavor, such as continuity, mouthfulness,
and thickness in the umami solution as well as in a model beef extract
constructed from analyzed components at a concentration of 0.02% w/v. Some
foodstuffs, including meat, were found to contain GSH above its threshold
value, which implicates the contribution of GSH to the flavor.
@The thermal degradation study suggested that a part of GSH have changed
into its disulfide, pyroglutamic acid (PCA), and cyclocysteinylglycine
in cooked foodstuffs.
Key words: glutathione; flavor characteristics; kokumi flavor; food; thermal
degradation
-5-
Dose-dependent Incorporation of Tea Catechins, (-)-Epigallocatechin-3-gallate
and (-)-Epigallocatechin, into Human Plasma
Kiyotaka Nakagawa, Shiho Okuda, and Teruo Miyazawa
Food Chemistry Laboratory, Faculty of Agriculture, Tohoku University, Sendai 981, Japan
Received April 8, 1997
@Tea catechins, (-)-epigallocatechin-3-gallate (EGCg) and (-)-epigallocatechin
(EGC), have been reported to suppress oxidation of plasma low density lipoprotein
(LDL) in vitro. If dietary catechins can be efficiently incorporated into
human blood plasma, anti-atherosclerotic effects in preventing oxidative
modification of LDL would be expected. In this study, a newly developed
chemiluminescence detection-high pressure liquid chromatography (CL-HPLC)
method for measuring plasma catechins was used and the incorporation of
EGCg and EGC into human plasma was investigated. Healthy subjects orally
ingested 3, 5, or 7 capsules of green tea extract (corresponding to 225,
375, and 525 mg EGCg and 7.5, 12.5, and 17.5 mg EGC, respectively). The
plasma EGCg and EGC concentrations before the administration were all below
the detection limit (<2 pmol/ml), but 90 min after, significantly and
dose-dependently increased to 657, 4300, and 4410 pmol EGCg/ml, and 35,
144, and 255 pmol EGC/ml, in the subjects who received 3, 5, and 7 capsules,
respectively. Both EGCg and EGC levels detected in plasma corresponded
to 0.2-2.0% of the ingested amount. Catechin intake had no effect on the
basal level of endogenous antioxidants (-tocopherol, -carotene, and
lycopene) or of lipids in plasma. These results suggested that drinking
green tea daily would contribute to maintain plasma catechin levels sufficient
to exert antioxidant activity against oxidative modification of lipoproteins
in blood circulation systems.
Key words: epigallocatechin gallate; epigallocatechin; absorption; plasma;
human
-6-
Expression of Genes Encoding Membrane-bound Hydrogenase
in Pseudomonas hydrogenovora under Autotrophic Condition Is Dependent on
Two Different Promoters
Takashi Ohtsuki,* Makoto Shimosaka,** and Mitsuo Okazaki*,**,
* Department of Applied Biology, Faculty of Textile Science and Technology, and ** Gene Research Center, Shinshu University, 3-15-1 Ueda, Nagano 386, Japan
Received April 11, 1997
@Expression of a membrane-bound hydrogenase of the hydrogen-utilizing
bacterium Pseudomonas hydrogenovora was induced specifically in autotrophic
condition. Dot blot and primer extension analysis showed that the transcription
of the hydrogenase gene started from the region upstream of a hydrogenase
structural gene, hupS, which contained three putative 54-type promoter
sequences (Phup1, Phup2, and Phup3). The lacZ-fusion analysis of the hupS-upstream
region combined with site-directed mutagenesis showed that Phup1 and Phup3
would be essential for a transcriptional initiation. It was also found
that the region upstream of Phup sequences was concerned with regulation
of transcription.
Key words: chemolithoautotroph; gene expression; membrane-bound hydrogenase;
Pseudomonas hydrogenovora; transcriptional regulation
-7-
Synthesis of Didocosahexaenoylphosphatidylserine
Ryuichi Nakashima, Kouichi Kado, Takeshi Kume, and Kazuyuki Maekawa
Osaka Laboratory, Chemicals Inspection and Testing Institute, Japan, 1-6-5 Dogashiba, Tennouji-ku, Osaka 543, Japan
Received May 1, 1997
@1,2-Di-O-isopropylideneglycerophosphorochloridate prepared from isopropylindene
glycerol and phosphorus oxychloride, was allowed to react with Z-L-serine-N-phthalimidomethyl
ester to obtain a derivative of phosphatidylserine. Then, after the isopropylidene
group was removed by Amberlite IR-120 (H+), and the phosphate group was
also blocked as a Ba-salt, this derivative was coupled with docosahexaenoic
acid, applying the method of activated ester. Removal of both protective
groups of serine was finally done by dry hydrogen chloride in chloroform.
Key words: docosahexaenoic acid; phosphatidylserine; didocosahexaenoylphosphatidylserine
-8-
cDNA Cloning and Gene Expression of Phenylalanine Ammonia-Lyase
in Lithospermum erythrorhizon
Kazufumi Yazaki,, Mieko Kataoka, Gisho Honda, Klaus Severin,* and Lutz Heide *,
Faculty of Pharmaceutical Sciences, Kyoto University, Yoshida, Kyoto
606-01, Japan
* Institut fur Pharmazeutische Biologie, Albert-Ludwigs-Universitat Freiburg,
Schanzlestrasse 1, 79104 Freiburg im Breisgau, Germany
Received May 9, 1997
@Two cDNA clones (LEPAL-1 and LEPAL-2) encoding phenylalanine ammonia-lyase
were isolated from cell suspension cultures of Lithospermum erythrorhizon.
Northern kinetic studies showed that LEPAL-1 mRNA contents markedly increased
one day after inoculation of the cells into fresh medium, then decreased
to the steady-state level. The course of mRNA accumulation paralleled that
of PAL enzyme activity. The rapid induction of PAL activity seems to reflect
the induction of dihydroechinofuran biosynthesis, while shikonin was produced
at the steady-state level of PAL activity. The course of LEPAL-2 mRNA accumulation
seemed to be similar to, but much lower than that of LEPAL-1. In the intact
plant, both genes are expressed mainly in the root, the organ in which
shikonin is exclusively produced and accumulated. Genomic Southern blot
analyses showed that both genes are present in the genome as single copies.
Key words: biosynthesis; Lithospermum erythrorhizon; phenylalanine ammonia-lyase;
secondary metabolism; shikonin
-9-
Amino Acid Sequence and Stereoselective Hydrolytic Reaction
of an Endo-1,4--glucanase from a Bacillus Strain
Jun Hitomi, Yuji Hatada, Shunro Kawaminami, Shuji Kawai, and Susumu Ito
Tochigi Research Laboratories of Kao Corporation, 2606 Akabane, Ichikai, Haga, Tochigi 321-34, Japan
Received May 12, 1997
@Bacillus sp. KSM-522 produces three different extracellular endo-1,4--glucanases
[EGs; Okoshi et al., Agric. Biol. Chem., 54, 83-89 (1990)]. Here, we report
the molecular cloning and sequencing of the gene for the fourth EG (EG-IV)
of the organism and the mechanism of its hydrolytic reaction. The structural
gene contained an open reading frame of 1911 bp, corresponding to 636 amino
acids, the amino acid sequence of which was very close to that of an EG
of Clostridium cellulovorans, belonging to the cellulase family E2. The
molecular mass of the extracellular mature enzyme (Ser26 through Lys636)
was calculated to be 69,076 Da, a value close to the 69.2 kDa measured
for the recombinant EG-IV expressed in Bacillus subtilis. The optimum pH
and temperature for activity of the recombinant enzyme were pH 8.0 and
50C, respectively. By 1H-NMR spectroscopy, we demonstrated that the hydrolysis
of p-nitrophenyl -D-cellotrioside by EG-IV proceeded with inversion of
the anomeric configuration.
