(Vol.61 No.11 1997)
Effects of Lecithin Addition in Oil or Water Phase on the
Stability of Emulsions Made with Whey Proteins
Yukiko Yamamoto and Megumi Araki
Genetic Analysis of Plasmid-specific Pheromone Signaling
Encoded by pPD1 in Enterococcus faecalis
Jiro Nakayama and Akinori Suzuki
Contents of Resveratrol, Piceid, and Their Isomers in Commercially
Available Wines Made from Grapes Cultivated in Japan
Michikatsu Sato, Yumiko Suzuki, Tohru Okuda, and Koki Yokotsuka 1800
Specific Interaction of Cytokinins and Their Analogs with
Rotenone-sensitive Internal NADH Dehydrogenase in Potato Tuber Mitochondria
Masayuki Sue, Hideto Miyoshi, and Hajime Iwamura 1806
Different Responses of Apolipoprotein A-I, A-IV, and B
Gene Expression during
Kei Sonoyama and Yoritaka Aoyama 1810
Metabolic Behavior of Angiotensins in Normotensive Human
Plasma in the Supine and Upright Postures
Toshiro Matsui, Hiroshi Matsufuji, Kei Tamaya, Terukazu Kawasaki, Yutaka
Osajima
Involvements of Trp23 in the Chitin-binding and of Trp131
in the Chitinase Activity of Rye Seed Chitinase-a
Takeshi Yamagami and Gunki Funatsu
Water Permeability of Plasma Membranes of Cultured Rice,
Grape, and CH27 Cells Measured Dielectrically
Eisuke Ishikawa, Osato Miyawaki, Kozo Nakamura
Dielectric Relaxation of Aqueous Solution with Low-molecular-weight
Nonelectrolytes and Its Relationship with Solution Structure
Akiko Saito, Osato Miyawaki, and Kozo Nakamura 1831
A Model System for Convenient Fluorescent Labeling of
Sugar Chain in Taka-amylase A
Mikihiko Kobayashi, Yutaka Sasaki, and Yogo Chiba
Isolation of Taka-amylase A Peptides Required for Substrate
Binding
Yutaka Sasaki, Hidenari Takahara, and Mikihiko Kobayashi 1840
Growth Factors for a Primary Chick Muscle Cell Culture
from Shochu Distillery
Luthfi D. Mahfudz, Kazuki Nakashima, Akira Ohtsuka, Kunioki Hayashi
Nigrosporins A and B, New Phytotoxic and Antibacterial
Metabolites Produced by a Fungus Nigrospora oryzae
Masayasu Tanaka, Toshiro Fukushima, Yasuko Tsujino, and Takane Fujimori
1848
Cloning and Sequence Analysis of the Gene (alyII ) Coding
for an Alginate Lyase of Pseudomonas sp. OS-ALG-9
Jintana Kraiwattanapong, Toshihiko Ooi, and Shinichi Kinoshita
Secretion of Human Interleukin-2 in Biologically Active
Form by Bacillus brevis Directly into Cultute Medium
Yasushi Takimura, Masashi Kato, Toshihiro Ohta, Hideo Yamagata, and Shigezo
Udaka 1858
Degree of Polymerization of Cellulose from Acetobacter
xylinum BPR2001 Decreased by Cellulase Produced by the Strain
Naoki Tahara, Mari Tabuchi, Kunihiko Watanabe, Hisato Yano, Yasushi Morinaga,
Fumihiro Yoshinaga
Structural Analysis of N-Glycans of Storage Glycoproteins
in Soybean (Glycine max. L) Seed
Yoshinobu Kimura, Akira Ohno, Shigeaki Takagi
Synthesis, Biological Activity, and Metabolism of 8',8',8'-Trideuteroabscisic
Acid
Yasushi Todoroki, Sei-ichi Nakano, Nobuhiro Hirai, Toshiaki Mitsui, and
Hajime Ohigashi 1872
Hyperproduction of L-Threonine by an Escherichia coli
Mutant with Impaired L- Threonine Uptake
Kazuyuki Okamoto, Kuniki Kino, and Masato Ikeda 1877
Activation of Macrophages by Sulfated Glycopeptides in
Ovomucin, Yolk Membrane, and Chalazae in Chicken Eggs
Hiroko Tanizaki, Hiroki Tanaka, Hiroyuki Iwata, and Akio Kato 1883
Purification and Characterization of Cystine Lyase a from
Broccoli Inflorescence
Koji Ukai and Jiro Sekiya 1890
Effects of Plant Growth Regulators on Shoot Growth and
Flowering of a Perennial Paddy Weed, Sagittaria pygmaea Miq.
Takumi Yoshimura, Hitoshi Kuramochi, Makoto Konnai, Hideharu Seto, Takeshi
Sassa, and Koichi Yoneyama 1896
Protective Effect of Green Tea Extract and Tea Polyphenols
against the Cytotoxicity of 1,4-Naphthoquinone in Isolated Rat Hepatocytes
Chika Miyagawa, Chen Wu, David Opare Kennedy, Teruyo Nakatani, Kimiko Ohtani,
Senji Sakanaka, Mujo Kim, and Isao Matsui-Yuasa 1901
(E )-2-(4'-Methyl-3'-pentenylidene)-4-butanolide, Named
-Acariolide: A New Monoterpene Lactone from the Mold Mite, Tyrophagus
putrescentiae (Acarina: Acaridae)
Atsushi Morino, Yasumasa Kuwahara, Sigeru Matsuyama, and Takahisa Suzuki
1906
Immunopotentiating Activity of the Water-soluble Lignin
Rich Fraction Prepared from LEM-The Extract of the Solid Culture Medium
of Lentinus edodes Mycelia-
Yoshiki Yamamoto, Hiroyuki Shirono, Keiko Kono, and Yasuhiro Ohashi
Enzymatic Properties of Double Mutant Enzymes at Asp51
and Trp49 and Asp51 And Tyr57 of RNase Rh from Rhizopus niveus
Kazuko Ohgi, Mitsuaki Takeuchi, Masanori Iwama, and Masachika Irie
Structural Property and in Vitro Self-assembly of Shark
Type I Collagen
Yoshihiro Nomura, Masaya Yamano, Chikayuki Hayakawa, Yasuhiro Ishii,
and Kunio Shirai 1919
Note
Growth Suppressing Activity for Endothelial Cells Induced from Macrophages
by Carboxymethylated Curdlan
Shigeyuki Usui, Toshiyuki Matsunaga, Shigeo Ukai, and Tadashi Kiho 1924
Note
Lipase-catalyzed Synthesis of Arbutin Cinnamate in an Organic Solvent and
Application of Transesterification to Stabilize Plant Pigments
Nobuyoshi Nakajima, Kohji Ishihara, Shingo Matsumura, Hiroki Hamada,
Kaoru Nakamura, and Tsutomu Furuya 1926
Note
New Acylated Anthocyanins from Brassica campestris var. chinensis
Masahiro Suzuki, Tadahiro Nagata, and Norihiko Terahara
Note
Manufacturing High Purity Maltose and Maltotetraose from Starch by a Novel
and
Noriyuki Tachikake, Hirotsugu Ogawa, and Masaya Toda 1931
Note
Heterodimeric Aminopeptidase A from Bacillus licheniformis NS115
Tae-Kwang Oh, Mi-Ja Park, Jung-Kee Lee, Hyung-Kwoun Kim, and Hee-Sop
Nam
Note
Stimulation of Tumor Necrosis Factor and Interleukin-1 Productivity by
the Oral
Wataru Komatsu, Kazumi Yagasaki, Yutaka Miura, and Ryuhei Funabiki 1937
Note
An Established Hybridoma Clone Producing a Monoclonal Antibody against
Vibrio anguillarum
Koji Ikura, Shinsuke Kodama, Hiroyuki Hashimoto, Kaeko Hayashi, Hajime
Mori, Masatoshi Ichida, Minoru Sorimachi, Ken-ichi Kudo, and Saburo Hara
1939
Note
Antioxidant Activity of Ferulic Acid -Glucuronide in the LDL Oxidation
System
Takeo Ohta, Tomoko Nakano, Yukari Egashira, and Hiroo Sanada 1942
Note
Two-mode Analysis by High-performance Liquid Chromatography of -Aminobenzoic
Ethyl Ester-derivatized Monosaccharides
Shoichi Yasuno, Takeomi Murata, Kazuko Kokubo, Takashi Yamaguchi, and Masugu
Kamei
Note
Photocatalysis-dependent Inactivation of Lactobacillus Phage PL-1 by a
Ceramics Preparation
Yukari Kakita, Nobuhiro Kashige, Fumio Miake, and Kenji Watanabe
Note
Human Placental Fructose-6-phosphate,2-kinase/Fructose-2,6-bisphosphatase:
Its Isozymic Form, Expression and Characterization
Ryuzo Sakakibara, Mika Uemura, Takafumi Hirata, Noriko Okamura, and Mie
Kato 1949
Note
Nitric Oxide Formation by Macrophages Stimulated with Water Extracts from
Meats and Offals
Misao Miwa, Kiyohiro Shibata, Kiyomi Nagayama, and Katuhiro Aikawa
Note
Synthesis of Asymmetrically Labeled Sucrose by a Recombinant Sucrose Synthase
Tomonori Nakai, Naoto Tonouchi, Takayasu Tsuchida, Hitoshi Mori, Fukumi
Sakai, and Takahisa Hayashi 1955
Note
Biodegradabilities of Ethylenediamine-N,N '-disuccinic Acid (EDDS) and
Other Chelating Agents
Rikiya Takahashi, Naoshi Fujimoto, Masaharu Suzuki, and Takakazu Endo 1957
