(Vol.61 No.3 1997)
Structure of Human Chromosome 21-For an
Understanding of Genetic Diseases Including Down's Syndrome-
Kazukiyo Onodera and David Patterson 403
Chemical and Biochemical Characteristics
of Topa Quinone
Minae Mure and Katsuyuki Tanizawa 410
Structural Analyses of Carbon
Chains in 5-Alk(en)ylresorcinols of Rye and Wheat Whole Flour by Tandem
Mass Spectrometry
Yoshikatsu Suzuki, Yasuaki Esumi, Masakazu Uramoto, Yoshiki Kono, and Akira
Sakurai 480
Enantiomeric Resolution of a Germacrene-D
Derivative by Chiral High-performance Liquid Chromatography
Yoshinori Nishii, Taichi Yoshida, and Yoo Tanabe 547
1,5-Dihydroxyiridanyl-lycopene in Tomato
Puree
Tadashi Yokota, Hideo Etoh, Nobuo Ukai, Syunji Oshima, Hideki Sakamoto,and
Yukio Ishiguro 549
Synthesis of 13-Oxo-(Z )-9-octadecenoic Acid and
15-Oxo-(Z )-11-icosenoic Acid, Arrestants of Oryzaephilus surinamensis
L.
Shuhei Nakajima, Aiko Okamura, Taro Takeda, Keiji Sugawara, Masaya
Tateishi, Junkichi Iwasa, and Naomichi Baba 551
Bioactive Gibberellin and Its Metabolism
in Shoots of Phaseolus vulgaris
Tomonobu Toyomasu, Hisakazu Yamane, Noboru Murofushi, Masayuki Katsumi,
and Nobutaka Takahashi 556
Effect of Abscisic Acid on Phosphatidylserine-sensitive
Calcium Dependent Protein Kinase Activity and Protein Phosphorylation in
Rice
Setsuko Komatsu, Hideji Karibe, and Taizo Masuda 418
The Base Specificities of Tomato Ribonuclease
(RNase LE) and Its Asp44 Mutant Enzyme Expressed from Yeast Cells
Kazuko Ohgi, Yasuki Shiratori, Atsushi Nakajima, Masanori Iwama, Hiroko
Kobayashi, Norio Inokuchi, Takashi Koyama, Margret Kock, Andreas Loffler,
Konrad Glund, and Masachika Irie 432
Production of Nigerose, Nigerosyl Glucose,
and Nigerosyl Maltose by Acremonium sp. S4G13
Yutaka Konishi and Kazutoshi Shindo 439
Effects of Degree of Branching on Dispersion
Stability of Phytoglycogen in Aqueous Solution
Koichi Tateishi and Akihiro Nakano 455
Quinoprotein Alcohol Dehydrogenase of Acetic
Acid Bacteria: Kinetic Study on the Enzyme Purified from Acetobacter methanolicus
Jitka Frebortova, Kazunobu Matsushita, Toshiharu Yakushi, Hirohide Toyama,
and Osao Adachi 459
Identification of Essential Ionizable
Groups in Active Site of Aspergillus niger Ώ-Glucosidase
Atsuo Kimura, Akishige Somoto, Haruhide Mori, Osamu Sakai, Hirokazu
Matsui, and Seiya Chiba 475
Biochemical Characterization of
Selective Inhibitors for a Glycyrrhizin-binding Lipoxygenase from Soybeans
Teisuke Furuya, Yoshihito Shimoyama, Nobuyuki Nagata, Tsuyoshi Tomimori,
Toshio Satoh, and Kenzo Ohtsuki 495
Substitutions of Alanine for Cysteine
at a Reactive Thiol Site and for Lysine at a Pyridoxal Phosphate Binding
Site of 1-Aminocyclopropane-1-carboxylate Deaminase
Toyotaka Murakami, Miwa Kiuchi, Hiroyuki Ito, Hirokazu Matsui, and Mamoru
Honma 506
Structural Study of an Exocellular Polysaccharide
of Bacillus circulans
Yuka Isobe, Kumio Yokoigawa, Hiroyasu Kawai, and Yoshiaki Sone 520
Synthesis and Biological Activities
of Ferulic Acid-Amino Acid Derivatives
Zhuang Miao, Hiroshi Kayahara, and Koji Tadasa 527
Inhibitory Effect on the Ώ-Glucosidase
Reaction by the Aggregated State of Sulfoquinovosyldiacylglycerol
Hideyuki Kurihara, Takeshi Mitani, Jun Kawabata, and Mutsuo Hatano
536
Isolation and Identification of
2-O-Methyl-L-rhamnose and 3-O-Methyl-L-rhamnose as Constituents of an Acidic
Polysaccharide of Chlorella vulgaris
Kazutoshi Ogawa, Masanori Yamaura, and Isao Maruyama 539
Gene of LukF-PV-like Component of Panton-Valentine Leukocidin
in Staphylococcus aureus P83 is Linked with lukM
Jun Kaneko, Koji Muramoto, and Yoshiyuki Kamio 541
High-level Expression of Maize C4-type Phosphoenolpyruvate
Carboxylase in Escherichia coli and Its Rapid Purification
Long-Ying Dong, Shingo Hata, and Katsura Izui 545
Inhibitory Activity of Maillard Reaction
Products against Trp-P1-induced Mutagenicity to the Salmonella typhimurium
TA 98 Streptomycin-dependent Strain Assayed in the Absence of S-9 Mix
Akiyoshi Hosono, Usman, and Risako Ohba 424
Mechanism for the Increased Defecation and
Jejunum Mucosal Protein Content in Rats by Feeding Germinated Barley Foodstuff
Osamu Kanauchi, Kazue Agata, and Tohru Fushiki 443
Preventive Effect of Germinated Barley
Foodstuff on Diarrhea Induced by Water-soluble Dietary Fiber in Rats
Osamu Kanauchi, Tomohiko Nakamura, Kazue Agata, and Tohru Fushiki 449
Activity and Activity Coefficient of Water
in Aqueous Solutions and Their Relationships with Solution Structure Parameters
Osato Miyawaki, Akiko Saito, Takeshi Matsuo, and Kozo Nakamura 466
Thermal Properties of Corn Amylose
Incorporating or with Added Free Fatty Acid
Sayuri Akuzawa, Shigeru Sawayama, and Akiko Kawabata 487
Aroma Compounds in the Leaves of Japanese
Pepper (Zanthoxylum piperium DC) and Their Formation from Glycosides
Hiroshi Kojima, Akira Kato, Kikue Kubota, and Akio Kobayashi 491
Comparison between Dietary Soybean Protein
and Casein of the Inhibiting Effect on Atherogenesis in the Thoracic Aorta
of Hypercholesterolemic (ExHC) Rats Treated with Experimental Hypervitamin
D
Masanobu Sakono, Toshihiko Fukuyama, Wei-Hua Ni, Koji Nagao, Hyang-Ran
Ju, Masao Sato, Noriyuki Sakata, Hisao Iwamoto, and Katsumi Imaizumi 514
Changes in Chemical Properties
of Melanoidin by Oxidation and Reduction
Seiichi Homma, Naoko Terasawa, Takako Kubo, Naomi Yoneyama-Ishii, Ko
Aida, and Masao Fujimaki 533
Effect of Vegetable Extracts on Immunoglobulin
Production by Mesenteric Lymph Node Lymphocytes of Sprague-Dawley Rats
Shihoko Kaku, Koji Yamada, Nasra Hassan, Takashi Watanabe, and Michihiro
Sugano 558
Purification and Characterization
of the Family J Catalytic Domain Derived from the Clostridium thermocellum
Endoglucanase CelJ
Mohammad Mainul Ahsan, Miwako Matsumoto, Shuichi Karita, Tetsuya Kimura,
Kazuo Sakka, and Kunio Ohmiya 427
Thiosulfate Reductase from a Moderately
Thermophilic Iron-oxidizing Bacterium, Strain TI-1-Purification and Characterization
Tsuyoshi Sugio, Kimiko Kishimoto, and Keiichi Oda 470
Purification and Properties of Phenylethylamine
Oxidase of Arthrobacter globiformis
Eiichi Shimizu, Keiichi Ohta, Shigeo Takayama, Yuzuru Kitagaki, Katsuyuki
Tanizawa, and Takamitsu Yorifuji 501
Carboxylation Reactions of Pyruvate
: Ferredoxin Oxidoreductase and 2-Oxoglutarate : Ferredoxin Oxidoreductase
from Hydrogenobacter thermophilus TK-6
Ki-Seok Yoon, Masaharu Ishii, Tohru Kodama, and Yasuo Igarashi 510
Microbial Transformation of Carbazole