Key words: Bacillus; cellulase; endoglucanase; family E; inversion
-10-
Transxylosylation of -Xylosidase from Aspergillus awamori
K4
Masahiro Kurakake, Shinji Osada, and Toshiaki Komaki
Department of Food Science & Technology, Fukuyama University, Sanzou, Gakuenchou 1 banchi, Fukuyama, Hiroshima 729-02, Japan
Received May 19, 1997
@-Xylosidase from Aspergillus awamori K4 was purified. The optimum pH
and temperature were around pH 4 and 70C, and the molecular weight was
estimated to be 117,000 on SDS-PAGE analysis. The enzyme has broad acceptor
specificity in transxylosylation. Especially, its acceptor accessibility
for sorbitol and mannitol of sugar alcohols were higher than that for monosaccharides.
Trehalose was a much more effective acceptor than maltose and lactose of
other disaccharides. In the reaction with 13-14% xylooligosaccharides (consisting
of 3.4% xylose, 67.9% xylobiose, and 28.7% xylotriose) and 9-13% acceptors
(sorbitol, mannitol, and trehalose), the amount of transfer products for
each acceptor was 7-11% in 24 h. On 1H- and 13C-NMR analysis, main transfer
products with sorbitol and mannitol were 6-O--xylosyl sorbitol (77.3%)
and 1(6)-O--xylosyl mannitol (73.7%), respectively. Two products with
trehalose were 6(6')-O--xylosyl trehalose (52.1%) and 6,6'-O--di-xylosyl
trehalose (47.9%).
Key words: -xylosidase; transxylosylation; xylooligosaccharide; sugar
alcohol; trehalose
-11-
Effects of a Hydrogenated Isomaltooligosaccharide Mixture
on Glucan Synthesis and on Caries Development in Rats
Jun Tsunehiro, Takashi Matsukubo,* Masao Shiota,** and Yoshinori Takaesu *
Research and Development Center, Showa Sangyo Co., Ltd., Higashi Fukashiba,
Kamisu, Kashima-gun, Ibaraki 314-02, Japan
* Department of Hygiene and Community Dentistry, Tokyo Dental College,
Masago, Mihama-ku, Chiba 261, Japan
** Technical Division, Showa Sangyo Co., Ltd., Higashi Fukashiba, Kamisu,
Kashima-gun, Ibaraki 314-02, Japan
Received May 21, 1997
@The caries inhibitory effect of the hydrogenated derivative of an isomaltooligosaccharides
mixture (IMO-H) was examined in vitro and in vivo experiments. IMO-H could
not be used as a substrate for the crude glucosyltransferases (GTases)
of Streptococcus sobrinus 6715 to synthesize water-insoluble glucan. Moreover,
it not only significantly inhibited the synthesis of water-insoluble glucan
from sucrose, but also the sucrose-dependent adherence of these growing
cells the glass surfaces. In the in vivo experiment, the addition of IMO-H
to a sucrose-containing diet resulted in significant reduction of caries
development in specific-pathogen-free (SPF) rats infected with S. sobrinus
6715.
Key words: caries prevention; hydrogenated isomaltooligosaccharide; sugar
substitute; mutans streptococci; glucan
-12-
Chiral Synthesis of the BC-Ring System of Ciguatoxin from
D-Glucose
Eiichi Ami, Hisakazu Kishimoto,* Hiroshi Ohrui, and Hiroshi Meguro
Department of Applied Biological Chemistry, Faculty of Agriculture,
Tohoku University, Tsutsumidori-Amamiyamachi 1-1, Aobaku, Sendai 981, Japan
* Sumitomo Pharmaceutical Company, Kasugadenaka 3-1-98 Konohana-ku, Osaka
554, Japan
Received May 22, 1997
@The BC-ring system of ciguatoxin was stereoselectively synthesized by
12 steps from methyl -D-glucopyranoside.
Key words: ciguatoxin; chiral synthesis; d-glucose
-13-
New Physiological Effects of 5-Aminolevulinic Acid in
Plants: The Increase of Photosynthesis, Chlorophyll Content, and Plant
Growth
Yasushi Hotta, Tohru Tanaka, Hideo Takaoka, Yasutomo Takeuchi,* and Makoto Konnai *
COSMO Research Institute, 1134-2 Gongendo, Satte, Saitama 340-01, Japan
* Weed Science Center, Utsunomiya University, 350 Minemachi, Utsunomiya,
Tochigi 321, Japan
Received May 23, 1997
@5-Aminolevulinic acid (ALA) promoted the growth and yield of several
crops and vegetables at concentrations lower than those eliciting herbicidal
responses, i.e., less than 1.8 mM by foliar spray and 60 M by root soaking.
To evaluate the physiological action of ALA, the effects of ALA on plants
were examined by several bioassay systems at 0.0006-600 M. ALA at 0.06-6M
by root soaking increased the growth of rice seedlings in light, but did
not affect this in darkness. In horseradish shoot primordia, promotion
by ALA was not proportional among total chlorophyll content, chlorophyll
concentration, and fresh weight. In the test using pothos, ALA at 0.06
M elicited the accumulation of chlorophyll, but the photosynthesis of
the plants was promoted by treatment together with ALA and nutrients. These
results suggest that ALA have a variety of plant physiological effects
on chlorophyll synthesis, photosynthesis, and plant growth, and ALA acts
as a growth regulator in plants at low concentrations. These effects of
ALA were also assumed to be linked to light irradiation and an uptake of
fertilizer by plants. However, excess ALA suppressed these effects.
Key words: 5-aminolevulinic acid; plant growth; chlorophyll; photosynthesis
-14-
Continuous Production of L-Alanine with NADH Regeneration
by a Nanofiltration Membrane Reactor
Shu-Su Lin, Osato Miyawaki, and Kozo Nakamura
Department of Applied Biological Chemistry, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan
Received May 23, 1997
@A conjugated enzyme system, alanine dehydrogenase (AlDH) for stereospecific
reduction of pyruvate to L-alanine and glucose dehydrogenase (GDH) for
regeneration of NADH, were coimmobilized in a nanofiltration membrane bioreactor
(NFMBR) for the continuous production of L-alanine from pyruvate with NADH
regeneration. Since pyruvate was proved to be unstable at neutral pH, it
was kept under acidic conditions and supplied to NFMBR separately from
the other substrates. As 0.2 M pyruvate in HCl solution (pH 4), 10 mM NAD,
0.2 M glucose, and 0.2 M NH4Cl in 0.5 M Tris buffer (pH 8) were continuously
supplied to NFMBR with immobilized AlDH (100 U/ml) and GDH (140 U/ml) at
the retention time of 80 min, the maximum conversion, reactor productivity,
and NAD regeneration number were 100%, 320 g/liter/d, and 20,000, respectively.