Short Communication
Panton-Valentine Leukocidin Genes in a Phage-like Particle Isolated from
Mitomycin C-Treated Staphylococcus aureus V8 (ATCC 49775)
Jun Kaneko, Takahiro Kimura, Yoshiyuki Kawakami, Toshio Tomita, and Yoshiyuki
Kamio 1960
-1-
Effects of Lecithin Addition in Oil or Water Phase on the
Stability of Emulsions Made with Whey Proteins
Yukiko Yamamoto and Megumi Araki
Department of Food and Nutrition, Faculty of Human Life Science, Osaka City University, Sugimoto 3-chome, Sumiyoshi-ku, Osaka 558, Japan
Received January 29, 1997
@The effects of lecithin addition in oil or water phase on the stability
of oil-in-water emulsions made with 0.1 wt% whey protein and 10 wt% n-tetradecane
at neutral and acidic pH were studied by monitoring the gravitational creaming
and phase separation. The effects of lecithin addition on the interfacial
behavior of -lactoglobulin were also studied to compare with the results
of emulsion stability. At neutral pH, crude phosphatidylcholine (PC) from
egg yolk or soybean increased the stability of the emulsion made with protein
and lowered the interfacial tension of protein films more effectively than
pure egg PC. A more remarkable effect on both the emulsion stability and
the interfacial tension was found when crude PC was added in the oil phase
rather than in the water phase. The purity of lecithins and the way to
add them are suggested to be very important to make a stable emulsion with
protein. On acidic pH (4.5 or 3.0), the increased creaming or phase separation
in a whey protein-stabilized emulsion, but the lowered interfacial tension
of -lactoglobulin films, were found upon the addition of pure or crude
PC in oil or water phase. These results suggest that in acidic pH, densely
packed films may be formed on a planar oil-water interface, but not on
adsorbed layers around oil droplets in an emulsion.
Key words: emulsion stability; interfacial tension; whey protein; -lactoglobulin;
lecithin
-2-
Genetic Analysis of Plasmid-specific Pheromone Signaling
Encoded by pPD1 in Enterococcus faecalis
Jiro Nakayama and Akinori Suzuki
Department of Applied Biological Chemistry, The University of Tokyo, Bunkyo-ku, Tokyo 113, Japan
Received March 14, 1997
@Certain plasmids in Enterococcus faecalis encode a mating response to
recipient-produced peptide sex pheromones. Targeted disruption of tra genes
on pPD1 suggested that TraA plays a central role in the plasmid-specific
pheromone signaling pathway. TraA functioned as a negative regulator for
the pheromone-inducible conjugal transfer. Complementation analysis of
pPD1 tra gene mutants by pAD1 suggested that the pheromone binding function
of TraC was non-specific between these plasmids, but the function of TraA
and the pheromone shutdown function of TraB are plasmid-specific.
Key words: Enterococcus faecalis; bacterial sex pheromone; tra genes; pheromone
signaling; targeted gene disruption
-3-
Contents of Resveratrol, Piceid, and Their Isomers in Commercially
Available Wines Made from Grapes Cultivated in Japan
Michikatsu Sato, Yumiko Suzuki, Tohru Okuda,* and Koki Yokotsuka*
Wines & Spirits Research Center, Mercian Corporation, 9-1 Johnan
4-chome, Fujisawa 251, Japan
* The Institute of Enology and Viticulture, Yamanashi University, Kofu
400, Japan
Received March 19, 1997
@The presence of resveratrol (3,5,4'-trihydroxystilbene) and its -glucoside,
piceid (resveratrol-3--D-glucopyranoside), together with their isomers
in wine appears to be one of the beneficial factors conferring a protective
effect against cardiovascular disease through red wine ingestion. A total
of 42 red and white wines was collected in arears from Hokkaido to Kyushu
in Japan. The wines were fractionated with a C18 Sep-pak cartridge, and
the active principles were eluted with ethyl acetate. Crude trans- and
cis-piceid were extracted from a Chinese medicine, 'Kojohkon' (Polygonum
cuspidatum), and their retention times and UV absorption were confirmed
by HPLC. trans- and cis-Resveratrol, and trans- and cis-piceid were analyzed
in a short C18 HPLC column, and cis-resveratrol was quantified from the
amount of cis-isomer converted from authentic trans-resveratrol that had
been treated by UV irradiation. The content of piceid is shown as the resveratrol
equivalent. The average content of total stilbene compounds was 4.37 mg/liter
in red wines, while only 0.68 mg/liter in white wines. Red wines made from
Pinot noir, Merlot, and Zweigeltrebe grapes all had a high resveratrol
content.
Key words: resveratrol; piceid; stilbene compounds; Japanese grape wine;
French paradox
-4-
Specific Interaction of Cytokinins and Their Analogs with
Rotenone-sensitive Internal NADH Dehydrogenase in Potato Tuber Mitochondria
Masayuki Sue, Hideto Miyoshi, and Hajime Iwamura
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Kyoto 606-01, Japan
Received March 24, 1997
@Effects of cytokinins were studied on rotenone-sensitive NADH dehydrogenase
in mitochondria from fresh potato tubers (Solanum tuberosum), in consideration
of the operation of external and rotenone-insensitive internal NADH dehydrogenases
that has not been fully accounted for in previous studies. In submitochondrial
particles (smp), zeatin was only weakly active, and zeatin riboside (ZR)
was inactive. Inhibition rates at 400 M of isopentenyladenine (iP) and
isopentenyladenosine (iPA) were 45% and 30%, respectively, and that of
BA (BA) was 64%. In intact mitochondria, the inhibition by iP and BA significantly
increased, I(/)50 being 50 and 250 M, respectively, but that by zeatin
and iPA decreased. A structure-activity study showed that hydrophobic and
steric factors are important for the activity. Cytokinins inhibited the
electron flow via natural quinone more strongly than that via synthetic
quinone. These results suggest that among the cytokinins the species that
can regulate the electron transport is iP rather than its riboside or zeatin.
Key words: cytokinins; plant mitochondria; NADH dehydrogenase; respiration;
potato tuber
-5-
Different Responses of Apolipoprotein A-I, A-IV, and B
Gene Expression during Intestinal Adaptation to a Massive Small Bowel Resection
in Rats
Kei Sonoyama and Yoritaka Aoyama
Laboratory of Food Biochemistry, Department of Bioscience and Chemistry, Faculty of Agriculture, Hokkaido University, Sapporo 060, Japan
Received April 7, 1997
@Gene expression of apolipoproteins (apo) A-I, A-IV, and B, the predominant
protein components of chylomicrons, was investigated in the residual ileum
after a massive small bowel resection in rats. A Northern blot analysis
showed that the apo A-IV mRNA level, but not the apo A-I and B mRNA levels,
in the ileum was significantly higher in the resected rats than in the
sham-operated rats 24 h and 2 wk post-surgery. RT-PCR coupled with a primer
extension assay revealed that the apo B-48 mRNA/apo B-100 mRNA ratio, i.e.,
apo B mRNA editing, in the ielum was unchanged by the resection. It is
thus concluded that, among the major intestinal apolipoproteins, apo A-IV
is the only one whose gene expression is influenced by loss of the proximal
intestine.