to Indole-3-acetic Acid by Flavobacterium sp. OCM-1
Hitoshi Obata, Hidehisa Kawahara, and Atsushi Sugiyama 525
Isolation of Extradiol Dioxygenase Genes
That Are Phylogenetically Distant from Other meta-Cleavage Dioxygenase
Genes
Hideto Takami, Toshiaki Kudo, and Koki Horikoshi 530
Clarification of the Promoter Structure of
the Osmoregulated gpd1+ Gene Encoding an Isozyme of NADH-dependent Glycerol-3-phosphate
Dehydrogenase in Fission Yeast
Ryusuke Ohmiya, Hirofumi Aiba, Hisami Yamada, and Takeshi Mizuno 553
Isolation of Propioxatin A from Actinosynnema
sp. SI-23 during a Screening for Serratia piscatorum Metalloproteinase
Inhibitors
Sawao Murao, Satoshi Imafuku, and Hiroshi Oyama 561
A Novel Killer Yeast Effective on Schizosaccharomyces
pombe
Isato Kono and Kunio Himeno 563
-1-
Review
Structure of Human Chromosome 21-For an
Understanding of Genetic Diseases Including Down's Syndrome-
Kazukiyo Onodera and David Patterson*
Department of Agriculture and Life Sciences, Faculty of Agriculture, The
University of Tokyo, Yayoi 1-1, Bunkyo-ku, Tokyo 113, Japan
* Eleanor Roosevelt Institute, 1899 Gaylord Street, Denver, Colorado 80206,
U.S.A.
@We do not intend to give information about the structure of human chromosome
21 in detail but to give a kind of perspective of the present status of
our knowledge about human chromosome 21 in this article. Recent development
of molecular biology has changed revolutionarily the status of our knowledge
of the human chromosome in general. Therefore we will choose a few aspects
of the development in this field and want to discuss how we are able to
use such information to understand the genetic diseases including Down's
syndrome.
Key words: genome structure; human chromosome 21; Down's syndrome
-2-
Review
Chemical and Biochemical Characteristics
of Topa Quinone
Minae Mure and Katsuyuki Tanizawaυ
Department of Biological Science, The Institute of Scientific and Industrial
Research, Osaka University, Ibaraki, Osaka 567, Japan
@2,4,5-Trihydroxyphenylalanine (6-hydroxydopa, abbreviated as topa) is
a perhydroxylated derivative of an amino acid, tyrosine. It rarely occurs
in nature as the free amino acid, but is well known as a very strong neurotoxic
agent. The neurotoxicity is ascribed mainly to its chemical reactivity
and redox capacity. In addition, the oxidized form of topa, topa quinone,
has been shown to be an active non-N-methyl-D-aspartate glutamatergic agonist
and to produce neuronal cell death. Topa and topa quinone may be biosynthesized
enzymatically and non-enzymatically from dopa or from tyrosine through
dopa. On the other hand, topa quinone bound covalently in a protein has
been identified in copper amine oxidase, in which topa quinone serves as
an essential cofactor in catalyzing the oxidation of amines. The topa quinone
cofactor is produced by post-translational modification of a specific tyrosine
precursor with the participation of the enzyme-bound copper ion. These
physiological and biochemical features of topa quinone as well as its chemical
properties revealed by recent model studies are reviewed here.
Key words: copper amine oxidase; 6-hydroxydopa; quinoprotein; topa quinone
-3-
Effect of Abscisic Acid on Phosphatidylserine-sensitive
Calcium Dependent Protein Kinase Activity and Protein Phosphorylation in
Rice
Setsuko Komatsu,υ Hideji Karibe, and Taizo Masuda*
Department of Molecular Biology, National Institute of Agrobiological Resources,
Tsukuba 305, Japan
*Department of Soils and Fertilizers, National Agriculture Research Center,
Tsukuba 305, Japan
Received March 21, 1996
@Protein kinase activity in the embryos of rice (Oryza sativa L. cv. Nipponbare)
during seed soaking in water was assessed under various conditions. The
histone III-S phosphorylating activity in the membrane fraction prepared
from control seedlings treated with water was increased by the addition
of Ca2+ (0.2 mM), and it increased further up on the simultaneous addition
of phosphatidylserine (PS). By contrast, the activity in extracts prepared
from ABA-treated seeds increased by Ca2+ alone and PS did not augment this
increased level of activity. In vitro phosphorylation of the membrane fraction
from control seeds in the presence of Ca2+ and PS resulted in phosphorylation
of two proteins with molecular masses and isoelectric points of 40 kDa/8.9
and 40 kDa/7.6, respectively. When seeds were treated with exogenous ABA,
phosphorylation in vitro of these two proteins was dependent only on the
addition of Ca2+. N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride
(W-7) did not block phosphorylation of these proteins, which is dependent
on Ca2+. The varieties and quantities of phospholipids in the seeds after
soaking in water containing ABA were the same as those after soaking only
in water. Phosphorylation of the 40-kDa proteins in vitro in the membrane
fraction from control seeds required higher Ca2+ concentrations (1-2 mM)
compared to that observed in the membrane fraction from ABA-treated seeds.
Key words: rice embryos; protein phosphorylation; abscisic acid
-4-
Inhibitory Activity of Maillard Reaction
Products against Trp-P1-induced Mutagenicity to the Salmonella typhimurium
TA 98 Streptomycin-dependent Strain Assayed in the Absence of S-9 Mix
Akiyoshi Hosono, Usman, and Risako Ohba
Department of Crop and Animal Science, Faculty of Agriculture, Shinshu
University, Nagano-Minamiminowa 399-45, Japan
Received May 13, 1996
@Of the streptomycin-dependent strains, SD 510 isolated from Salmonella
typhimurium TA 98 provided a sharp response to mutagens without any metabolic
activation; This fact is in contrast to the original test of Ames in which
S-9 mix is usually added for routine screening of mutagens.
@The desmutagenic activity of the Maillard reaction products of glycine
with glucose and of lysine with glucose was studied. The Maillard reaction
products were prepared by heating at various temperatures lower than 120C
for 30 min at pH 7.0, and at 120C for 30 min under various pH conditions,
and their desmutagenic activity against Trp-P1 toward the SD 510 Salmonella
typhimurium TA 98 streptomycin-dependent strain was assayed in the absence
of S-9 mix. This SD strain is effective with its strong response and would
serve as a reliable indicator organism for assessing the desmutagenicity
of Maillard reaction products in the presence or absence of S-9 mix.