To avoid the effect of pyruvate instability, a consecutive reaction system,
lactate dehydrogenase (L-LDH) and AlDH, was also used. In this system,
the L-LDH provides pyruvate, the substrate for the AlDH reaction, from
L-lactate regenerating NADH simultaneously, so the pyruvate could be consumed
as soon as it was produced. As 0.2 M L-lactate, 10 mM NAD, 0.2 M NH4Cl
in 0.5 M Tris buffer (pH 8) were continuously supplied to NFMBR with immobilized
L-LDH (100 U/ml) and AlDH (100 U/ml) at the retention time of 160 min,
the maximum conversion, reactor productivity, and the NAD regeneration
number were 100%, 160 g/liter/d, and 20,000, respectively.
Key words: nanofiltration membrane bioreactor; alanine dehydrogenase; l-lactate
dehydrogenase; glucose dehydrogenase; NADH regeneration
-15-
Superoxide Dismutase Inhibition of Oxidation of Ubiquinol
and Concomitant Formation of Hydrogen Peroxide
Tsutomu Nakayama, Miku Hashimoto, and Kei Hashimoto
School of Food and Nutritional Sciences, University of Shizuoka 52-1 Yada, Shizuoka 422, Japan
Received May 27, 1997
@We measured ubiquinone (CoQ0) and hydrogen peroxide (H2O2) formed in
the process of oxidation of ubiquinol (CoQ0H2). We found that copper-zinc
superoxide dismutase and manganese superoxide dismutase inhibited both
the CoQ0 formation and the H2O2 formation only in the presence of chelators
such as DTPA (diethylenetriaminepentaacetic acid). The amount of H2O2 was
almost equal to that of CoQ0, indicating that the H2O2 formation was coupled
with the CoQ0 formation. The lack of inhibitory effects of the corresponding
heat-inactivated superoxide dismutase (SOD) confirmed that the inhibition
by the original SOD was due to its enzymatic activity. We propose that
CoQ0H2 oxidation occurs as a chain reaction with superoxide as the chain
carrier and that SOD inhibits this reaction by lowering the superoxide
concentration.
Key words: superoxide dismutase (SOD); superoxide; hydrogen peroxide; ubiquinol;
ubiquinone
-16-
Inactivation of Cu,Zn-Superoxide Dismutase by Intermediates
of Maillard Reaction and Glycolytic Pathway and Some Sugars
Hiroyuki Ukeda, Yuko Hasegawa, Toshinao Ishi, and Masayoshi Sawamura
Department of Bioresources Science, Faculty of Agriculture, Kochi University, Monobe B-200, Nankoku 783, Japan
Received May 30, 1997
@Human Cu,Zn-superoxide dismutase (SOD) was incubated with various intermediates
of the Maillard reaction and glycolytic pathway (arabinose, glyoxal, glycolaldehyde,
glyceraldehyde, glyceraldehyde 3-phosphate, and dihydroxyacetone) and some
reducing sugars (sorbose, xylose, and ribose). The change of the activity
and the molecular weight were measured and compared with that of SOD incubated
with glucose or fructose. Sorbose, xylose, and ribose decreased the activity
with a rate comparable to fructose. Site-specific and random fragmentation
were observed upon the incubation with them. Arabinose showed a similar
inactivation rate as glucose. The intermediates other than arabinose had
a high inactivation rate. Especially, glyceraldehyde, glycolaldehyde, and
glyoxal most strongly lowered the activity in a concentration-dependent
manner and a significant inactivation was recognized even at 1 mM level.
SDS-PAGE band patterns indicated that the inactivation by those carbonyl
compounds occurred by both crosslinking and site-specific fragmentation
of SOD.
Key words: superoxide dismutase; glycation; Maillard reaction; inactivation;
glycolytic pathway
-17-
Synthesis and Biological Activities of 8'-Methylene- and
8'-Methylidyneabscisic Acids
Yasushi Todoroki, Sei-ichi Nakano, Sho Arai, Nobuhiro Hirai, and Hajime Ohigashi
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwake-cho, Sakyo-ku, Kyoto 606-01, Japan
Received May 30, 1997
@8'-Methylene- and 8'-methylidyneabscisic acids which might act as suicide
inhibitors of the 8'-hydroxylase of abscisic acid, were designed, synthesized,
and optically resolved. The (+)-isomers showed stronger inhibitory activity
in rice elongation and in lettuce seed germination than (+)-abscisic acid.
The activity of (+)-8'-methylidyneabscisic acid was the strongest of the
analogues synthesized to date, 40-fold stronger than abscisic acid.
Key words: abscisic acid; 8'-methyleneabscisic acid; 8'-methylidyneabscisic
acid; cytochrome P-450 monooxygenase; suicide inhibitor
-18-
Purification and Characterization of N-Acetylneuraminate
Synthase from Escherichia coli K1-M12
Eriko Komaki, Yasuhiro Ohta, and Yoji Tsukada
Kyoto Research Laboratories, Marukin Shoyu Co., Ltd., 27 Monnomae, Todo, Uji-shi, Kyoto 611, Japan
Received May 30, 1997
@N-Acetylneuraminate (NeuAc) synthase, which catalyzes NeuAc synthesis
by condensation of N-acetyl-D-mannosamine (ManNAc) and phosphoenolpyruvate
(PEP), was purified from a cell extract of Escherichia coli K1-M12 to electrophoretically
homogeneity by serial column chromatographies. The molecular weight of
native enzyme was estimated to be 106,000 by gel filtration. After denaturation
in sodium dodecyl sulfate, the molecular weight was reduced to 52,000,
indicating the existence of 2 identical subunits. The optimum pH was 7.5
and the stable pH range was 7.0 to 10.0. The enzyme was thermostable up
to 30C. No metal ion was required for the enzyme activity. SH-inhibitors
such as p-chloromercuribenzoic acid and mercury chloride were potent inhibitors.
The Km for ManNAc and PEP were 5.6 mM and 0.04 mM, respectively.
Key words: N-acetylneuraminate synthase; N-acetyl-d-mannosamine; N-acetylneuraminate
-19-
Modulation of GABA Receptors Expressed in Xenopus Oocytes
by 13-L-Hydoxylinoleic Acid and Food Additives
Hitoshi Aoshima and Yukihiro Tenpaku
Department of Biology, Physics and Informatics, Faculty of Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753, Japan
Received June 2, 1997
@To study the effects of 13-L-hydroxylinoleic acid (LOH) and food additives
on -aminobutyric acid (GABA) receptors, ionotropic GABA receptors were
expressed in Xenopus oocytes by injecting mRNAs prepared from rat whole
brain. LOH, which was prepared by reduction of 13-L-hydroperoxylinoleic
acid (LOOH), inhibited the response of GABA receptors in the presence of
high concentrations of GABA. LOH also inhibited nicotinic acetylcholine,
glycine, and kainate receptors, while it had little effect on NMDA receptors
expressed in Xenopus oocytes. However, LOH potentiated the response of
GABA receptors as well as LOOH in the presence of low concentrations of
GABA, possibly increasing the affinity of GABA for the receptors, while
linoleic acid did not. Since some modification of the compounds seemed
to change their effects on GABA receptors, the responses of GABA receptors
elicited by 10 M GABA were measured in the presence of compounds with
various kinds of functional groups or the structural isomers of pentanol.