Key words: apolipoprotein; apo B mRNA editing; intestine; intestinal resection;
rat
-6-
Metabolic Behavior of Angiotensins in Normotensive Human
Plasma in the Supine and Upright Postures
Toshiro Matsui, Hiroshi Matsufuji, Kei Tamaya, Terukazu Kawasaki,* and Yutaka Osajima
Department of Food Science and Technology, Faculty of Agriculture, Kyushu
University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812, Japan
* Institute of Health Science, Kyushu University, 6-1 Kasugakouen, Kasuga,
Fukuoka 816, Japan
Received April 7, 1997
@The effect of activating the renin-angiotensin system on the metabolism
of angiotensins (ANGs) in normotensive human plasma was investigated. In
normotensive supine human plasma, four peptides with in vitro angiotensin
I-converting enzyme (ACE) inhibitory activity which correspond to the sequence
of ANG (3-8), ANG (4-8), ANG (5-8), and ANG (3-4) existed at a concentration
of >39 fmol/ml of plasma. When activating the renin activity by keeping
upright in posture for 60 min, ANG II and the four peptides significantly
increased as compared with the levels in the supine posture, except for
ANG I. In particular, Val-Tyr corresponding to ANG (3-4) in the upright
posture was about 4-fold more than the value in the supine posture, and
was predominantly present (447 fmol/ml of plasma) as well as ANG I. As
a result of in vitro degradation tests on ANGs, ANG (3-4) was produced
from ANG I, and not from ANG II, III or (3-8), during the 30-min incubation.
Key words: angiotensins; R-A system; metabolism
-7-
Involvements of Trp23 in the Chitin-binding and of Trp131
in the Chitinase Activity of Rye Seed Chitinase-a
Takeshi Yamagami and Gunki Funatsu
Laboratory of Protein Chemistry & Engineering, Faculty of Agriculture, Kyushu University, Fukuoka 812-81, Japan
Received April 15, 1997
@The oxidation of the tryptophan residues of rye seed chitinase-a (RSC-a)
and its isolated catalytic (Cat) domain by N-bromosuccinimide (NBS) was
investigated in the absence and presence of oligomers of N-acetylglucosamine
(GlcNAc)n. Based on the reactivity toward NBS at pH 5.9, the seven tryptophan
residues present in RSC-a are grouped into highly reactive (HR-), low reactive
(LR-), and unreactive residues. Analyses of the peptides from 1 tryptophan-
and 3 tryptophan-oxidized RSC-a showed that the HR-residue is Trp23 and
the LR-residues are Trp131 and Trp141.
@The chitin-binding ability of RSC-a was lost upon the NBS oxidation of
Trp23 at pH 5.9 or pH 7.0. This oxidation was prevented by (GlcNAc)3, which
induced a high UV-difference spectrum with maxima at 284 and 293 nm. On
the other hand, the chitinase activity of the Cat domain was greatly reduced
by the NBS oxidation of Trp131 and Trp141 at pH 5.9, while in the NBS oxidation
at pH 6.4, approximately one tryptophan residue was oxidized and about
half of the activity was retained. The NBS oxidation of the isolated Cat
domain at pH 5.9 was protected by (GlcNAc)4, which induced a UV-difference
spectrum with maxima at 284 nm and 293 nm as well as a small trough around
300 nm, similar to that observed in RSC-c. From these results and the previous
result that Trp72 in RSC-c is involved in the substrate-binding, it was
suggested that Trp23 is highly exposed on the surface of the RSC-a molecule
and involved in the chitin-binding, while Trp131 is involved in substrate-binding
in its enzyme action.
Key words: chitinase; chitin-binding protein; rye seeds; N-bromosuccinimide
oxidation; active site of enzyme
-8-
Water Permeability of Plasma Membranes of Cultured Rice,
Grape, and CH27 Cells Measured Dielectrically
Eisuke Ishikawa, Osato Miyawaki, and Kozo Nakamura
Department of Applied Biological Chemistry, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan
Received April 25, 1997
@The capacitance of suspensions of cultured rice cells (Oryza sativa L.
ssp. japonica), grape cells (Vitis sp.), and CH27 cells originated from
murine B-cell lymphoma was measured in the frequency range of 0.2 to 10
MHz. The relationship between the increase in capacitance caused by the
presence of cells at 0.4 MHz, C, and the cell density was linear.
@Measurement of capacitance was useful in measurement of transitional
changes in cell volume under external osmotic stress when sucrose was added.
From the course of volume changes with such stress, the water permeabilities
of the plasma membrane, Lp, were measured to be 0.015, 0.020, and 0.090
pm/(sEPa) at 25C, for rice cells, grape cells, and CH27 cells, respectively.
The smaller Lp for plant cells seemed to explain why preservation of plant
cells by freezing is more difficult than for animal cells. From the temperature
dependence of Lp, the apparent activation energies were calculated to be
12.0}2.9 and 13.0}5.2 kcal/mol for rice cells and CH27 cells, respectively.
Key words: freeze-induced dehydration; freezing preservation of cells;
permittivity of cell suspension; water permeability; plasma membrane
-9-
Dielectric Relaxation of Aqueous Solution with Low-molecular-weight
Nonelectrolytes and Its Relationship with Solution Structure
Akiko Saito, Osato Miyawaki, and Kozo Nakamura
Department of Applied Biological Chemistry, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan
Received June 13, 1997
@Dielectric relaxation of water molecules was measured in the frequency
range from 0.2 to 20 GHz for aqueous solutions of urea, formamide, alcohols,
and saccharides. The relaxation behavior was described well by the Debye
equation with a single relaxation process in most cases. The static permittivity
of solution and relaxation time of water in solution changed linearly with
solute concentration. The relaxation time shift of water molecules through
the existence of solute was correlated well to the first virial coefficient
of the activity coefficient of water suggesting the close relationship
between dielectric relaxation time and aqueous solution structure.
Key words: dielectric relaxation; permittivity; water structure; activity
coefficient of water; virial coefficient
-10-
A Model System for Convenient Fluorescent Labeling of
Sugar Chain in Taka-amylase A
Mikihiko Kobayashi, Yutaka Sasaki, and Yogo Chiba
National Food Research Institute, Kannondai, Tsukuba 305, Japan
Received April 25, 1997
@A convenient detection of sugar chains in Taka-amylase A (TAA) was done
by using 40 g of enzyme, where a decrease in the UV absorption of NaIO4
during the periodate oxidation reaction was monitored. The periodate-oxidized
sugar chain was labeled with a fluorescent reagent, N-1-ethylenediaminonaphthalene
(EDAN), by incubation at pH 9.5 and 30C for 1 h. The excess EDAN was
removed by either quenching with o-phthaladehyde or Bio-Gel P-2 gel adsorption.
Among the peptide fragments prepared from the EDAN-labeled TAA, a fluorescent
peptide corresponding to the sugar chain was distinguished by the ODS column.
These results suggest that periodate oxidation and subsequent fluorescent
labeling were useful for the sensitive analysis of various glycoprotein
samples.
Key words: N-1-ethylenediaminonaphthalene; fluorescent labeling; periodate
oxidation; sugar chain; Taka-amylase A
-11-
Isolation of Taka-amylase A Peptides Required for Substrate
Binding
Yutaka Sasaki, Hidenari Takahara, and Mikihiko Kobayashi*,
Department of Resource Biology, Faculty of Agriculture, Ibaraki University,
Ami, Ibaraki 300-03, Japan
* National Food Research Institute, Kannondai, Tsukuba 305, Japan
Received May 29, 1997
@An -amylase from Aspergillus oryzae, Taka-amylase A (TAA), was cleaved
into peptide fragments by an acid protease. Inactivation of TAA was greatly
retarded by the addition of -cyclodextrin or Ca2+. TAA peptide fragments
were separated into two groups having no and high affinity to the substrate,
soluble starch. This separation was done by the forced affinity chromatography
method by a column of epichlorohydrin cross-linked soluble starch gel.