Key words: Salmonella typhimurium TA 98 streptomycin-dependent strain;
Trp-P1; Maillard reaction product; desmutagenicity
-5-
Purification and Characterization
of the Family J Catalytic Domain Derived from the Clostridium thermocellum
Endoglucanase CelJ
Mohammad Mainul Ahsan, Miwako Matsumoto, Shuichi Karita,* Tetsuya Kimura,
Kazuo Sakka,υ and Kunio Ohmiya
Department of Bioscience, Faculty of Bioresources and * Center for Molecular
Biology and Genetics, Mie University, Tsu 514, Japan
Received June 21, 1996
@The Clostridium thermocellum endoglucanase CelJ contains two different
catalytic domains in a polypeptide, i.e., a subfamily E1 catalytic domain
and a family J catalytic domain [J. Bacteriol., 178, 5732-5740 (1996)].
The family J catalytic domain (CDJ-CelJ) was produced by a recombinant
Escherichia coli and purified. The purified CDJ-CelJ gave a single band
on SDS-polyacrylamide gel electrophoresis and the molecular weight of this
enzyme (60,000) was consistent with the value (60,333) calculated from
the DNA sequence. CDJ-CelJ hydrolyzed various cellulosic substrates, xylan,
and lichenan but not p-nitrophenyl (PNP)-cellobioside, PNP-glucoside, or
PNP-xyloside at all. CDJ-CelJ was active on Avicel, a microcrystalline
cellulose, and the specific activity of CDJ-CelJ on Avicel (0.0078 U/mg
protein) was comparable to that of CelS, which is recognized as the most
important catalytic subunit of the C. thermocellum, cellulosome, suggesting
that CelJ is also an important catalytic subunit in the cellulosome of
this bacterium, in addition to CelS.
Key words: Clostridium thermocellum; Avicelase; cellulase; endoglucanase
-6-
The Base Specificities of Tomato
Ribonuclease (RNase LE) and Its Asp44 Mutant Enzyme Expressed from Yeast
Cells
Kazuko Ohgi, Yasuki Shiratori,* Atsushi Nakajima,* Masanori Iwama, Hiroko
Kobayashi,* Norio Inokuchi,* Takashi Koyama,* Margret Kock,** Andreas Loffler,**,υυ
Konrad Glund,**,υυυ and Masachika Irie υ
Department of Microbiology, Hoshi College of Pharmacy, Ebara 2-4-41, Shinagawa-ku,
Tokyo 142, Japan
* Department of Microbiology, College of Pharmacy, Nihon University, Narashinodai,
Funabashi 7-7-1, Chiba 271, Japan
** Martin-Luther Universitat, Halle-Wittenberg, Biozentrum der Universitat,
Kurt-Mothes-Straΐe 3, 06120 Halle/Saale, Germany
Received June 24, 1996
@RNase LE from cultured tomato cells is a member of the RNase T2 family.
It is, however, distinguishable from RNase Rh from Rhizopus niveus, a typical
RNase of the RNase T2 family, by its CD spectrum in the 200-250 nm region.
In order to reinvestigate the base specificity of RNase LE and to study
the role of Asn44 in RNase LE, which is considered to correspond to the
base recognition site Asp51 of RNase Rh, RNase LE, and its Asp mutant at
the 44th position were expressed from yeast cells with the same expression
system as RNase Rh [K. Ohgi, et al., J. Biochem., 109, 776-785 (1991)].
@RNase LE with four extra amino acid residues at the 2nd amino acid residue
of mature RNase LE and its Asp44 mutant were secreted from yeast cells
to give a yield of 10 mg/liter and 0.5 mg/liter culture broth, respectively.
The expressed RNase LE (RNase RNAP LE) had the same characteristics as
native RNase LE in the CD spectrum and specific activity. This is the first
example of the expression of plant RNase from microbes and in sufficient
amount to perform further enzymological research.
@The base specificity of RNase LE was guanylic acid preferential and that
of N44D was changed to a more adenylic acid preference as compared to that
of RNase LE. These experiments showed that Asn44 of RNase LE is crucial
for base recognition as the case of Asp51 in RNase Rh, and also suggested
that the base recognition mechanism of RNase LE is very similar to that
of RNase Rh.
Key words: ribonuclease; tomato; expression; base specificity; base recognition
-7-
Production of Nigerose, Nigerosyl
Glucose, and Nigerosyl Maltose by Acremonium sp. S4G13
Yutaka Konishiυ and Kazutoshi Shindo*
Applied Bioresearch Center and * Pharmaceutical Research Laboratories,
Kirin Brewery Co., Ltd., 3 Miyahara-cho, Takasaki-shi, Gunma 370-12, Japan
Received July 11, 1996
@A strain of Acremonium sp. S4G13, which produced an enzyme producing
nigerose, nigerosyl glucose, and nigerosyl maltose from maltooligosaccharides
in high yield, was isolated from soil.
@From these observations, the enzyme was strongly supposed to be a novel
enzyme that hydrolyzed maltooligosaccharides and released glucose, but
also transferred the glucosyl moiety into the C-3 or C-4 position of the
acceptor molecule by Ώ-form in significant amounts.
Key words: Ώ-glucosidase; Acremonium sp. S4G13; nigerose; nigerosyl glucose;
nigerosyl maltose
-8-
Mechanism for the Increased Defecation
and Jejunum Mucosal Protein Content in Rats by Feeding Germinated Barley
Foodstuff
Osamu Kanauchi, Kazue Agata, and Tohru Fushik
Applied Bioresearch Center, Corporate Research and Development Division,
Kirin Brewery Co., Ltd., 3 Miyaharacho, Takasakishi, Gunma 370-12, Japan
* Department of Food Science and Technology, Faculty of Agriculture, Kyoto
University, Oiwakemachi, Kitashirakawa, Sakyo-ku, Kyoto 606, Japan
Received July 15, 1996
@We investigated the effects of germinated barley foodstuff (GBF) on the
fecal excretion and jejunum mucosal protein content in male Sprague-Dawley
rats fed on various diets with the same protein and dietary fiber levels.
Under these experimental conditions, GBF was confirmed to induce greater
fecal output compared with commercial water-soluble or -insoluble dietary
fibers.
@While the dietary fiber extracted from GBF increased the fecal output
and mucosal protein content, the protein fraction of GBF degraded to the
peptide form did not increase the fecal output or mucosal protein content.
Increased mucosal protein and fecal output were thus found to require the
presence of the dietary fiber fraction or possibly the protein fraction
bound tightly to the dietary fiber of GBF. GBF feeding increased the volatile
fatty acids concentration in the cecum, indicating that GBF may be efficiently
fermented in the intestinal tract.
Key words: jejunum mucosa; dietary fiber; germination; volatile fatty acids;
rat
-9-
Preventive Effect of Germinated Barley
Foodstuff on Diarrhea Induced by Water-soluble Dietary Fiber in Rats
Osamu Kanauchi,υ Tomohiko Nakamura, Kazue Agata, and Tohru Fushiki
*
Applied Bioresearch Center, Corporate Research and Development Division,
Kirin Brewery Co., Ltd., 3 Miyaharacho, Takasakishi, Gunma 370-12, Japan
* Department of Food Science and Technology, Faculty of Agriculture, Kyoto
University, Oiwakemachi, Kitashirakawa, Sakyo-ku, Kyoto 606, Japan
Received July 15, 1996
@We investigated the preventive effect of germinated barley foodstuff
(GBF) added to the diet on diarrhea induced by the dietary water-soluble
dietary fibers, polydextrose, hemicellulose, and poly-acrylic acid sodium
salt, in Sprague-Dawley rats. The minimum content of GBF necessary for
blocking diarrhea was 3% (by weight) of the diet.