Potentiation of GABA receptors depended strongly on the species of functional
groups and also depended on the structure of the isomers. Then effects
of various kinds of food additives on GABA receptors were also examined;
perfumes such as alcohols or esters potentiated the responses strongly,
while hexylamine, nicotinamide, or caffeine inhibited the responses, mainly
in a competitive manner, and vanillin inhibited the responses noncompetitively.
These results suggest the possibility that production of LOOH and LOH,
or intake of much of some food additives, modulates the neural transmission
in the brain, especially through ionotropic GABA receptors and changes
the frame of the human mind, as alcohol or tobacco does.
Key words: food additive; GABA receptor; hydroxylinoleic acid; potentiation;
Xenopus oocyte
-20-
Characterization of Styrene Oxide Isomerase, a Key Enzyme
of Styrene and Styrene Oxide Metabolism in Corynebacterium sp.
Nobuya Itoh,*, Kunimasa Hayashi, Keisaku Okada, Takeshi Ito, and Naoyuki Mizuguchi
Department of Applied Chemistry and Biotechnology, Faculty of Engineering,
Fukui University, Bunkyo 3-9-1, Fukui 910, Japan
* Biotechnology Research Center, Toyama Prefectural University, 5180 Kurokawa
Kosugi, Toyama 939-03, Japan
Received June 5, 1997
@Styrene oxide isomerase (SOI) [EC 5.3.99.7], most probably located in
the cell wall, was partially purified from Corynebacterium sp. AC-5 cells
grown in a styrene gas atmospheres. The enzyme catalyzed the isomerization
reaction to give phenylacetaldehyde, but did not catalyze its reverse reaction.
The optimum pH of the reaction was around 7.0, and the enzyme was unstable
below pH 6.0. The Km toward styrene oxide was very low (7.7~10-5 M), indicating
its high affinity for styrene oxide. The enzyme showed strict substrate
specificity, and epoxide compounds other than styrene oxide did not serve
as substrates. (S )-Styrene oxide was preferentially converted by the enzyme,
compared with the (R)-isomer. The possible application of SOI as a biocatalyst
is also discussed.
Key words: styrene oxide isomerase; styrene; styrene oxide; metabolism;
Corynebacterium sp.
-21-
Glycogen-Surfactant Complexes: Phase Behavior in a Water/Phytoglycogen/Sodium
Dodecyl Sulfate (SDS) System
Akihiro Nakano, Ryozo Irie,* and Koichi Tateishi *
The United Graduate School of Agricultural Science, Gifu University
(Shinshu University), Yanagido 1-1, Gifu 501-11, Japan
* Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu
University, Minamiminowa 8304, Nagano 399-45, Japan
Received June 9, 1997
@The phase behavior was investigated in water/phytoglycogen/SDS and C12EO20
systems. It was found that phytoglycogen was precipitated in the presence
of these surfactants. Phytoglycogen in the SDS system was dispersed more
homogeneously than in the C12EO20 system, and a liquid-liquid phase separation
region appeared in the SDS system. In the C12EO20 system, the transmittance
of a stirred solution in the single-phase region was slightly lower than
that of the phytoglycogen controls, whereas the transmittance of the stirred
solution was higher than that of the controls in the SDS system. It thus
seems that phytoglycogen formed a complex with SDS and that this complex
affected the transmittance in the single-phase region. Viscosity measurements
of the aqueous solution supported the existence of such a phytoglycogen-SDS
complex. These results enabled us to propose a schematic representation
of the complex structure. It was clarified that phytoglycogen forms a complex
with SDS, and that the structure of the complex modifies the dispersion
stability of phytoglycogen.
Key words: glycogen-surfactant complex; starch-surfactant complex; phytoglycogen;
ionic surfactant; phase behavior
-22-
Autoxidation Reaction Mechanism for L-Ascorbic Acid in
Methanol without Metal Ion Catalysis
Noriko Miyake, Yuzuru Otsuka,* and Tadao Kurata
Institute of Environmental Science for Human Life, Ochanomizu University,
Bunkyo-ku, Tokyo 112, Japan
* Faculty of Education, Tottori University, Tottori-shi, Tottori 680, Japan
Received June 10, 1997
@The autoxidation reaction of L-ascorbic acid (ASA) in methanol without
metal ion catalysis was studied. Besides L-threonolactone (THL) and oxalic
acid (OXA), methyl L-threonate, and threonic acid were identified as initial
autoxidation products of ASA, which were the C(2)-C(3) fission product
via the C(2) oxygen adduct of ASA. This pathway is different from the one
via dehydro-L-ASA (DASA), which has long been believed to be the only oxidation
pathway of ASA. It was confirmed that this reaction also occurred in both
water and other polar solvents, including methanol. It was clarified that
mono-dissociated ASA was more reactive than the non-dissociated ASA in
this pathway, and that the main reaction products formed from these two
forms of ASA were also somewhat different. Determination of the amount
of remaining ASA and the yields of THL and OXA, C(2)-C(3) fission products,
and of DASA were carried out doing the autoxidation of ASA under various
reaction conditions.
Key words: l-ascorbic acid; autoxidation; l-ascorbate; l-threonic acid;
oxygen adduct of ascorbic acid
-23-
Molecular Cloning of Levan Fructotransferase Gene from
Arthrobacter nicotinovorans GS-9 and Its Expression in Escherichia coli
Katsuichi Saito, Atsushi Yokota, and Fusao Tomita
Laboratory of Applied Microbiology, Department of Bioscience and Chemistry, Faculty of Agriculture, Hokkaido University, Sapporo 060, Japan
Received June 13, 1997
@The gene encoding an extracellular levan fructotransferase, designated
the lft gene, was cloned from the genomic DNA of Arthrobacter nicotinovorans
GS-9, and expressed in Escherichia coli. It was found that a single open
reading frame consisted of 1554 base pairs that encoded a polypeptide composed
of a signal peptide of 33 amino acids and a mature protein of 484 amino
acids (Mr 53,152), and it was also found that a putative ribosome-binding
site was present in the upstream from the ORF.
@The primary structure had no significant similarity with those of inulin
fructotransferases, but had low similarity to the catalytic regions of
other fructosylhydrolases.
@The expression of the lft gene was increased on a plasmid, pLFT-BB1,
in which the lft gene was fused with -peptide of the lacZ gene of pUC18.
An E. coli transformant carrying pLFT-BB1 expressed six times as much activity
of levan fructotransferase as that of the original strain, A. nicotinovorans
GS-9.
Key words: levan; levan fructotransferase; DFA IV; lft gene; Arthrobacter
nicotinovorans GS-9
-24-
Effect of Polyphenol Oxidase on Deodorization
Osamu Negishi and Tetsuo Ozawa
Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305, Japan
Received June 13, 1997
@A mixture of purified polyphenol oxidases (PPO), or acetone powders prepared
from fruits and vegetables, and polyphenolic compounds (PPs) totally eliminated
a methylmercaptan odor. 2-Methylthiochlorogenic acid was isolated from
the reaction mixture of methylmercaptan and chlorogenic acid with burdock
acetone powder. Further, the formation of 5-methylthiochlorogenic acid
and 2,5-bis(methylthio)chlorogenic acid was suggested. These facts demonstrate
that the o-quinone compounds formed from o-diphenols by PPO rapidly reacted
with methylmercaptan. The oxidation reaction of PPs by using acetone powder
containing PPO or peroxidase is considered to be more effective for removing
bad smells from our mouths and from the environment.