Three peptides were isolated from the high-affinity fragments, purified
by the ODS-120T column, and their amino acids were sequenced. Peptides
I, II, and III originated from 2-helix, 3-helix, and 2-sheet, respectively,
and all of these were located in the (/)8 barrel of the main domain
of TAA molecule. A stereo graphic view showed that Peptides I-III were
at the cleft near the catalytic site. Occurrence of a Trp residue in all
three peptides strongly suggested that Trp was very important in the binding
of TAA to the substrate, soluble starch.
Key words: -amylase; forced affinity chromatography; substrate binding
site; Taka-amylase A
-12-
Growth Factors for a Primary Chick Muscle Cell Culture
from Shochu Distillery By-products
Luthfi D. Mahfudz, Kazuki Nakashima, Akira Ohtsuka, and Kunioki Hayashi
Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, Korimoto 1-21-24, Kagoshima 890, Japan
Received April 28, 1997
@An unidentified growth factor (UGF) was separated from shochu distillery
by-products (SDBP) and its effect on the growth of a primary chick muscle
cell culture was investigated. Chick muscle cells were isolated from fertile
eggs (13-day-old embryos) of commercial broilers. UGF was separated on
Sephadex LH-20 with a solvent system of water-methanol-ethylene dichloride
(10 : 90 : 20, v/v), and the fraction eluted between 136 and 164 min was
collected (fraction I). Fraction I was further purified by HPLC with an
Inertsil ODS-2 column using a solvent system of methanol-butanol (80 :
20, v/v). Three fractions having retention times of 3.76, 4.57, and 5.12
min were collected and are referred to as fraction A, B, and C, respectively.
In experiment 1, chick muscle cells were cultured in an m-199 medium containing
0.001, 0.01, or 0.1% of fraction I. In experiment 2, chick muscle cells
were cultured with 0.01 or 0.005% of each fraction A, B, and C. Creatine
kinase (CK) activity, protein and DNA contents were measured as indices
of myotube growth, cell growth and cell proliferation, respectively. N
-methylhistidine (N -MH) release from the muscle cell was also measured
to observe the effect on proteolysis. In experiment 1, the protein content
was significantly ( p<0.05) increased by fraction I, despite the low
dose level. CK activity was significantly ( p<0.05) higher than the
control when 0.001% of fraction I was added to the medium. However, increasing
the level beyond 0.01% did not further increase the CK activity. The DNA
content was not significantly changed. In experiment 2, the protein content,
CK activity, and DNA content were significantly ( p<0.05) higher when
fractions A and B were added to the medium. However, this was not the case
when fraction C was added. N -MH release was significantly ( p<0.05)
higher when fraction A was added, but, was significantly ( p<0.05) lower
when fraction B was added, while fraction C had no effect on N -MH release.
The present results show that SDBP contained two growth-promoting factors
for a primary chick muscle cell culture, although their modes of action
may be different.
Key words: muscle cell culture; unidentified growth factor; distillery
by-product
-13-
Nigrosporins A and B, New Phytotoxic and Antibacterial
Metabolites Produced by a Fungus Nigrospora oryzae
Masayasu Tanaka, Toshiro Fukushima, Yasuko Tsujino, and Takane Fujimori Plant Protection Research Laboratory, Japan Tobacco Inc., 6-2 Umegaoka, Aoba-ku, Yokohama, Kanagawa 227, Japan
Received April 30, 1997
@Nigrosporin A and B, two new phytotoxic and antibacterial metabolites
were isolated from a culture filtrate of Nigrospora oryzae.
@The active principles were absorbed on XAD-2 resin and purified by suceessive
ODS-HPLC. The structures were identified by spectroscopic and derivatization
analysis as naphthoquinone derivatives. The substances showed phytotoxic
activities, such as root elongation inhibition, necrotic effects, oxygen
evolution inhibition, starch synthesis inhibition, and CO2 fixation inhibition
at concentrations of 10-100 ppm. They also showed growth inhibition activity
against Bacillus subtilis in a disc diffusion assay as well as when compared
with streptomycin.
Key words: Nigrospora oryzae; phytotoxic activity; photosynthesis inhibition
activity; antibacterial activity; anthraquinone derivative
-14-
Cloning and Sequence Analysis of the Gene (alyII ) Coding
for an Alginate Lyase of Pseudomonas sp. OS-ALG-9
Jintana Kraiwattanapong, Toshihiko Ooi, and Shinichi Kinoshita
Applied Biochemistry, Department of Molecular Chemistry, Faculty of Engineering, Hokkaido Graduate University, Sapporo, Hokkaido 060, Japan
Received May 6, 1997
@Pseudomonas sp. OS-ALG-9 produces several kinds of alginate-degrading
enzymes both intra- and extracellularly. As a second alginate lyase of
this bacterium, the gene encoding alyII has been cloned in Escherichia
coli
JM109 by shotgun techniques and then sequenced. The alyII gene has an open
reading frame of 2141 bp encoding 713 amino acid residues with a calculated
molecular mass of 79,803 Da. The deduced amino acid sequence did not show
any extensive similarity with those of other known alginate lyases, however,
hydrophobic cluster analysis showed that alyII belonged to class 3 of alginate
lyases. The alginate lyase from E. coli harboring the alyII gene showed
a single active band, which coincided with one of four major alginate lyases
from the crude cell extracts of Pseudomonas sp. OS-ALG-9 on a zymogram.
Key words: alginate; alginate lyase gene; Pseudomonas sp.; gene cloning
-15-
Secretion of Human Interleukin-2 in Biologically Active
Form by Bacillus brevis Directly into Cultute Medium
Yasushi Takimura, Masashi Kato, Toshihiro Ohta,* Hideo Yamagata,* and Shigezo Udaka**,
Department of Applied Biological Sciences, Faculty of Agriculture, Nagoya
University, Nagoya 464, Japan
* School of Life Science, Tokyo University of Pharmacy and Life Science,
1432-1 Horinouchi, Hachioji, Tokyo 192-03, Japan
** Department of Brewing and Fermentation, Tokyo University of Agriculture,
Sakuragaoka, Setagaya, Tokyo 156, Japan
Received May 8, 1997
@We constructed an efficient system for synthesis and secretion of human
interleukin-2 (IL-2) by Bacillus brevis. The secretion vector we constructed
had strong promoters and contained the region coding for the signal peptide
of the gene for B. brevis 47 cell-wall protein, followed directly by the
gene encoding mature IL-2. Modification of the signal peptide and use of
a protease-deficient mutant of B. brevis HPD31 increased productivity.
When the signal peptide was more basic near its amino terminal and more
hydrophobic in the middle region, IL-2 production increased 20 fold. Production
by the mutant harboring the secretion vector was four fold that of the
parent harboring the same plasmid. The yield of IL-2 increased further
to 0.12 g/liter, when cultural conditions were made optimal, such by the
addition of Tween 40 to the medium. The IL-2 produced by B. brevis had
the same biological activity as authentic IL-2. Biologically active human
IL-2 was produced efficiently and secreted directly into the medium by
B. brevis.
Key words: interleukin-2; Bacillus brevis; protein secretion; signal peptide
-16-
Degree of Polymerization of Cellulose from Acetobacter
xylinum BPR2001 Decreased by Cellulase Produced by the Strain
Naoki Tahara, Mari Tabuchi, Kunihiko Watanabe, Hisato Yano, Yasushi Morinaga, and Fumihiro Yoshinaga
Bio-Polymer Research Co., Ltd., KSP R&D Business-Park Bldg., 3-2-1, Sakato, Takatsu-ku, Kawasaki 213, Japan
Received May 8, 1997
@Acetobacter xylinum produces both cellulase and bacterial cellulose,
but some report believed that this cellulase activity does not decrease
the degree of polymerization (DP) of bacterial cellulose during cultivation.