@Since GBF is mainly derived from the aleurone and scutellum of malted
barley, we assessed the physiological effects of the aleurone and scutellum
fractions derived from barley grains before and after germination. The
addition of fractions containing only germinated barley, and not barley
collected before germination, increased the fecal output and jejunal mucosal
protein content. The effects of malted barley were very similar to those
of GBF.
@It was concluded that germination was necessary to bring about the physiological
effects of GBF. Since non-lignified hemicellulose and Gln-rich protein
were newly synthesized during germination, these might have contributed
to the increased fecal output and jejunal mucosal protein content.
Key words: intestine; dietary fiber; germination; diarrhea; barley
-10-
Effects of Degree of Branching on Dispersion
Stability of Phytoglycogen in Aqueous Solution
Koichi Tateishi and Akihiro Nakano
Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu
University, Minamiminowa, Nagano 399-45, Japan
Received July 19, 1996
@It was supposed that the solubility of (1¨4)(1¨6)-linked Ώ-D-glucans,
e.g., glycogen, phytoglycogen, and amylopectin, in water was related to
the colloidal dispersion stability of such molecules and depended on the
degree of branching. Since phytoglycogen has various degrees of branching
according to the maturation of plant seeds, it was extracted from sweet
corn kernels at several days after pollination (DAP), and we have investigated
the effects of the degree of branching on the dispersion stability of phytoglycogen
(DSP) by salting out, using ammonium sulfate ((NH4)2SO4).
@As the sweet corn kernels matured, the concentration of (NH4)2SO4 needed
for salting out of phytoglycogen increased. According to the degree of
branching measured by periodate oxidation analysis and ΐ-amylolysis, the
fraction of phytoglycogen precipitated at the high concentration with (NH4)2SO4
has a highly branched structure. The turbidity of phytoglycogen aqueous
solution was also measured to discuss the relation between the dispersion
stability of colloidal particles and the degree of branching. We found
that the variation of the degree of branching is closely related to DSP
in an aqueous solution.
Key words: phytoglycogen; salting out; colloidal particle; dispersion stability
-11-
Quinoprotein Alcohol Dehydrogenase of
Acetic Acid Bacteria: Kinetic Study on the Enzyme Purified from Acetobacter
methanolicus
Jitka Frebortova, Kazunobu Matsushita,υ Toshiharu Yakushi, Hirohide
Toyama, and Osao Adachi
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University,
1677-1 Yoshida, Yamaguchi 753, Japan
Received July 24, 1996
@An alcohol dehydrogenase (ADH) complex consisting of subunits I, II,
and III and the free subunit II were purified from Acetobacter methanolicus.
The kinetic parameters of the purified ADH were investigated with several
artificial electron acceptors. Simultaneous reactions with different electron
acceptors showed that these electron acceptors competed with each other.
@Although free subunit II did not show any enzyme activity, part of the
activity was restored after reconstitution with subunit I/III complex purified
from Gluconobacter suboxydans. 2-n-Heptyl-4-hydroxyquinoline-N-oxide (HQNO)
non-competitively inhibited all the reductase activities of native ADH,
while the ferricyanide reductase activity of hybrid ADH was not inhibited
by HQNO but the ubiquinone reductase activity was inhibited competitively.
The kinetic study of native and hybrid ADHs suggests that at least three
heme c moieties are involved in the reduction of ferricyanide and that
the reduction of ubiquinone occurs in subunit II at a site different from
the ferricyanide reacting site.
Key words: quinoprotein; alcohol dehydrogenase; inhibition; reconstitution
-12-
Activity and Activity Coefficient of
Water in Aqueous Solutions and Their Relationships with Solution Structure
Parameters
Osato Miyawaki,υ Akiko Saito, Takeshi Matsuo,υυ and Kozo Nakamura
Department of Applied Biological Chemistry, The University of Tokyo, Bunkyo-ku,
Tokyo 113, Japan
Received August 8, 1996
@The water activity and the activity coefficient of water (ΑW) were calculated
from the freezing point depression by using the Hildebrand and Scott's
equation for various solutions of electrolytes, sugars, alcohols, amide,
and urea. The water activity measured from the freezing point depression
was very close to that at room temperature, showing that the effect of
temperature on the water activity is neglegible. The activity coefficient
of water was described well as a function of the solute mole fraction (XS)
by the following equation. ΑW=exp (ΏX 2(/)S+ΐX 3(/)S)
@The parameter Ώ in this equation correlated well with the B-coefficient
of viscosity for single-valent electrolytes, with the hydration parameter
for nonelectrolytes, and with the number of equatorial-OH groups for sugars
suggesting that the solution structure was reflected in this parameter.
Key words: water activity; activity coefficient of water; solution structure;
hydration; viscosity
-13-
Thiosulfate Reductase from a
Moderately Thermophilic Iron-oxidizing Bacterium, Strain TI-1-Purification
and Characterization
Tsuyoshi Sugio,υ Kimiko Kishimoto, and Keiichi Oda
Department of Biological Function and Genetic Resources Science, Faculty
of Agriculture, Okayama University, 1-1-1 Tsushima Naka, Okayama 700, Japan
Received August 8, 1996
@When grown on Fe2+-medium (pH 1.8) containing the following five L-amino
acids: aspartic acid, glutamic acid, serine, arginine, and histidine, a
moderately thermophilic iron-oxidizing bacterium, strain TI-1, produced
hydrogen sulfide (H2S) outside of the cells and synthesized a novel thiosulfate
reductase, which catalyzed the reduction of thiosulfate with NAD(P)H as
an electron donor to give H2S. The activity of this enzyme in this bacterium
increased 2-fold when elemental sulfur was added to this medium. Thiosulfate
reductase was in the cytosol of the strain and was purified to an electrophoretically
homogeneous state. The apparent molecular weight of thiosulfate reductase
was 230,000 by gel filtration and 58,000 by SDS-PAGE, indicating that the
enzyme is a homotetramer. The enzyme was most active at pH 6.0 and 60C.
Thiosulfate, but not elemental sulfur, sulfite, or tetrathionate, was specifically
used as an electron accepter of this enzyme. Both NADH and NADPH were used
as electron donors. However, NADH was approximately 10 fold superior as
an electron donor to NADPH. Reduced glutathione and mammalian cytochrome
c were not used as electron donors. The Michaelis constants of this enzyme
for thiosulfate, NADH, and NADPH were 0.29, 0.125, and 5.0 mM.
Key words: thiosulfate reductase; iron-oxidizing bacterium; moderately
thermophile; H2S production
-14-
Identification of Essential Ionizable
Groups in Active Site of Aspergillus niger Ώ-Glucosidase
Atsuo Kimura, Akishige Somoto, Haruhide Mori, Osamu Sakai, Hirokazu
Matsui, and Seiya Chiba
Faculty of Agriculture, Hokkaido University, Sapporo 060, Japan
Received August 19, 1996
@A kinetic study was done to identify the ionizable groups in the active
site of Aspergillus niger Ώ-glucosidase (ANGase). From dependence of V
and Km values on pH, we obtained the ionization constants of essential
ionizable groups 1 and 2 of free enzyme; pKe1=3.2 and pKe2=6.4.
@When the
dielectric constant of the reaction mixture was decreased, the pKe1 and
pKe2 were shifted to higher values. The ionization heats (’H 's) of ionizable
groups 1 and 2 were measured to be -0.4 kcal/mol and 0 kcal/mol, respectively.
The water-soluble carbodiimide (WSC), a specific reagent for carboxyl groups,
inactivated the enzyme activity completely, and maltose as substrate decreased
the inactivation. The WSC did not modify the free Cys. These findings suggest
that the essential ionizable groups of ANGase are two kinds of carboxyl
groups: one is a charged type (-COO-, ionizable group 1), and the other
is a protonated type (-COOH, ionizable group 2).