Key words: deodorization; polyphenol oxidase; polyphenolic compound; peroxidase;
2-methylthiochlorogenic acid
-25-
Oxidative Stability of Docosahexaenoic Acid-containing
Oils in the Form of Phospholipids, Triacylglycerols, and Ethyl Esters
Jin-Hyang Song, Yoshikazu Inoue,* and Teruo Miyazawa
Department of Applied Biological Chemistry, Faculty of Agriculture,
Tohoku University, Sendai 981, Japan
* Bizen Chemical Co., Tokutomi, Okayama 709-07, Japan
Received June 27, 1997
@The peroxidative stability of docosahexaenoic acid (DHA)-containing oils
(DHA at 10.7 mol% of the total fatty acids), in the form of phospholipids
(PL), triacylglycerols (TG), and ethyl esters (EE) with the same constituent
fatty acids, was investigated in the dark at 25C in a bulk phase, and
compared with that of control palm oil (supplemented with 20% soybean oil).
The oxygen absorption of the DHA-containing oil was significantly lower
in the form of PL than in the form of TG and EE during a 10-week oxidation,
and the oxygen absorption of PL was almost equivalent to that of the control
oil. A gas chromatographic analysis showed that 90% of initial DHA was
retained in the form of PL after the 10-week oxidation, while TG and EE
respectively more rapidly decayed with the loss of 97% and 64% of DHA.
Tocopherol in the form of TG and EE had also completely decayed after the
oxidation, while 37% of the initial tocopherol remained in the form of
PL. The peroxide and carbonyl values of TG and EE showed large increases
after the oxidation, but no such increase was observed for PL. These results
show that DHA-containing oil in the form of PL was more resistant to the
oxidative degradation of DHA than that in the form of TG and EE in a bulk
phase.
Key words: oxidative stability; docosahexaenoic acid; oxygen absorption;
tocopherol; phospholipid
-26-
Effects of Dietary Lipid Peroxidation Products on Hormonal
Responses in Primary Cultured Hepatocytes of Rats
Hitoshi Ashida, Kenichi Ohue, Kazuki Kanazawa, and Gen-ichi Danno
Department of Biofunctional Chemistry, Faculty of Agriculture, Kobe University, Rokkodai-cho, Nada-ku, Kobe 657, Japan
Received July 2, 1997
@Dietary lipid peroxidation products cause endogenous lipid peroxidation
with hepatic dysfunction. In this study, we isolated and cultured hepatocytes
of rats that were given secondary autoxidation products of linoleic acid
(p.o., 400 mg/rat/day for 3 days), and examined the hormonal responses
of these hepatocytes. An increase in thiobarbituric acid reactive substances
and a depletion of vitamin E persisted in hepatocytes from treated rats
for at least 24 h in culture as compared to those from control rats. As
markers for hepatic dysfunction, the activities of six enzymes were measured.
In each case, there was an initial decrease in the enzyme activity in hepatocytes
from the treated rats, and all activities were restored by 48 h in culture.
Then, we measured the hormonal responses of these hepatocytes. The responses
to insulin or glucagon in hepatocytes from secondary products-treated and
control rats were the same. In contrast, the response to dexamethasone
was significantly lowered in hepatocytes from secondary products-treated
rats as measured by the induction of tryptophan 2,3-dioxygenase and tyrosine
aminotransferase. We conclude that primary cultured hepatocytes from the
rats treated in vivo with dietary lipid peroxidation products retained
symptoms of oxidative stress and had a low response to glucocorticoids.
Key words: lipid peroxidation; glucocorticoid; primary cultured hepatocytes;
rats
-27-
Identification of the Essential Amino Acid Residues in
LukS for the Hemolytic Activity of Staphylococcal Leukocidin towards Rabbit
Erythrocytes
Hirofumi Nariya, Akiko Shimatani, Toshio Tomita, and Yoshiyuki Kamio
Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku University, 1-1 Amamiya-machi, Tsutsumi-dori, Aoba-ku, Sendai 981, Japan
Received July 10, 1997
@Staphylococcus aureus V8 (ATCC 49775) produces Panton-Valentine leukocidin
(PVL) and leukocidin (Luk). PVL and Luk consist of LukF-PV and LukS-PV,
and LukF and LukS, respectively. LukF and LukS cooperatively and strongly
lyse rabbit erythrocytes besides human and rabbit polymorphonuclear leukocytes.
LukF and LukS-PV also cooperatively lysed rabbit erythrocytes, but its
activity was only 4% of that in combination [LukF-LukS] after 15 min of
incubation. To identify the pivotal region responsible for the hemolytic
function of LukS towards rabbit erythrocytes, we created a series of chimeric
genes (lukS-PV/lukS ) and mutant genes of lukS-PV and had them expressed
in Escherichia coli. The chimeric and mutant proteins purified from the
sonicated extract from the cells of E. coli were assayed for their hemolytic
activities towards rabbit erythrocytes in combination with LukF or LukF-PV.
The results indicate that a 2-residue segment (D12I13) of lukS is the minimum
region essential for the hemolytic function of LukS towards rabbit erythrocytes.
Key words: Staphylococcal leukocidin; Panton-Valentine leukocidin; LukF-PV;
LukS-PV
-28-
Note
Purification and Some Properties of a Protease from the Sarcocarp of Musk
Melon Fruit
Makoto Kaneda, Hiroo Yonezawa, and Tetsuya Uchikoba
Department of Chemistry, Faculty of Science, Kagoshima University, Korimoto 1-21-35, Kagoshima 890, Japan
Received April 8, 1997
@A protease has been purified from sarcocarp of musk melon, Cucumis melo
ssp. melo var. reticulatus Naud. Earl's Favourite. The protease was mostly
present in the placenta part of the fruit and next in the inside mesocarp.
The molecular mass of the enzyme was estimated to be about 62 kDa on SDS-PAGE.
The enzyme had a carbohydrate moiety. The optimum pH of the enzyme was
11 at 35C using casein as a substrate. The enzyme was stable between
pH 6 and 11. The enzyme was strongly inhibited by diisopropyl fluorophosphate,
but was not inhibited by EDTA or cysteine protease inhibitors. From the
digestion of Ala-Ala-Pro-X-pNA (X=Phe, Leu, Val, Ala, Gly, Lys, Glu, Pro,
and diaminopropionic acid (Dap) substrates the specificity of the protease
was found to be approximately broad, but the preferential cleavage sites
were C-terminal sites of hydrophobic or acidic amino acid residues at P1
position. It was proved that the enzymatic properties of musk melon protease
are similar to those of cucumisin [EC 3.4.21.25]. The enzyme was not inhibited
by typical proteinous inhibitors such as STI or ovomucoid. Therefore, this
enzyme seems to be a useful protease for the food industries.
Key words: Cucumis melo; Cucurbitaceae; musk melon; plant protease; serine
protease
-29-
Note
Synthesis and Fungicidal Activity of 6-Alkyl Six-membered Cyclic Thiophosphates
Shinkichi Tawata, Shigehiko Taira, Hirofumi Kikizu, Naotada Kobamoto, Masanobu Ishihara, and Seizen Toyama
Department of Bioscience and Biotechnology, College of Agriculture, University of the Ryukyus, 1 Senbaru, Nishihara-cho, Okinawa 903-01, Japan
Received April 16, 1997
@Synthesis and fungicidal activities of new 6-alkyl six-membered cyclic
phosphates were examined. Ten kinds of 6-alkyl six-membered cyclic thiophosphates
were synthesized by reaction with 5-alkyl-2-hydroxybenzyl alcohols and
phosphoric agents. Among the prepared compounds, 2-ethoxy-6-ethyl-4H-1,3,2-benzodioxaphosphorin
2-sulfide (1) had activity as potent as the commercial fungicide iprobenfos
against Pythium sp. and Corticium rolfsii at 10 ppm.