A. xylinum subsp. sucrofermentans BPR2001 produces two enzymes that hydrolyze
CM-cellulose and cellotriose, respectively. We examined the effect of the
two cellulase activities on the DP of bacterial cellulose when bacterial
cells were cultured with agitation at pH 4, where little cellulase is produced,
and at pH 5, where much cellulase is produced. The weight-average degree
of polymerization (DPw) of bacterial cellulose remained in the range of
14,000 of 16,000 during cultivation at pH 4, but at pH 5, the DPw decreased
from 16,800 to 11,000. The mechanical strength of a sheet prepared from
the bacterial cellulose produced at pH 4 was higher than those of BC produced
at pH 5. These results suggest that the two cellulase activities cause
the decrease in DP and deterioration of physical properties of bacterial
cellulose seen during cultivation.
Key words: bacterial cellulose; cellulase; degree of polymerization; Acetobacter
-17-
Structural Analysis of N-Glycans of Storage Glycoproteins
in Soybean (Glycine max. L) Seed
Yoshinobu Kimura, Akira Ohno, and Shigeaki Takagi
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Tsushima-Naka 1-1-1, Okayama 700, Japan
Received May 8, 1997
@The structures of N-linked sugar chains (N-glycans) of storage glycoproteins
in soybean seeds have been identified. Eight pyridylaminated (PA-) N-linked
sugar chains were derived and purified from hydrazinolysates of the storage
glycoproteins by reverse-phase HPLC and size-fractionation HPLC. The structures
of the PA-sugar chains purified were first identified by two-dimensional
PA-sugar chain mapping and ion-spray mass analysis, considering the results
of sugar composition analysis or sequential exoglycosidase digestion. The
deduced structures were further analyzed by ion-spray tandem mass spectrometry
and 500 MHz 1H-NMR spectrometry. The eight structures fell into two categories;
the major class (96.6%) was a typical high mannose-type, the minor class
was a xylose containing-type (Man3Xyl1GlcNAc2, Man3Fuc1Xyl1GlcNAc2; 3.4%).
Key words: N-glycan; plant glycoprotein; allergenic oligosaccharide; ion-spray
mass spectrometry; Glycine max
-18-
Synthesis, Biological Activity, and Metabolism of 8',8',8'-Trideuteroabscisic
Acid
Yasushi Todoroki, Sei-ichi Nakano, Nobuhiro Hirai, Toshiaki Mitsui,* and Hajime Ohigashi
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto
University, Kitashirakawa-oiwake-cho, Sakyo-ku, Kyoto 606-01, Japan
* Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata
University, 2-8050 Ikarashi, Niigata 950-21, Japan
Received May 12, 1997
@An 8',8',8'-trideuterated analog of abscisic acid (ABA) was diastereoselectively
synthesized as a new analog of ABA that is resistant to 8'-hydroxylation,
the first metabolic reaction of ABA, owing to the primary kinetic isotope
effect. (+)-8',8',8'-Trideutero-ABA showed long-term activity in the rice
elongation assay. The rate of metabolism of this analog in rice cell suspension
culture was about two fold slower than that of (+)-ABA. The concentration
of 8',8'-dideuterophaseic acid produced was about 1/3 that of phaseic acid
converted from (+)-ABA. This result indicated that the long-lasting activity
of the (+)-trideutero-ABA in the rice assay was the result of the delayed
8'-hydroxylation as expected.
Key words: abscisic acid; 8',8',8'-trideuteroabscisic acid; phaseic acid;
primary kinetic isotope effect; rice cell suspension culture
-19-
Hyperproduction of L-Threonine by an Escherichia coli
Mutant with Impaired L- Threonine Uptake
Kazuyuki Okamoto, Kuniki Kino, and Masato Ikeda
Technical Research Laboratories, Hofu Plant, Kyowa Hakko Kogyo Co., Ltd., 1-1 Kyowa-machi, Hofu, Yamaguchi 747, Japan
Received May 14, 1997
@An efficient production strain for L-threonine fermentation was derived
from Escherichia coli by multiple rounds of mutation programs that aimed
at deregulation of the L-threonine biosynthetic pathway and blocking of
L-threonine degradation pathways. When the optimum amount of DL-methionine
was added, this strain KY10935, an L-methionine auxotroph, gave 100 g/liter
L-threonine after 77 h cultivation. In this strain, key enzymes in the
L-threonine biosynthetic pathway were highly derepressed, but some were
inhibited by lower concentrations of L-threonine than the accumulated level.
Such incomplete deregulation of the pathway was accounted for by the intracellular
concentration of L-threonine being lower than the extracellular level.
In an assessment of L-threonine transport in terms of phenotypic growth
responses to the amino acid, L-threonine-auxotrophic mutants with a lesion
in the L-threonine operon were derived from strain KY10935 by selection
for auxotrophy for dipeptide L-alanyl-L-threonine or glycyl-L-threonine,
the transport systems of which were different from those of L-threonine.
All three independent mutants isolated needed an extraordinarily high concentration
(10 mg/ml) of L-threonine, but grew in the presence of a low concentration
(10 g/ml) of either dipeptide, indicating that strain KY10935 had impaired
L-threonine uptake. These results suggested that the strain had an unusual
mechanism of L-threonine hyperproduction: the inability to take up L-threonine
that had accumulated extracellularly decreased the steady-state level of
intracellular L-threonine, freeing the remaining regulatory steps of feedback
inhibition.
Key words: threonine production; transport mutant of Escherichia coli ;
threonine uptake; strain improvement
-20-
Activation of Macrophages by Sulfated Glycopeptides in
Ovomucin, Yolk Membrane, and Chalazae in Chicken Eggs
Hiroko Tanizaki,* Hiroki Tanaka,* Hiroyuki Iwata,** and Akio Kato*,
* Department of Biological Chemistry, and ** Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753, Japan
Received May 15, 1997
@Sulfated glycopeptides in ovomucin, chalazae and yolk membrane were found
to activate cultured macrophage-like cells, J774.1, and TGC-induced macrophages
from the peritoneal cavity of male mice. The macrophage-stimulating activity
was estimated by the growth and morphology of the cells, H2O2 generation,
and interleukin-1 (IL-1) production from the cells. The in vitro culture
assay with macrophages showed that the protease digests of ovomucin, yolk
membrane, and chalazae induced morphologic alteration and increased H2O2
generation and IL-1 production in lower concentration (100 g/ml). The
isolation of the components having macrophage-stimulating activity was
attempted to elucidate the molecular mechanism. The O-linked carbohydrate
chains, consisting of N-acetylgalactosamine, galactose, N-acetylneuraminic
acid and sulfate, in the sulfated glycopeptide were identified as a component
having macrophage-stimulating activity.
Key words: sulfated glycopeptide; ovomucin; yolk membrane; chalazae; macrophage
activation
-21-
Purification and Characterization of Cystine Lyase a from
Broccoli Inflorescence
Koji Ukai and Jiro Sekiya
Division of Applied Life Sciences, Graduate School of Agricultural Sciences, Kyoto University, Kyoto 606-01, Japan
Received May 15, 1997
@One of the three isoforms of an enzyme degrading L-cystine was purified
to homogeneity from broccoli (Brassica oleracea var. italica) inflorescences,
with use of a sensitive assay based on derivatization of a reaction product
with monobromobimane. The reaction product with a thiol group was found
to be thiocysteine from results of liquid chromatography-mass spectrometry
and high-resolution mass spectrometry. Pyruvate was also a reaction product,
formed in equimolar amounts. The purified enzyme catalyzed -elimination
of L-cystine to yield thiocysteine, pyruvate and possibly ammonia, so it
was cystine lyase a. L-Cystine but not D-cystine was a substrate of the
enzyme. S-Methyl L-cysteine sulfoxide and S-ethyl L-cysteine sulfoxide
were substrates but were less suitable than L-cystine. L- and D-cysteine
and also cystathionine were not substrates. The purified enzyme (Mr 186,000)
was composed of four identical subunits (Mr 45,000) and was pyridoxal 5'-phosphate-dependent.
Key words: Brassica oleracea var. italica; broccoli; cystine lyase; thiocysteine;
purification
-22-
Effects of Plant Growth Regulators on Shoot Growth and
Flowering of a Perennial Paddy Weed, Sagittaria pygmaea Miq.