Key words: essential ionizable groups; Ώ-glucosidase; Aspergillus niger;
chemical modification
-15-
Structural Analyses of Carbon
Chains in 5-Alk(en)ylresorcinols of Rye and Wheat Whole Flour by Tandem
Mass Spectrometry
Yoshikatsu Suzuki,υ Yasuaki Esumi, Masakazu Uramoto,* Yoshiki Kono,
and Akira Sakurai
The Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1,
Wako-shi, Saitama 351-01, Japan
* Tamagawa University, Tamagawa Gakuen 6-1-1, Tokyo 194, Japan
Received August 23, 1996
@The locations of double bonds and the linearity of each carbon chain
in 5-alk(en)ylresorcinols present in rye and wheat whole flour were determined
from collision-activated dissociation (CAD) spectra by tandem mass spectrometry.
Among the determined alk(en)ylresorcinols, eleven alkenylresorcinols (C(/)17
to C(/)23) and four alkadienylresorcinols (C(/)19 to C(/)25) had not been
previously identified. In this study, we found that identification of a
radical-anion fragment peak due to a simple allylic cleavage on the methyl
side was essential for determining the locations of double bonds in positionally
isomeric alkenylresorcinols. In addition, an analysis of the CAD spectra
of lithium-adduct cations as precursor ions was useful for determining
the locations of the homoconjugated (Z,Z )-diene group in alkadienylresorcinols.
Key words: 5-alk(en)ylresorcinol; double-bond location; collision-activated
dissociation; charge-remote fragmentation; tandem mass spectrometry
-16-
Thermal Properties of Corn Amylose
Incorporating or with Added Free Fatty Acid
Sayuri Akuzawa, Shigeru Sawayama, and Akiko Kawabata
Department of Nutrition, Faculty of Agriculture, Tokyo University of Agriculture,
1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156,Japan
Received August 26, 1996
@We prepared amylose samples each incorporating one of several free fatty
acids and amylose with a 1% concentration of each added free fatty acid.
We then examined the thermal properties of each sample to compare the results
previously obtained for the thermal properties of starch incorporating
free fatty acids.7) The incorporated free fatty acid value increased with
increasing molecular weight of the amylose component. But the free fatty
acid values were not linearly related to the molecular weight of the amylose
component. The DSC reheating curve for each sample had characteristics
resulting from a different form of recrystallization by each free fatty
acid. It seems that a saturated free fatty acid, which is shorter than
an unsaturated one, could efficiently re-form an amylose/free fatty acid
complex. In contrast, linoleic acid does not form a complex, but coexists
within the amylose chain and/or chain segments.
Key words: corn amylose; amylose/free fatty acid complex; thermal property;
DSC
-17-
Aroma Compounds in the Leaves of Japanese
Pepper (Zanthoxylum piperium DC) and Their Formation from Glycosides
Hiroshi Kojima, Akira Kato, Kikue Kubota,* and Akio Kobayashi*
Food R & D Center, Japan Tobacco Inc., 6-2 Umegaoka, Aoba-ku, Yokohama,
Kanagawa 227, Japan
* Department of Nutrition and Food Science, Ochanomizu University, 2-1-1
Ohtsuka, Bunkyo-ku, Tokyo 112, Japan
Received August 29, 1996
@The volatile compounds in the leaves of Japanese pepper (Zanthoxylum
piperium DC, sanshoh in Japanese) were obtained by steam distillation,
and the potent odorants were evaluated by an aroma extract dilution analysis.
(Z )-3-Hexenol and other C-6 compounds, contributing to the grassy aroma
character, and citronellal and citronellol were most characteristic of
the sanshoh aroma. The glycoside-containing fraction of the methanol extract
was obtained by Amberlite XAD-2 adsorption, and then hydrolyzed by ΐ-glucosidase,
Rohapect D5L, and acetone powder from fresh sanshoh leaves. (Z )-3-Hexenol,
citronellol, benzyl alcohol and 2-phenylethanol were liberated by each
enzyme, while geraniol was liberated by Rohapect D5L. These results suggest
that the characteristic aroma precursors of sanshoh leaves existed as glycosides.
Key words: Japanese pepper; crushed leaf odor; potent odorant; aroma precursor
-18-
Biochemical Characterization
of Selective Inhibitors for a Glycyrrhizin-binding Lipoxygenase from Soybeans
Teisuke Furuya, Yoshihito Shimoyama, Nobuyuki Nagata,* Tsuyoshi Tomimori,**
Toshio Satoh,*** and Kenzo Ohtsuki υ
Laboratory of Genetical Biochemistry, Kitasato University School of Allied
Health Sciences, Sagamihara 228, Japan
* Research Laboratories of Minophagen Pharmaceutical Co., Ltd., Zama, Kanagawa
228, Japan
** School of Pharmacy, Hokuriku University, Ho-3, Kanagawa-machi, Kanazawa
920-11, Japan
*** Faculty of Pharmaceutical Science, Tokushima Bunri University, Yamashiro-cho,
Tokushima 770, Japan
Received September 5, 1996
@By GL-affinity column chromatography (HPLC), a GL-binding lipoxygenase
(gbLOX) was selectively purified from a partially purified soybean LOX-1
fraction. gbLOX was identified as LOX-3 since the partial N-terminal amino
acid sequences of at least four fragments generated from the purified gbLOX
(p96 and p94) digested with trypsin were identical to the corresponding
sequences of soybean LOX-3. The inhibitory effects of various flavonoids
and their modified derivatives, epigallocatechin gallate (EGCG) and a derivative
(oGA) of glycyrrhetinic acid on the activities of two LOXs [gbLOX and ngLOX
(non-GL-binding LOX)] were examined in vitro. It was found that (i) four
compounds [quercetin, EGCG, a derivative (2',4'-DDC) of 3,4-dihydroxychalcone,
and oGA] inhibit gbLOX activity in a dose-dependent manner, but do not
affect ngLOX activity; and (ii) quercetin as well as EGCG effectively prevents
phosphorylation of gbLOX by casein kinase II (CK-II) in vitro. The results
provided here suggest that the potent gbLOX inhibitors function as suppressors
for the LOX-3-catalyzed metabolic pathways in plant cells.
Key words: casein kinase II; 3,4-dihydroxychalcone; epigallocatechin gallate;
glycyrrhizin-binding lipoxygenase; quercetin
-19-
Purification and Properties of
Phenylethylamine Oxidase of Arthrobacter globiformis
Eiichi Shimizu, Keiichi Ohta, Shigeo Takayama, Yuzuru Kitagaki, Katsuyuki
Tanizawa,* and Takamitsu Yorifuji
Department of Bioscience and Biotechnology, Shinshu University, Minamiminowa,
Nagano 399-45, Japan
* The Institute of Scientific and Industrial Research, Osaka University,
Ibaraki, Osaka 567, Japan
Received September 6, 1996
@Phenylethylamine oxidase (EC class 1.4.3) of Arthrobacter globiformis
IFO 12137 (ATCC 8010) was purified to homogeneity. The enzyme had a Mr
of 141,000 and was composed of two apparently identical subunits, which
had a Mr of 71,000 and contained one copper ion. The absorption spectrum
of the enzyme had maxima at 280 and 480 nm, and the ratio A(/)280/A(/)480
was 61.5. Hydrogen peroxide was formed in the oxidation of amines. The
enzyme was most active and stable at pH 6.5. 2-Phenylethylamine and tyramine
were the most active substrates. Several aromatic monoamines, aliphatic
monoamines with 4-11 carbons, higher aliphatic diamines, and histamine
were poor substrates. Benzylamine, putrescine, spermine, and spermidine
were not oxidized. The Kms for 2-phenylethylamine and tyramine were 18
and 85 ΚM, and the V(/)maxs for them were 27.1 and 26.4 Κmol/min/mg of
enzyme, respectively. Benzyl alcohol was a noncompetitive and benzylamine
was a mix-type inhibitor of the enzyme. Carbonyl-blocking reagents such
as methylhydrazine, Cu2+-chelators, HgCl2, p-chloromercuribenzoate, and
Cl- also inhibited the enzyme.