Key words: antifungal activity; 6-ethyl-2-ethoxy-4H-1,3,2-benzodioxaphosphorin
2-sulfide; 6-isopropyl-2-ethoxy-4H-1,3,2-benzodi oxaphosphorin 2-sulfide;
Pythium sp.; Corticium rolfsii
-30-
Note
Superoxide-scavenging and Tyrosinase-inhibitory Activities of the Extracts
of Some Chinese Medicines
Zhuang Miao, Hiroshi Kayahara,* and Koji Tadasa*
The United Graduate School of Agricultural Science, Gifu University,
1-1 Yanagido, Gifushi, Gifu 501-11, Japan
* Division of Bio-organic Chemistry, Department of Bioscience and Biotechnology,
Faculty of Agriculture, Shinshu University, 8304 Minamiminowamura, Kamiinagun,
Nagano 399-45, Japan
Received April 21, 1997
@The superoxide-scavenging and the tyrosinase-inhibitory activities of
28 kinds of plants used as Chinese medicines were evaluated. Methanol/water
extracts were used for the screening tests, and for those which represented
high activities, other kinds of extracts were also studied. The extracts
of Mallotus japonicus Muell. Arg. scavenged superoxide strongly; the half-inhibiting
concentration (IC50) of its 50% methanol/water extract was 10.57 g of
dried material in 1 ml of reaction mixture. The extracts of Fritillaria
thunbergii Miq., Carthamus tinctorius L., and Prunus persica (L.) Batsch
had strong tyrosinase-inhibitory activities, and the extracts of Scutellaria
baicalensis Georgi represented both kinds of activities. These facts suggested
that Chinese medicines may be a treasure house of chemical compounds that
have the superoxide-scavenging and the tyrosinase-inhibitory activities.
Key words: Chinese medicine; superoxide-scavenging activity; tyrosinase-inhibitory
activity
-31-
Note
Formation of Thioxopyrrolidines and Dithiocarbamates from 4-Methylthio-3-butenyl
Isothiocyanates, the Pungent Principle of Radish, in Aqueous Media
Hiroki Matsuoka, Yoshinori Toda, Kenji Yanagi, Asaka Takahashi, Koichi Yoneyama,* and Yasushi Uda
Laboratory of Food Chemistry, Department of Bioproductive Sciences,
and
* Weed Science Center, Utsunomiya University, 350 Mine, Utsunomiya 321,
Japan
Received May 15, 1997
@Reaction products of 4-methylthio-3-butenyl isothiocyanate (MTBI), the
radish pungent principle, in aqueous media were identified and their antimicrobial
activities were examined. A rapid degradation of MTBI in aqueous media
afforded a mixture of 3-(hydroxy)methylene-2-thioxopyrrolidine (1), (Z)-3-(methylthio)methylene-2-thioxopyrrolidine
(2), its (E)-isomer (3), methyl 4-methylthiobutyldithiocarbamate (4), methyl
(Z)-4-methylthio-3-butenyldithiocarbamate (5), and its (E)-isomer (6).
The products 1, 2, and 3 were detected at all pHs examined, while 4, 5,
and 6 were formed at pH over 6.0. The formation of 4 from 6 was accompanied
by an oxidation of methanethiol released from MTBI in aqueous media. Antimicrobial
activities of 2 and 3 against all microbes examined were much lower than
that of 1, which had MICs ranging from 50 to 400 g/ml. As for 4, 5, and
6, antifungal activities were comparable to that of 1, but little antibacterial
activities were observed. The antimicrobial activities of the six products
were considered to be far lower than that of MTBI.
Key words: radish pungent principle; thioxopyrrolidines; dithiocarbamates;
antimicrobial activity
-32-
Note
Diurnal Fluctuation in the Enzyme Activity and the Messenger RNA Level
of Pineal Serotonin N-Acetyltransferase in Normal and Hereditary Microphthalmic
Rats
Suhyeun Shim,* Zhengwei Fu,* Hisanori Kato, and Hideyuki Tanaka
Departments of Applied Biological Chemistry and Animal Sciences, Faculty
of Agriculture, Utsunomiya University, Mine-350, Utsunomiya, Tochigi 321,
Japan
* Department of Applied Biochemistry, United Graduate School, Tokyo University
of Agriculture and Technology, Fuchu, Tokyo 183, Japan
Received May 19, 1997
@The enzyme activity and the messenger RNA level of pineal serotonin N-acetyltransferase
were more than 20- and 50-fold higher, respectively, in the dark period
than in the light period in normal rats. In hereditary microphthalmic rats,
however, the serotonin N-acetyltransferase activity and its mRNA level
did not undergo a great diurnal change through the light and dark periods.
These results indicate that the diurnal rhythms of the activity and the
mRNA level of serotonin N-acetyltransferase are not detected in the pineal
gland of hereditary blind rats, suggesting free-running rhythms in individual
animals due to desynchronization of their circadian rhythms by a lack of
their optic nerve.
Key words: circadian rhythm; serotonin N-acetyltransferase mRNA; hereditary
blind rat; pineal
-33-
Note
The Effects of Niacin on DNA Repair after N-Methyl-N'-nitro-N-nitrosoguanidine
Treatment in Normal Human Lymphocytes
Shin Ogata, Katsuzumi Okumura, and Hiroshi Taguchi
Laboratory of Biological Chemistry, Faculty of Bioresources, Mie University, Tsu, Mie 514, Japan
Received May 30, 1997
@We have investigated the effects of niacin on NAD levels and on DNA repair
in human lymphocytes. When lymphocytes were incubated in culture medium
with various concentrations of niacin, incubation of lymphocytes with nicotinic
acid at 5 M or nicotinamide at 10 mM caused a 2-3 fold increase in NAD
content. Under these conditions lymphocytes were treated with N-methyl-N'-nitro-N-nitrosoguanidine
(MNNG). Interestingly, the rejoining of DNA strand breaks was promoted
by nicotinic acid but nicotinamide inhibited the rejoining.
Key words: niacin; DNA repair; human lymphocytes; poly(ADP-ribose)polymerase;
N-methyl-N'-nitro-N-nitrosoguanidine
-34-
Note
Regulatory Region of Expression of Thiobacillus ferrooxidans leuB Gene
in Escherichia coli
Hiroshi Kawaguchi, Kenji Inagaki, and Hidehiko Tanaka
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Okayama 700, Japan
Received June 2, 1997
@The regulatory region of Thiobacillus ferrooxidans leuB gene was analyzed.
The deletion analysis indicated that the promoter sequences CATCCG and
TATTAT, which were similar to the consensus -35 and -10 sequences of Escherichia
coli, respectively, existed. The transcriptional initiation site was located
at the position of cytosine -26. The deletion analysis of the upstream
region suggested the existence of a regulatory region by leucine and the
region related to transcription of the gene.