Takumi Yoshimura, Hitoshi Kuramochi,* Makoto Konnai,* Hideharu Seto,** Takeshi Sassa,*** and Koichi Yoneyama*,
KEI Chemical Research Institute Co., Ltd., Shioshinden Fukude-cho,
Iwata-gun, Shizuoka 437-12, Japan
* Weed Science Center, Utsunomiya University, 350 Mine, Utsunomiya 321,
Japan
** The Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa,
Wako 351-01, Japan
*** Department of Bioproduction, Faculty of Agriculture, Yamagata University,
1-23 Wakaba, Tsuruoka 997, Japan
Received May 22, 1997
@Plant growth regulators (PGRs) including gibberellins (GAs) were examined
for their effects on shoot growth and flowering of a perennial paddy weed,
Sagittaria pygmaea Miq. Among PGRs tested, only GAs (A1, A3, A4, and A5),
AC-94377 [1-(4-chloro-1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)cyclohexanecarboxamide]
and 2,6-diisopropylphenoxyacetic acid (DIPA) promoted both shoot growth
and flowering. Structural requirements of GAs for promotion of flowering
seemed to be different from those for shoot growth of the weed. In addition,
since the course of flowering induced by DIPA was clearly different from
that observed in plots treated with GA3 or AC-94377, different mechanisms
may be involved in promotion of flowering.
Key words: 2,6-diisopropylphenoxyacetic acid; flowering; gibberellin; plant
growth regulator; Sagittaria pygmaea Miq.
-23-
Protective Effect of Green Tea Extract and Tea Polyphenols
against the Cytotoxicity of 1,4-Naphthoquinone in Isolated Rat Hepatocytes
Chika Miyagawa, Chen Wu, David Opare Kennedy, Teruyo Nakatani, Kimiko Ohtani, Senji Sakanaka,* Mujo Kim,* and Isao Matsui-Yuasa
Department of Food and Nutrition, Faculty of Human Life Science, Osaka
City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558, Japan
* Central Research Laboratories of Taiyo Kagaku Co., Ltd., Yokkaichi, Mie
510, Japan
Received May 22, 1997
@The cytoprotective effect of green tea extract and its phenolic compounds
against 1,4-naphthoquinone-induced hepatotoxicity was evaluated in primary
cultured rat hepatocytes. After exposure to 1,4-naphthoquinone, lactate
dehydrogenase (LDH) leakage and cell viability were both improved by the
presence of the tea extract and tea polyphenols. This cytoprotective effect
was related to the structure of tea polyphenols, the galloyl group of (-)-epigallocatechin-3-gallate
and (-)-epicatechin-3-gallate being particularly effective. The production
of liquid peroxidation by 1,4-naphthoquinone was not inhibited by the tea
extract nor by tea polyphenol addition. After 2 h of incubation, the protein
thiol concentration was reduced by 1,4-naphthoquinone, but this reduction
was prevented by the tea extract and tea polyphenols. The reduction in
protein thiol content of the cells closely paralleled the LDH leakage and
loss of cell viability. These results suggest that the mechanism of protection
by tea polyphenols against 1,4-naphthoquinone-induced toxicity to rat hepatocytes
was due to the maintenance of protein thiol levels.
Key words: green tea extract; tea polyphenols; 1,4-naphthoquinone; protein-SH;
hepatocytes
-24-
(E )-2-(4'-Methyl-3'-pentenylidene)-4-butanolide, Named
-Acariolide: A New Monoterpene Lactone from the Mold Mite, Tyrophagus
putrescentiae (Acarina: Acaridae)
Atsushi Morino, Yasumasa Kuwahara,*, Sigeru Matsuyama, and Takahisa Suzuki
Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki
305, Japan
* Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto
University, Sakyo-ku, Kyoto 606-01, Japan
Received May 26, 1997
@Reinvestigation of the opisthonotal gland secretion of the mold mite,
Tyrophagus putrescentiae, resulted in the isolation of a new monoterpene
lactone, whose chemical structure was elucidated as (E )-2-(4'-methyl-3'-pentenylidene)-4-butanolide
(3), to which we gave the trivial name -acariolide in relation to -acaridial
{1, (E )-2-(4-methyl-3-pentenylidene)-butanedial}. The compound was synthesized
by LiAlH3 (OEt) reduction of 1 and subsequent oxidation involving simultaneous
cyclization by using Ag2CO3 on Celite. Both the E- and Z-isomers of -acariolide
(3 and 4) were also prepared by the reaction of -ethoxaly--butyrolactone
(6) and 4-methyl-3-pentenal under basic conditions. Their NMR spectra were
compared with each other, and the geometry of the pentenylidene double
bond of the isolated compound was concluded as being E.
Key words: (E )-2-(4'-methyl-3'-pentenylidene)-4-butanolide; -acariolide;
Tyrophagus putrescentiae; opisthonotal gland; Acaridae
-25-
Immunopotentiating Activity of the Water-soluble Lignin
Rich Fraction Prepared from LEM-The Extract of the Solid Culture Medium
of Lentinus edodes Mycelia-
Yoshiki Yamamoto, Hiroyuki Shirono, Keiko Kono, and Yasuhiro Ohashi*
Biochemistry Research Laboratories, JCR Pharmaceuticals Co., Ltd., 2-2-10
Murotani, Nishi-ku, Kobe 651-22, Japan
* Iizuka Institute, Noda Shokukin Kogyo Co., Ltd., 295 Nanakodai, Noda,
Chiba 278, Japan
Received May 29, 1997
@The water-soluble lignin in LEM (the extract of the solid culture medium
of Lentinus edodes mycelia) has been known to have antiviral and immunopotentiating
activities in vivo and in vitro. The water-soluble lignin rich fraction
(JLS-18) was prepared from LEM using ultrafiltration and hydrophobic column
chromatography. JLS-18 showed about 70 times higher antiviral activity
than LEM in vitro. JLS-18 activated the cytotoxicity of NK cells and macrophages,
and activated T cells in vitro. JLS-18 also induced interleukin 6 (IL-6)
secretion from human leukocytes infected with Sendai virus in vitro. These
data showed that JLS-18, the water-soluble rich fraction of LEM, had antiviral
and immunopotentiating activities.
Key words: JLS-18; water-soluble lignin; LEM; immunopotentiating activity
-26-
Enzymatic Properties of Double Mutant Enzymes at Asp51
and Trp49 and Asp51 and Tyr57 of RNase Rh from Rhizopus niveus
Kazuko Ohgi, Mitsuaki Takeuchi, Masanori Iwama, and Masachika Irie
Department of Microbiology, Hoshi College of Pharmacy, Ebara 2-4-41, Shinagawa-ku, Tokyo 142, Japan
Received June 3, 1997
@Mutation of Asp51 of a base-nonspecific RNase, RNase Rh, to Ser, Thr,
or Gln makes the enzyme more preferential for the dinucleoside phosphate
(XpY) having G and C at the 5'-side (X). On the other hand the mutation
of one of the B1 site components, Tyr57 to Trp, and Trp49 to Phe makes
the enzyme more preferential for purine bases and pyrimidine bases, respectively.
In this study, to obtain more specific RNases and RNases with different
base specificity, we prepared double-mutant enzymes that have Ser, Thr,
and Asn at the 51st position and Trp at the 57th position or Phe at the
49th position, and their enzymatic specificities were studied with XpYs
as substrates. The double-mutant enzymes D51SY57W and D51TY57W are more
guanylic acid preferential than the mother single-mutant enzymes, D51S
and D51T, respectively. They are extremely guanylic preferential RNases.
D51NY57W is more a guanylic acid preferential enzyme than D51N, but cytidylic
acid preference is of a similar order to that of D51N. The double mutant
enzymes D51NW49F and D51TW49F showed an increased cytidylic acid preference
as well as guanylic acid preference as compared to the mother single-mutant
enzymes, D51T and D51N. The results of analysis of base specificity by
the release of mononucleotides from RNA and the rates of hydrolysis of
homopolynucleotides led to the same conclusion as in the case of the hydrolysis
of XpY.
Key words: base non-specific RNase; base specificity; Rhizopus niveus;
site-directed mutagenesis
-27-
Structural Property and in Vitro Self-assembly of Shark
Type I Collagen
Yoshihiro Nomura, Masaya Yamano, Chikayuki Hayakawa, Yasuhiro Ishii, and Kunio Shirai
Applied Protein Chemistry, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183, Japan
Received June 9, 1997
@The main structure of shark type I collagen is similar to that of land
mammals, with a partial difference in amino acid sequence and post-translational
modification. By static light scattering, the weight-average molecular
weight of shark collagen (7.52~105) suggests the presence of some aggregated
molecules, oligomeric collagen. The self-assembly curve of shark collagen
had a shorter lag phase and a longer growth phase than that of pig collagen.