Key words: phenylethylamine oxidase; Arthrobacter globiformis ; coryneform
bacteria; copper-containing amine oxidase; topa quinone
-20-
Substitutions of Alanine for Cysteine
at a Reactive Thiol Site and for Lysine at a Pyridoxal Phosphate Binding
Site of 1-Aminocyclopropane-1-carboxylate Deaminase
Toyotaka Murakami, Miwa Kiuchi, Hiroyuki Ito, Hirokazu Matsui, and Mamoru
Honma
Faculty of Agriculture, Hokkaido University, Sapporo 060, Japan
Received September 10, 1996
@1-Aminocyclopropane-1-carboxylate (ACC) deaminase catalyzes the cyclopropane
ring fragmentation and deamination of ACC. Replacement of cysteine with
alanine at a reactive thiol site, Cys-162, of ACC deaminase did not affect
the enzyme activity, in spite of the previous result that modification
of Cys-162 caused complete loss of the enzyme activity. Substitution of
glycine or valine for the cysteine residue gave a higher Km for ACC without
a significant change of the k0, indicating that changes of the amino acid
side chain had structural effects on substrate binding.
@Replacement of lysine with alanine at the pyridoxal phosphate (PLP) binding
site of the ACC deaminase caused a lower content of PLP and loss of detectable
activity of ACC deamination. This mutant enzyme, K51A, showed absorption
peaks at 330 nm and 405 nm. The peak at 405 nm was shifted to about 425
nm by the addition of ACC, D-, L-alanine, and D-, L-serine. The formation
of aldimine complexes indicated by the spectral shift was reversible. It
is suggested that lysine 51 affects the formation of holoenzyme and is
important in catalysis.
Key words: 1-aminocyclopropane-1-carboxylate deaminase; site directed mutagenesis;
reactive thiol group; pyridoxal phosphate enzyme
-21-
Carboxylation Reactions of Pyruvate
: Ferredoxin Oxidoreductase and 2-Oxoglutarate : Ferredoxin Oxidoreductase
from Hydrogenobacter thermophilus TK-6
Ki-Seok Yoon, Masaharu Ishii, Tohru Kodama,* and Yasuo Igarashi υ
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku,
Tokyo 113, Japan
* Department of Textile Science and Technology, Shinshu University, Tokida
3-15-1, Ueda-shi, Nagano 386, Japan
Received September 11, 1996
@Enzymatic reactions involving pyruvate : ferredoxin oxidoreductase and
2-oxoglutarate : ferredoxin oxidoreductase from a thermophilic, aerobic,
chemolithoautotrophic, and hydrogen-oxidizing bacterium, Hydrogenobacter
thermophilus TK-6, were investigated as the CO2 exchange reaction and CO2
fixation reaction using ferredoxin isolated from the same organism as a
reductant.
@The reduced ferredoxin was required in the pyruvate synthetic reaction
and the 2-oxoglutarate synthetic reaction by a cell extract that had been
treated with a PD-10 column to remove the low molecular weight substances.
[14C]Pyruvate and [14C]2-oxoglutarate were detected as products of pyruvate
synthetic and 2-oxoglutarate synthetic reactions, respectively. Further
evidence for the operation of pyruvate : ferredoxin oxidoreductase and
2-oxoglutarate : ferredoxin oxidoreductase was obtained from experiments
on CO2 exchange reactions using the purified enzymes.
Key words: reductive TCA cycle; hydrogen bacterium; pyruvate : ferredoxin
oxidoreductase; 2-oxoglutarate : ferredoxin oxidoreductase
-22-
Comparison between Dietary Soybean Protein
and Casein of the Inhibiting Effect on Atherogenesis in the Thoracic Aorta
of Hypercholesterolemic (ExHC) Rats Treated with Experimental Hypervitamin
D
Masanobu Sakono, Toshihiko Fukuyama, Wei-Hua Ni, Koji Nagao, Hyang-Ran
Ju, Masao Sato, Noriyuki Sakata,* Hisao Iwamoto,** and Katsumi Imaizumiυ
Department of Food Science and Technology, Faculty of Agriculture, Kyushu
University, Fukuoka 812-81, Japan
* Faculty of Medicine, Fukuoka University, Fukuoka 814-80, Japan
** Department of Animal Science, Faculty of Agriculture, Kyushu University,
Fukuoka 812-81, Japan
Received September 24, 1996
@Atherosclerotic lesions of the thoracic aorta were induced in exogenously
hypercholesterolemic (ExHC) rats by treating initially with hypervitamin
D2 and subsequently feeding on hypercholesterolemic diets for 180 days.
Dietary soybean protein, in comparison with casein, substantially decreased
the degree of atherosclerotic lesions, which was evaluated by intimal thickening,
although with a similar topographical distribution. The casein-fed rats
tended to maintain a high concentration of serum cholesterol, particularly
in triacylglycerol-rich lipoproteins. The concentrations of apo A-I and
TBARS in the serum was comparable between the dietary protein groups. The
data suggest that dietary soybean protein, compared to casein, produced
lipoproteins which were less atherosclerotic by partitioning cholesterol
in the triacylglycerol-poor lipoproteins.
Key words: exogenous hypercholesterolemia; cholesterol; atherosclerosis;
soybean protein; casein; hypervitaminosis D2
-23-
Structural Study of an Exocellular Polysaccharide
of Bacillus circulans
Yuka Isobe,υ Kumio Yokoigawa, Hiroyasu Kawai, and Yoshiaki Sone*
Department of Food Science and Nutrition, Nara Women's University, Nara
630, Japan
* Department of Human Life Science, Osaka City University, Osaka 558, Japan
Received October 2, 1996
@Bacillus circulans, a soil bacterium, produced an exocellular polysaccharide
of high viscosity. On the basis of the results of methylation analysis,
mild acid hydrolysis, and 1D and 2D 1H-NMR spectroscopy, it was concluded
that the polysaccharide has a basic repeating unit composed of ΐ-L-rhamnopyranose,
Ώ-D-mannopyranose, Ώ-D-galactopyranose, and Ώ-D-glucopyranuronic acid
with the following structure: ΐ-L-Rha p 1.51 1.5« "
Έ4 1.5¨3-Ώ-D-GlcUA
p1¨2-Ώ-D-Man p1¨3-Ώ-D-Gal p1¨
Key words: Bacillus circulans ; exo-polysaccharide; 1H-NMR
-24-
Note
Microbial Transformation of Carbazole
to Indole-3-acetic Acid by Flavobacterium sp. OCM-1
Hitoshi Obata,υ Hidehisa Kawahara, and Atsushi Sugiyama
Department of Biotechnology, Faculty of Engineering, Kansai University,
Suita-shi, Osaka 564, Japan
Received June 28, 1996
@A carbazole-using bacterium was isolated from oil polluted soil and identified
as Flavobacterium sp. OCM-1 from its taxonomical characteristics. Its optimal
culture conditions were identified. The growth and carbazole-degradation
were found in the ranges of 20-30C and pH 6-8. We found microbial production
of indole-3-acetic acid from carbazole by strain OCM-1. Indole-3-acetic
acid was identified as a metabolite of carbazole using thin-layer chromatography,
mass and 1H-NMR-spectra. 1.5 mg of indole-3-acetic acid was formed from
250 mg of carbazole.