Key words: Thiobacillus ferrooxidans; 3-isopropylmalate dehydrogenase;
leuB; gene expression; regulatory mechanism
-35-
Note
Starvation-increased Insulin-dependent Tyrosine Phosphorylation of the
195-kDa Protein in Intact Rat Liver
Yoshiaki Ito, Shin-Ichiro Takahashi,* Asako Takenaka, Tomomi Hidaka, and Tadashi Noguchi
Department of Applied Biological Chemistry, and * Department of Applied Animal Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113, Japan
Received June 12, 1997
@Insulin stimulates tyrosine phosphorylation of 175-195 kDa proteins including
insulin receptor substrate-1 (IRS-1) in various tissues and cell types.
In intact rat livers, starvation increased the insulin-dependent tyrosine
phosphorylation of the insulin receptor and IRS-1 as has been described
by others. Surprisingly, starvation greatly increased the tyrosine phosphorylation
of the 195-kDa protein induced by insulin, indicating that this protein
may be a new substrate of the insulin receptor kinase. The marked increase
in tyrosine phosphorylation of the 195-kDa protein may have a physiological
role in signal transmission in response to insulin under starvation conditions.
Key words: insulin; tyrosine phosphorylation; insulin receptor; insulin
receptor substrate-1 (IRS-1); rat liver
-36-
Note
Periplasmic Secretion of Functional Ovotransferrin N-Lobe in Escherichia
coli
Tsuguo Miyauchi, Nobuyuki Takahashi, and Masaaki Hirose
The Research Institute for Food Science, Kyoto University, Uji, Kyoto 611, Japan
Received June 13, 1997
@The cytoplasmic and periplasmic production systems of ovotransferrin
N-lobe in Escherichia coli were constructed. The periplasmic, but not cytoplasmic
product, was found to have the iron-binding function.
Key words: ovotransferrin; periplasmic secretion; egg white protein; recombinant
ovotransferrin; iron-binding protein
-37-
Note
Eicosanyl p-Coumarates from a Kenyan Plant, Psiadia punctulata : Plant
Growth Inhibitors
Joseph M. Keriko,* Shuhei Nakajima,** Naomichi Baba,** and Junkichi Iwasa***
*Graduate School of Natural Science and Technology, Okayama University,
3-1-1 Tsushima-naka, Okayama 700, Japan
** Okayama University, Faculty of Agriculture, 1-1-1 Tsushima-naka, Okayama
700, Japan
*** Jomo Kenyatta University, Faculty of Agriculture, P.O. Box 62000, Nairobi,
Kenya
Received June 16, 1997
@From methanol extracts of fresh leaves of Psiadia punctulata (DC.) Vatke,
lettuce seed radicle growth inhibitors were isolated and identified as
E- and Z-eicosanyl p-coumarates by spectroscopic analyses.
Key words: Psiadia punctulata (DC.) Vatke; Asteraceae; radicle growth inhibition;
lettuce seeds; E- and Z-eicosanyl p-coumarates
-38-
Note
Production of New Restriction Endonuclease VpaK11BI Recognizing Sequence
5'-GGWCC-3' in a Haemolysin-less Mutant of Vibrio parahaemolyticus
Harumi Ueno, Akihiko Kita, Michiko Miyahara,* Naoko Ishiwata,* Katsutoshi Mise,* Ikunoshin Kato, and Yoshizumi Ishino
Biotechnology Research Laboratories, Takara Shuzo Co., Ltd., Seta 3-4-1,
Otsu, Shiga 520-21, Japan
* National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku,
Tokyo 158, Japan
Received June 16, 1997
@High restriction endonuclease activity was found in a haemolysin-less
mutant of the Vibrio parahaemolyticus 1743-1 strain. The endonuclease,
named VpaK11BI, recognized the palindromic pentanucleotide sequence of
5'-GGWCC-3' and cleaved double-stranded DNA after the first G, which is
exactly the same as the specificity of AvaII. The haemolysin-less mutant
of V. parahaemolyticus is now available for producing the valuable restriction
endonuclease on a commercial scale.
Key words: restriction endonuclease; AvaII isoschizomer; Vibrio parahaemolyticus;
haemolysin-less mutant
-39-
Note
Mutational Analysis of the Potato Virus Y 5' Untranslated Region for Alteration
in Translational Enhancement in Tobacco Protoplasts
Li Jun Yang, Makoto Hidaka,* Junichiro Sonoda, Haruhiko Masaki, and Takeshi Uozumi
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan
Received June 16, 1997
@The 185 nucleotide 5' untranslated region (5'UTR) of potato virus Y ordinary
strain (PVY-O) showed translation-enhancing activity on the -glucuronidase
(GUS) gene in tobacco protoplasts. Mutational analysis of the 5'UTR was
done to find sequence motifs necessary for the enhancement. Deletions within
the 1-130 nucleotide region of 5'UTR stimulated the GUS expression in some
cases, while the GUS activity declined with deletions in the 131-185 nucleotide
region. The results indicated that the last 55 nucleotides of PVY-O 5'UTR
might play the much important role in the translational enhancement in
plant cells.
Key words: potato virus Y; 5' untranslated region; translational enhancement;
mutational analysis
-40-
Note
Conversion from Tryptophan Precursor into Violacein Pigments by a Cell-free
System from Chromobacterium violaceum
Tsutomu Hoshino and Masahiro Yamamoto
Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University, Ikarashi, Niigata 950-21, Japan
Received June 20, 1997
@A cell-free system for converting tryptophan precursor into violacein
pigments is reported. Crucial factors were the requirements of the reduced
nicotinamides as a cofactor and Zn2+ as a metal ion. Optimal pHs were in
the range of 8.5-9.5. The effectiveness of NADH was thirty times lower
than that of NADPH at a low concentration of 2 mM. The oxygenation mechanism(s)
is discussed by using oxygenase inhibitors such as amethopterin, metyrapone,
ancymidol, and prohexadione. Metal-chelating agents strongly inhibited
the biosynthesis, suggesting that metallo-enzymes are involved in the biosynthesis.
Key words: tryptophan; violacein; cell-free biosynthesis; NADPH; metallo-enzyme
-41-
Note
Effects of Carnosine and Anserine on the Destruction of Vitamin B(/)12
with Vitamin C in the Presence of Copper
Shigeo Takenaka,* Sumi Sugiyama,* Fumio Watanabe,** Katsuo Abe,** Yoshiyuki Tamura,* and Yoshihisa Nakano***
* Laboratory of Nutrition and Food Science, Hagoromo-gakuen College,
Sakai 592, Japan
** Department of Food and Nutrition, Kochi Women's University, Kochi 780,
Japan
*** Department of Applied Biological Chemistry, Osaka Prefecture University,
Sakai 593, Japan
Received June 23, 1997
@Vitamin B(/)12 is destroyed by the addition of substantial amounts of
vitamin C in the presence of copper. Effects of carnosine and anserine,
natural water-soluble antioxidants, on the destruction of vitamin B(/)12,
were studied. Addition of carnosine (10 mm) effectively repressed the destruction
of vitamin B(/)12, but anserine had only weak inhibitory effects.