The optimum temperature and pH of shark collagen self-assembly is different
from that of pig collagen.
Key words: type I collagen; shark; self-assembly; static light scattering;
electrophoretic light scattering
-28-
Note
Growth Suppressing Activity for Endothelial Cells Induced from Macrophages
by Carboxymethylated Curdlan
Shigeyuki Usui, Toshiyuki Matsunaga, Shigeo Ukai, and Tadashi Kiho*
Department of Hygienic Chemistry, Gifu Pharmaceutical University, 5-6-1 Mitahora-higashi, Gifu 502, Japan
Received April 7, 1997
@A carboxymethylated derivative of a linear (13)--D-glucan (CMCD)
from Alcaligenes faecalis var. myxogenes acted directly on mouse peritoneal
macrophages and mouse lymphoma P388D1 cells, and induced a growth suppressing
activity for bovine artery endothelial cells (BAEs) from themselves at
a concentration of 100 g/ml. The suppressing activity was also detected
in the mouse serum administered as an i.p. injection of CMCD at a dose
of 100 mg/kg, suggesting that the growth suppressing activity was induced
from macrophages potentiated by CMCD in vivo.
Key words: curdlan; endothelial cell; macrophage; cytotoxicity; growth
inhibition
-29-
Note
Lipase-catalyzed Synthesis of Arbutin Cinnamate in an Organic Solvent and
Application of Transesterification to Stabilize Plant Pigments
Nobuyoshi Nakajima, Kohji Ishihara,* Shingo Matsumura,**, Hiroki Hamada,** Kaoru Nakamura,*** and Tsutomu Furuya**
Department of Nutritional Science, Okayama Prefectural University, Soja,
Okayama 719-11, Japan
* Department of Chemistry, Kyoto University of Education, Fushimi-ku, Kyoto
612, Japan
** Department of Applied Science, Okayama University of Science, Ridai-cho,
Okayama 700, Japan
*** Institute for Chemical Research, Kyoto University, Uji, Kyoto 611,
Japan
Received April 7, 1997
@Arbutin cinnamate was synthesized from arbutin (4-hydroxyphenyl -D-glucopyranoside)
and vinyl cinnamate by regioselective transesterification with a bacterial
lipase in acetonitrile. The product was identified by NMR and FAB-MS analyses.
These spectra showed that one ester bond was formed between the primary
alcohol moiety of the D-glucose of arbutin and the carboxyl residue of
cinnamic acid. Furthermore, plant pigments such as isoquercitrin (quercetin
3-O--D-glucopyranoside) and callistephin (pelargonidin 3-O--D-glucopyranoside)
were also converted to their corresponding cinnamate esters in the same
manner.
Key words: lipase; transesterification; arbutin; cinnamic acid; plant pigment
-30-
Note
New Acylated Anthocyanins from Brassica campestris var. chinensis
Masahiro Suzuki, Tadahiro Nagata, and Norihiko Terahara*
National Food Research Institute, Ministry of Agriculture, Forestry
and Fisheries, Tsukuba, Ibaraki 305, Japan
* Department of Food Science and Technology, College of Horticulture, Minami-Kyushu
University, Takanabe, Miyazaki 884, Japan
Received April 15, 1997
@Two new acylated anthocyanins were isolated from beninabana, Brassica
campestris var. chinensis, in addition to two known anthocyanins. The structures
were established by spectral analyses.
Key words: Brassica campestris; acylated anthocyanins; p-coumaric acid;
malonic acid; sinapic acid
-31-
Note
Manufacturing High Purity Maltose and Maltotetraose from Starch by a Novel
and Efficient Procedure Named ''Reducing End Modification Method''
Yohji Ezure, Shigeaki Maruo, Masahiko Kojima, Tomonori Sakai, Hirofumi Yamamoto, Noriyuki Tachikake, Hirotsugu Ogawa, and Masaya Toda
Discovery Research Laboratory III, Nippon Shinyaku Co., Ltd., Nishioji-Hachijo, Minami-ku, Kyoto 601, Japan
Received April 15, 1997
@A novel and efficient procedure named ''reducing end modification method''
was developed for manufacturing high purity maltooligosaccharides. By the
method, high purity maltose and maltotetraose were prepared from starch.
Starch was liquefied, debranched, and oxidized with sodium hypochlorite
at the reducing end. The reaction mixture was treated with -amylase or
maltotetraose-forming amylase from Pseudomonas stutzeri IFO-3773, resulting
in the production of high purity maltose or maltotetraose, oxidized maltooligosaccharides,
and oxidized undigested dextrin. High purity maltose (maltose content,
99.9%) or maltotetraose (maltotetraose content, 95%) was obtained very
effectively from the reaction mixture by chromatography on a strong acid
cation exchange resin (Na+).
Key words: maltose; maltotetraose; -amylase; maltotetraose-forming amylase;
reducing end modification method
-32-
Note
Heterodimeric Aminopeptidase A from Bacillus licheniformis NS115
Tae-Kwang Oh, Mi-Ja Park, Jung-Kee Lee, Hyung-Kwoun Kim, and Hee-Sop Nam*
Microbial Enzyme Research Unit, Korea Research Institute of Bioscience
& Biotechnology, P.O. Box 115, Yusung, Taejon 305-600, South Korea
* Research and Development Center, Nong Shim Co., Ltd., Kyung-Ki 430-030,
South Korea
Received April 21, 1997
@An aminopeptidase A (EC 3.4.11.7) was purified to homogeneity from Bacillus
licheniformis NS115 and its enzymatic properties were characterized. The
enzyme had an apparent molecular mass of 64 kDa, consisting of heterodimeric
42 kDa and 22 kDa subunits, and is a new enzyme from N-terminal analysis
of heavy and light subunits. The light subunit had no catalytic activity
against the substrate and apparent Km values of heavy and whole enzyme
were 0.26 and 0.087 mM of -glutamyl-p-nitroanilide, respectively. Key
words: Bacillus licheniformis; aminopeptidase A; heterodimer
-33-
Note
Stimulation of Tumor Necrosis Factor and Interleukin-1 Productivity by
the Oral Administration of Cabbage Juice to Rats
Wataru Komatsu, Kazumi Yagasaki, Yutaka Miura, and Ryuhei Funabiki Department of Applied Biological Science, Tokyo Noko University, Fuchu, Tokyo 183, Japan
Received April 30, 1997
@The effect of orally administering cabbage juice on tumor necrosis factor
(TNF) and interleukin-1 (IL-1) productivity was studied in resident
peritoneal macrophages from normal and hepatoma-bearing rats. The productivity
of TNF and IL-1 was stimulated by gastric intubation of cabbage juice in
the normal state, but not in the hepatoma-bearing state where the production
of these cytokines had already been stimulated. From these results, cabbage
may contain some effective component(s) that can be absorbed from the gastrointestinal
tract to stimulate the production of TNF and IL-1.
Key words: cabbage juice; tumor necrosis factor; interleukin-1; hepatoma
-34-
Note
An Established Hybridoma Clone Producing a Monoclonal Antibody against
Vibrio anguillarum
Koji Ikura, Shinsuke Kodama, Hiroyuki Hashimoto, Kaeko Hayashi, Hajime Mori, Masatoshi Ichida, Minoru Sorimachi,* Ken-ichi Kudo,** and Saburo Hara
Kyoto Institute of Technology, Matsugasaki, Kyoto 606, Japan
* National Research Institute of Aquaculture, Nansei, Mie 516-01, Japan
** Sanwa Cornstarch Co., Ltd., Kashihara, Nara 634, Japan
Received May 1, 1997
@Vibrio anguillarum is a pathogenic microorganism of vibriosis, an infectious
disease found in various fish species. A mouse hybridoma clone, named C5,
that produced a monoclonal antibody to V. anguillarum was established.