Key words: Flavobacterium sp. OCM-1; carbazole; biodegradation; indole-3-acetic
acid
-25-
Note
Synthesis and Biological Activities
of Ferulic Acid-Amino Acid Derivatives
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 July 17, 1996
@Three series of ferulic acid derivatives (feruloylaminoacid benzyl ester
or methyl ester, feruloylaminoacid, and 4-O-[N-(carbobenzyloxy)-aminoacyl]ferulic
acid) were synthesized newly, and their superoxide dismutase (SOD)-like,
platelet aggregation (PA)-inhibitory, tyrosinase-inhibitory, and angiotensin
converting enzyme (ACE)-inhibitory activities were evaluated. 4-O-[N-(Carbobenzyloxy)isoleucyl]ferulic
acid (20) and 4-O-[N-(carbobenzyloxy)prolyl]ferulic acid (21) showed potent
PA-inhibitory activities and tyrosinase-inhibitory activities, while they
also had SOD-like activities at the same level as ferulic acid.
Key words: ferulic acid-amino acid derivatives; superoxide radical; platelet
aggregation; tyrosinase; angiotensin converting enzyme
-26-
Note
Isolation of Extradiol Dioxygenase
Genes That Are Phylogenetically Distant from Other meta-Cleavage Dioxygenase
Genes
Hideto Takami,* Toshiaki Kudo,υ and Koki Horikoshi*
JRDC/MSU-Microbial Evolution Project, Microbiology Laboratory, RIKEN Institute,
2-1 Hirosawa, Wako, Saitama 351-01, Japan
Received August 8, 1996
@meta-Cleavage dioxygenase genes were isolated from the benzoate-assimilating
bacteria Pseudomonas stutzeri A401 and Pseudomonas mendocina KC37. The
gene products had the enzymatic activity of 2,3-dihydroxy-biphenyl-1,2-dioxygenase.
Phylogenetic analysis based on the alignment of amino acid sequences suggests
that these enzymes are distantly related to other meta-cleavage enzymes
and may be members of a new family of extradiol dioxygenases.
Key words: Pseudomonas stutzeri; Pseudomonas mendocina; extradiol dioxygenase
-27-
Note
Changes in Chemical Properties
of Melanoidin by Oxidation and Reduction
Seiichi Homma, Naoko Terasawa,* Takako Kubo, Naomi Yoneyama-Ishii, Ko
Aida, and Masao Fujimaki
Department of Nutrition and Food Science, Ochanomizu University, 2-1-1
Ohtsuka, Bunkyo-ku, Tokyo 112, Japan
* Faculty of Education, Kanazawa University, Kakuma-machi, Kanazawa-shi,
Ishikawa 920-11, Japan
Received August 8, 1996
@Nondialyzable model melanoidin prepared from glucose and glycine was
oxidized with potassium ferricyanide or reduced with sodium borohydride.
Compared with intact melanoidin, the oxidation reduced the pI value and
increased the chelating activity, while reduction had the completely opposite
effect. Both reactions on melanoidin increased the molecular weight and
reduced the E 1%(/)500 nm value.
Key words: melanoidin; oxidation of melanoidin; reduction of melanoidin;
electrofocusing of melanoidin; reductone
-28-
Note
Inhibitory Effect on the Ώ-Glucosidase
Reaction by the Aggregated State of Sulfoquinovosyldiacylglycerol
Hideyuki Kurihara,* Takeshi Mitani,* Jun Kawabata,** and Mutsuo Hatano*
* Faculty of Fisheries, Hokkaido University, Minato, Hakodate, Hokkaido
041, Japan
** Faculty of Agriculture, Hokkaido University, Kita-ku, Sapporo, Hokkaido
060, Japan
Received August 8, 1996
@The inhibitory activity of aggregated SQDG toward the Ώ-glucosidase
reaction decreased with increasing turbidity of the SQDG suspension. Dideacylated
derivatives of SQDG, sulfoquinovosylglycerol and sulfoquinovose, showed
no inhibition toward the reaction. The expression of inhibition by the
SQDG family would require a hydrophobic acyl group(s) to produce an aggregate
and/or interact with a hydrophobic site in the enzyme.
Key words: SQDG; inhibition; Ώ-glucosidase; aggregation
-29-
Note
Isolation and Identification
of 2-O-Methyl-L-rhamnose and 3-O-Methyl-L-rhamnose as Constituents of an
Acidic Polysaccharide of Chlorella vulgaris
Kazutoshi Ogawa, Masanori Yamaura, and Isao Maruyama*
Department of Fundamental Science, College of Science and Engineering,
Iwaki Meisei University, Iwaki, Fukushima 970, Japan
* Chlorella Industry Co., Ltd., Chikugo, Fukuoka 833, Japan
Received August 14, 1996
@The monosaccharides 2-O-methyl-L-rhamnose and 3-O-methyl-L-rhamnose were
isolated and identified as components of the acidic polysaccharide of Chlorella
vulgaris K-22. 2-O-Methyl-L-rhamnose was first found in algae.
Key words: Chlorella vulgaris; acidic polysaccharide; 2-O-methyl-l-rhamnose;
3-O-methyl-l-rhamnose
-30-
Note
Gene of LukF-PV-like Component of Panton-Valentine
Leukocidin in Staphylococcus aureus P83 is Linked with lukM
Jun Kaneko, Koji Muramoto, and Yoshiyuki Kamio υ
Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku
University, 1-1 Tsutsumi-dori Amamiyamachi, Aoba-ku, Sendai 981, Japan
Received August 23, 1996
@Staphylococcus aureus P83 (ATCC 31890) produces five components, I to
V for synergistic toxins, leukocidin and Α-hemolysin [Sudo et al., Biosci.
Biotech. Biochem., 56, 1786-1789 (1995)].
@We report here the identification
of component II, which is designated LukF-PV(P83) and its gene (lukF-PV(P83)).
The lukF-PV(P83) gene was found to be one base downstream of the stop codon
of the lukM gene. The deduced amino acid sequence of LukF-PV(P83) showed
78.4% identity with that of LukF-PV. The lukM and lukF-PV(P83) genes were
encoded as one operon like that of Panton-Valentine leukocidin.
Key words: Staphylococcus aureus P83; leukocidin; LukF-PV; LukS-PV; LukM
-31-
Note
High-level Expression of Maize C4-type Phosphoenolpyruvate
Carboxylase in Escherichia coli and Its Rapid Purification
Long-Ying Dong, Shingo Hata, and Katsura Izuiυ
Laboratory of Applied Botany, Faculty of Agriculture, Kyoto University,
Sakyo-ku, Kyoto, Kyoto 606-01, Japan
Received August 26, 1996
@Maize C4-type phosphoenolpyruvate carboxylase (PEPC) was expressed in
E. coli with the pET32 system. The expressed fusion PEPC was active and
its amount comprised more than 10% of total soluble protein. The specific
activity increased by about 45-fold, compared with our previous system
[S. Yanagisawa and K. Izui, Agric. Biol. Chem., 54, 241-243 (1990)]. The
fusion PEPC was rapidly purified with HisEbind metal chelation resin,
showing a single band on SDS-PAGE.
@Moreover, the tag domain fused at the
N-terminus did not have any effect on catalytic and regulatory properties
of PEPC.