Key words: anserine; antioxidants; carnosine; cobalamin; vitamin C
-42-
Note
Purification and Characterization of Two Chitinase Isoforms from the Bulbs
of Gladiolus (Gladiolus gandavensis)
Takeshi Yamagami, Yoichiro Mine, Yoichi Aso, and Masatsune Ishiguro
Laboratory of Protein Chemistry & Engineering, Faculty of Agriculture, Kyushu University, Fukuoka 812-81, Japan
Received June 26, 1997
@Two chitinase isoforms, designated GBC-a and GBC-b, were purified from
the bulbs of gladiolus (Gladiolus gandavensis) using CM-cellulose column
chromatography followed by Butyl-Toyopearl 650M hydrophobic column chromatography,
gel filtration on Sephadex G-75, and Mono-S FPLC. GBC-a and GBC-b are weakly
acidic and weakly basic proteins with molecular masses of 30 kDa, and isoelectric
points of 6.0 and 7.5, respectively. GBC-a and GBC-b were found to be homologous
proteins with similar amino acid compositions and N-terminal sequences.
The number of half-cystine residues in GBC-a and GBC-b was only one each,
which is much lower than those of plant class I (15-17 Cys residues/mol),
class II (5-8 Cys residues/mol), and class III (6 Cys residues/mol) chitinases.
The N-terminal sequences of GBC-a and GBC-b were completely different from
those of plant three classes of chitinases. The optimal pHs of these chitinases
toward glycolchitin were pH 5. GBC-a hydrolyzed (GlcNAc)6 into (GlcNAc)2,
(GlcNAc)3 and (GlcNAc)4, and (GlcNAc)5 into (GlcNAc)2 and (GlcNAc)3.
Key words: chitinase; plant bulb; gladiolus
-43-
Note
Molecular Cloning and Characterization of a cDNA Encoding Early Light-inducible
Protein from Soybean (Glycine max L.)
Hiroshi Yamagata and Chris Bowler*
Laboratory of Biochemistry, Faculty of Agriculture, Kobe University,
Nada, Kobe 657, Japan
* Stazione Zoologica, Villa Comunale, I 80121 Naples, Italy
Received June 27, 1997
@A cDNA for early light-inducible protein (ELIP) was isolated from Glycine
max L. and the nucleotides were sequenced. The cDNA encodes a 192-residue
polypeptide of 20.3 kDa. The deduced amino acid sequence included a transit
peptide in the amino-terminus and three hydrophobic regions long enough
to span the thylakoid membranes, and had a high similarity to those of
ELIPs and related polypeptides from other plants.
Key words: early light-inducible protein (ELIP); cDNA cloning; Glycine
max L.
-44-
Short Communication
Flutolanil Resistance as a Genetic Marker of Coprinus cinereus Strains
Shigeru Oita, Tsutomu Fushimi, Tetsuya Ookura, Yasuhiro Ito, and Sonoe O. Yanagi
National Food Research Institute, Ministry of Agriculture, Forestry, and Fisheries, 2-1-2 Kannondai, Tsukuba 305, Japan
Received May 19, 1997
@All 4 Schizophyllum commune strains among several basidiomycete species
were exclusively resistant to flutolanil, which inhibits succinate dehydrogenase
complex (SDC) function. Flutolanil resistant strains could be screened
from monokaryotic protoplasts of sensitive Coprinus cinereus, by treating
them with a cloned genomic SDC iron-sulfur protein (IP) gene from the resistant
S. commune. The obtained resistance was stably transferred to the progeny.
Key words: flutolanil; drug resistant marker; Coprinus cinereus; Schizophyllum
commune; basidiomycete
-45-
Short Communication
Substantially Complete Removal of Three Major Allergenic Soybean Proteins
(Gly m Bd 30K, Gly m Bd 28K, and the -Subunit of Conglycinin) from Soy
Protein by Using a Mutant Soybean, Tohoku 124
Masahiko Samoto, Yoichi Fukuda, Koji Takahashi,* Kohsei Tabuchi,* Miki Hiemori,** Hideaki Tsuji,** Tadashi Ogawa,** and Yukio Kawamura***,
Central Research Institute, Fuji Oil Co., 4-3 Kinunodai, Yawara-mura,
Ibaraki 300-24, Japan
* Tohoku National Agricultural Experiment Station, Nishisenboku-machi,
Akita 019-21, Japan
** Department of Nutrition, School of Medicine, The University of Tokushima,
Kuramoto-cho 3, Tokushima 770, Japan
*** Protein Science Laboratory, National Food Research Institute, 2-1-2
Kannondai, Tsukuba, Ibaraki 305, Japan
Received June 9, 1997
@A wild-type soybean contains three major allergenic proteins, Gly m Bd
30K, the -subunit of conglycinin, and Gly m Bd 28K. A genetically mutated
soybean (Tohoku 124), which was originally developed as a cultivar lacking
the - and '-subunits of conglycinin, was also found to lack Gly m Bd
28K from immunoblot analysis using monoclonal antibodies specific to Gly
m Bd 28K. This finding indicates the possibility to prepare soy milk and
soy proteins containing none of the three major allergenic soybean proteins
from this cultivar. By applying the previous removal procedure [Samoto
et al., Biosci. Biotech. Biochem., 60, 1911-1913 (1996)] to Tohoku 124,
the substantially complete removal of the three major allergenic proteins
from the soy milk was attained. The removal rates of Gly m Bd 30K, -subunit
of conglycinin, and Gly m Bd 28K were 99.8, 100, and 100%, respectively.
Key words: allergy; soybean protein; soymilk
-46-
Short Communication
A Novel Modification of the Lysine Residue at Position 12 of Histone H4
in Starfish Sperm
Kazuto Nunomura, Takahiko Shimizu, Keiko Hozumi, Toshifumi Takao,* Yasutsugu Shimonishi,* and Susumu Ikegami
Department of Applied Biochemistry, Hiroshima University, 1-4-4 Kagamiyama,
Higashi-hiroshima, Hiroshima 739, Japan
* Institute for Protein Research, Osaka University, Suita, Osaka 565, Japan
Received August 11, 1997
@Post-translational modification of core histones is essential in processes
requiring chromatin remodeling. We report here a novel modification in
histones of the sperm of the starfish, Asterina pectinifera, which involves
an -(-glutamyl)lysine cross-link between the glutamine residue at position
9 of histone H2B and the lysine residue at position 12 of histone H4.
Key words: histone; starfish; transglutaminase
-47-
Short Communication
Simplicissin, a New Pollen Growth Inhibitor Produced by the Fungus, Penicillium
cf. simplicissimum (Oudemans) Thom No. 410
Miyako Kusano, Hiroyuki Koshino,* Jun Uzawa,* Shozo Fujioka,* Tsuyoshi Kawano, and Yasuo Kimura
Department of Agricultural Chemistry, Faculty of Agriculture, Tottori
University, Koyama, Tottori, Tottori 680, Japan
* The Institute of Physical and Chemical Research (RIKEN), Wako-shi, Saitama
351-01, Japan
Received August 29, 1997
@A new pollen growth inhibitor, named simplicissin, was isolated from
Penicillium cf. simplicissimum (Oudemans) Thom No. 410, and its structure
was established by spectroscopic methods including 2D NMR. The biological
activities of the compound were examined by the bioassay methods involving
tea pollen together with lettuce seedlings. The compound inhibited the
growth of the tea pollen tube by 45% at a concentration of 3 mg/liter and
showed complete inhibition at 10 mg/liter.
Key words: Penicillium cf. simplicissimum (Oudemans) Thom No. 410; pollen
growth inhibitor; simplicissin