Key words: phosphoenolpyruvate carboxylase; expression; purification; pET32
plasmid
-32-
Note
Enantiomeric Resolution of a Germacrene-D
Derivative by Chiral High-performance Liquid Chromatography
Yoshinori Nishii, Taichi Yoshida, and Yoo Tanabeυ
School of Science, Kwansei Gakuin University, 1-1-155 Uegahara, Nishinomiya,
Hyogo 662, Japan
Received August 28, 1996
@Direct enantiomeric resolution of the germacrene-D derivative, (5E,7S
*,9S *)-9-hydroxy-7-isopropyl-4,10-bis(methylene)-5-cyclo decen-1-one,
was carried out by HPLC with a chiral stationary phase. This method showed
that germacrene-D contained in ylang ylang oil was 98% e.e., whereas germacrene-D
extracted from Solidago altissima L. was almost racemic.
Key words: germacrene-D; resolution; optical purity; chiral HPLC
-33-
Note
1,5-Dihydroxyiridanyl-lycopene in Tomato
Puree
Tadashi Yokota, Hideo Etoh,υ Nobuo Ukai,* Syunji Oshima,* Hideki Sakamoto,*
and Yukio Ishiguro*
Department of Applied Biological Chemistry, Shizuoka University, 836 Ohya,
Shizuoka 422, Japan
* Kagome Co., Ltd., 17 Nishitomiyama, Nishinasuno-cho, Nasu-gun, Tochigi
329-27, Japan
Received August 29, 1996
@1,5-Dihydroxyiridanyl-lycopene was isolated from tomato puree. The oxidation
of lycopene by hydrogen peroxide also afforded 1,5-dihydroxyiridanyl-lycopene,
in addition to lycopene-1,2-epoxide and 1,5-epoxyiridanyl-lycopene. A reaction
pathway for this dihydroxy compound is proposed.
Key words: 1,5-dihydroxyiridanyl-lycopene; tomato puree; oxidation of lycopene;
lycopene-5,6-epoxide; 5,6-dihydroxy-5,6-dihydrolycopene
-34-
Note
Synthesis of 13-Oxo-(Z )-9-octadecenoic Acid
and 15-Oxo-(Z )-11-icosenoic Acid, Arrestants of Oryzaephilus surinamensis
L.
Shuhei Nakajima, Aiko Okamura, Taro Takeda, Keiji Sugawara, Masaya Tateishi,
Junkichi Iwasa, and Naomichi Baba
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University,
1-1-1 Tsushimanaka, Okayama 700, Japan
Received August 30, 1996
@Two arrestants of the sawtoothed grain beetle (Oryzaephilus surinamensis
L.), 13-oxo-(Z )-9-octadecenoic acid and 15-oxo-(Z )-11-icosenoic acid,
were synthesized for the first time by utilizing a Z-selective Wittig reaction.
Key words: Oryzaephilus surinamensis L.; arrestant; 13-oxo-(Z)-9-octadecenoic
acid; 15-oxo-(Z)-11-icosenoic acid
-35-
Note
Clarification of the Promoter Structure
of the Osmoregulated gpd1+ Gene Encoding an Isozyme of NADH-dependent Glycerol-3-phosphate
Dehydrogenase in Fission Yeast
Ryusuke Ohmiya, Hirofumi Aiba, Hisami Yamada, and Takeshi Mizuno υ
Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University,
Chikusa-ku, Nagoya 464, Japan
Received September 11, 1996
@The gpd1+ gene of Schizosaccharomyces pombe, encoding an isozyme of NADH-dependent
glycerol-3-phosphate dehydrogenase, is responsible for an osmoinducible
glycerol production in response to external high osmotic stimuli, and its
expression is osmoregulated at the level of transcription. In this study,
the structure of the osmoinducible promoter of gpd1+ is described. Not
only the core promoter elements including the putative TATA-box and transcription
start site, but also a short upstream activation sequence (UAS) was identified,
which are involved in the osmotic induction of the gpd1+ gene through the
Wis1 MAP (mitogen activated protein) kinase pathway.
Key words: glycerol synthesis; osmoregulation; MAP-signaling pathway; upstream
activation sequence; Schizosaccharomyces pombe
-36-
Note
Bioactive Gibberellin and Its Metabolism
in Shoots of Phaseolus vulgaris
Tomonobu Toyomasu,υ Hisakazu Yamane,* Noboru Murofushi, Masayuki Katsumi,**
and Nobutaka Takahashi
Department of Applied Biological Chemistry, The University of Tokyo, 1-1-1
Yayoi, Bunkyo-ku, Tokyo 113, Japan
* Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi,
Bunkyo-ku, Tokyo 113, Japan
** Biology Department, International Christian University, Osawa, Mitaka,
Tokyo 181, Japan
Received September 13, 1996
@Evidence is presented for gibberellin A1 (GA1) being bioactive per se
in regulating the shoot elongation of Phaseolus vulgaris. It is also suggested
that the lower response of cv. Masterpiece (dwarf ) to GA1 than of cv.
Kentucky Wonder (normal) was not due to any difference in GA1 metabolism,
but to a defect in the sequence of events from the reception of GA1 to
elongation.
Key words: GA1 metabolism; prohexadione; shoot elongation; Phaseolus vulgaris
L.; dwarfism
-37-
Note
Effect of Vegetable Extracts on Immunoglobulin
Production by Mesenteric Lymph Node Lymphocytes of Sprague-Dawley Rats
Shihoko Kaku, Koji Yamada, Nasra Hassan, Takashi Watanabe, and Michihiro
Sugano
Laboratory of Food Science, Department of Food Science and Technology,
Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku,
Fukuoka 812-81, Japan
Received September 26, 1996
@To clarify the immunoglobulin production-regulating activity of vegetable
extracts, mesenteric lymph node lymphocytes of Sprague-Dawley rats were
cultured in the presence of 25 different vegetable extracts. The immunoglobulin
content in the culture medium determined by ELISA indicated that the lily
family (Liliaceae) vegetables most strongly enhanced the production of
IgA and IgG, whereas they suppressed IgE production.
Key words: food allergy; IgA antibody; IgE antibody; lily family; vegetable
-38-
Note
Isolation of Propioxatin A from
Actinosynnema sp. SI-23 during a Screening for Serratia piscatorum Metalloproteinase
Inhibitors
Sawao Murao,υ Satoshi Imafuku, and Hiroshi Oyama
Department of Applied Microbial Technology, The Kumamoto Institute of Technology,
Ikeda 4-22-1, Kumamoto 860, Japan
Received October 3, 1996
@An inhibitor of Serratia metalloproteinase was found in the cultured
broth of strain SI-23, which was identified to be Actinosynnema sp. The
inhibitor had a potent inhibitory effect on Serratia metalloproteinase
and a weak effect on thermolysin, but no effect on other groups of proteinases.
The structure of the inhibitor was deduced to be propioxatin A, reported
as an enkephalinase B inhibitor.
Key words: metalloproteinase; proteinase inhibitor; Serratia piscatorum;
Actinosynnema; propioxatin A
-39-
Note
A Novel Killer Yeast Effective on Schizosaccharomyces
pombe
Isato Kono and Kunio Himeno
Industrial Technology Center of Okayama Prefecture, 5301 Haga, Okayama-shi,
Okayama 701-12, Japan
Received October 7, 1996
@To control the extent of deacidification in wine making, we screened
Kluyveromyces strains by their activity to kill the fission yeast Schizosaccharomyces
pombe. Among Kluyveromyces IFO strains tested, K. waltii IFO 1666T was
shown to have the desired activity. The killer spectrum of this strain
was different from those of the other known killer yeasts. The activity
was found in the culture medium and was lost by protease treatment. The
activity was associated with the precipitate obtained by an increase of
ammonium sulfate concentration. The toxin was larger than 10,000 daltons
as judged by ultrafiltration.
Key words: killer; yeast; Schizosaccharomyces pombe; Kluyveromyces