The specific reaction of C5 antibody with V. anguillarum was confirmed
by the pre-adsorption effect of the V. anguillarum cells in ELISA and a
cell immunoprecipitation experiment. Western blotting analysis indicated
that the C5 antibody recognized a high molecular weight substance extracted
from cells with detergents.
Key words: monoclonal antibody; Vibrio anguillarum
-35-
Note
Antioxidant Activity of Ferulic Acid -Glucuronide in the LDL Oxidation
System
Takeo Ohta, Tomoko Nakano, Yukari Egashira, and Hiroo Sanada
Department of Bioproduction Science, Faculty of Horticulture, Chiba University, 648 Matsudo, Matsudo-shi, Chiba 271, Japan
Received May 6, 1997
@Antioxidant activity of ferulic acid -glucuronide, which is prepared
from the plasma of feruloyl arabinose fed rats, in the CuSO4-induced LDL
autoxidation system was studied. It has been found that absorbed ferulic
acid occurred as the -glucuronide and that the antioxidant activity of
ferulic acid -glucuronide, which has not only the hydrophobic ferulic
acid moiety but also a hydrophilic sugar moiety, is stronger than ferulic
acid in the LDL oxidation system.
Key words: antioxidant; low density lipoprotein; cereal cell wall; ferulic
acid; glucuronide
-36-
Note
Two-mode Analysis by High-performance Liquid Chromatography of -Aminobenzoic
Ethyl Ester-derivatized Monosaccharides
Shoichi Yasuno, Takeomi Murata,*, Kazuko Kokubo,* Takashi Yamaguchi,* and Masugu Kamei
Sugiyama Chemical and Industrial Laboratory, 11 Kagetori-cho, Totsuka-ku,
Yokohama, Kanagawa 245, Japan
* Honen Corporation, 1-2-3 Ohtemachi, Chiyoda-ku, Tokyo 100, Japan
Received May 8, 1997
@-Aminobenzoic ethyl ester (ABEE)-derivatized monosaccharides were separated
by HPLC with a trifluoroacetic acid (TFA) solution or borate buffer as
the eluent. In the case of the TFA solution, ABEE-derivatized monosaccharides
of the neutral and amino sugars found in animal glycoproteins were separated
in a simultaneous analysis. In the case of the borate buffer, ABEE-derivatized
monosaccharides of identical molecular weights such as ABEE-Gal, -Glc,
and -Man were separated as stereoisomers. Glucuronic acid and galacturonic
acid were detected and separated within 8 min. The relationship between
the peak areas and the amounts of ABEE-derivatized monosaccharides was
linear in the range of 1 to 1000 pmol.
Key words: -aminobenzoic ethyl ester; monosaccharide analysis; trifluoroacetic
acid; potassium borate buffer
-37-
Note
Photocatalysis-dependent Inactivation of Lactobacillus Phage PL-1 by a
Ceramics Preparation
Yukari Kakita, Nobuhiro Kashige, Fumio Miake, and Kenji Watanabe
Faculty of Pharmaceutical Sciences, Fukuoka University, Nanakuma 8-19-1, Jonan-ku, Fukuoka 814-80, Japan
Received May 16, 1997
@A ceramics preparation (Cleansand-205), which was coated with a mixture
of the oxides of Si, Al, Ti, and Ag, was found to inactivate Lactobacillus
phage PL-1 suspended in a buffer solution. The inactivation of phage was
dependent on the amounts of Cleansand-205 added, and the reaction obeyed
almost first-order reaction kinetics. The phage inactivation was considerably
accelerated by the presence of light.
Key words: ceramics; Lactobacillus phage; phage inactivation; photocatalysis;
active oxygens
-38-
Note
Human Placental Fructose-6-phosphate,2-kinase/Fructose-2,6-bisphosphatase:
Its Isozymic Form, Expression and Characterization
Ryuzo Sakakibara, Mika Uemura, Takafumi Hirata, Noriko Okamura, and Mie Kato*
Department of Biochemistry, School of Pharmaceutical Sciences, Nagasaki
University, 1-14 Bunkyo-machi, Nagasaki, Nagasaki 852, Japan
* Department of Protozoology, Institute of Tropical Medicine, Nagasaki
University, 1-12-4 Sakamoto, Nagasaki 852, Japan
Received May 19, 1997
@The nucleotide sequence of 1981 bp cDNA containing the entire coding
region of a human placental fructose-6-phosphate,2-kinase/ fructose-2,6-bisphosphatase
was determined. The sequence encodes 469 amino acids and, based on homology
to the rat testis enzyme, appears to be the testis-type isozyme expressed
in placenta. The enzyme was expressed in Escherichia coli BL21 (DE3) by
using a T7 RNA polymerase-based expression system and purified to homogeneity.
The expressed enzyme was bifunctional with specific activities of 75 and
80 mU/mg of kinase and phosphatase, respectively. Kinetic parameters of
the expressed enzyme are similar to those of the rat testis enzyme.
Key words: bifunctional enzyme; fructose-6-phosphate,2-kinase/ fructose-2,6-bisphosphatase;
human placenta; isozyme; testis
-39-
Note
Nitric Oxide Formation by Macrophages Stimulated with Water Extracts from
Meats and Offals
Misao Miwa, Kiyohiro Shibata,* Kiyomi Nagayama,* and Katuhiro Aikawa**
National Food Research Institute, Tsukuba, Ibaraki 305, Japan
* Ibaraki University, Ami, Inasiki-gun, Ibaraki 300-03, Japan
** Chugoku National Agricultural Experiment Station, Ohda, Shimane 694,
Japan
Received May 21, 1997
@Water extracts from meats and offals were incubated with a macrophage
cell line (RAW 264.7), and nitrite in the medium was measured as an index
of the macrophage stimulating activity. Ten of 38 water extracts had macrophage
stimulants and chicken meat, chicken gizzard, cattle reticulorumen, swine
stomach, and swine cerebrum had high activities.
Key words: macrophage stimulation; nitric oxide; tumor cytotoxicity; nitrite;
meats and offals
-40-
Note
Synthesis of Asymmetrically Labeled Sucrose by a Recombinant Sucrose Synthase
Tomonori Nakai, Naoto Tonouchi,* Takayasu Tsuchida,* Hitoshi Mori,** Fukumi Sakai, and Takahisa Hayashi
Wood Research Institute, Kyoto University, Gokasho, Uji, Kyoto 611,
Japan
* Bio-Polymer Research Co., Ltd., KSP, Takatsu-ku, Kawasaki, Kanagawa 213,
Japan
** Faculty of Agriculture, Nagoya University, Higashiyama-ku, Nagoya, Aichi
464-01, Japan
Received May 22, 1997
@About 80% of radioactivity was recovered in asymmetrically labeled sucrose
from UDP-[14C]glucose or [14C]fructose with recombinant mung bean sucrose
synthase expressed in Escherichia coli harboring pEB-01. This high recovery
is due to the fact that the enzyme conserving the activity of sucrose synthase
has a similar affinity for UDP-glucose and fructose to an intact enzyme
from the mung bean, but a lower affinity for sucrose.
Key words: recombinant enzyme; sucrose; asymmetrically labeling; mung bean
-41-
Note
Biodegradabilities of Ethylenediamine-N,N '-disuccinic Acid (EDDS) and
Other Chelating Agents
Rikiya Takahashi, Naoshi Fujimoto, Masaharu Suzuki, and Takakazu Endo*
Laboratory of Environmental Science for Brewing, Department of Brewing
and Fermentation, Faculty of Agriculture, Tokyo University of Agriculture,
1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156, Japan
* Central Research Laboratory, Nitto Chemical Industry Co., Ltd., 10-1
Daikoku-cho, Tsurumi-ku, Yokohama 230, Japan
Received June 18, 1997
@Biodegradabilities of chelating agents were tested with activated sludge.
Ethylenediaminetetraacetic acid (EDTA) remained intact in the effluent
even after acclimation for 100 days, but propanediamine-N,N '-disuccinic
acid (PDDS) and nitrilotriacetic acid (NTA) were biodegraded after acclimation
for 5 and 23 days, respectively. Optical isomers of ethylenediamine-N,N
'-disuccinic acid (EDDS) had different biodegradabilities: SS- and RS-isomers
were susceptible to biodegradation, but the RR-isomer was resistant. SS-isomer
was degraded even by activated sludge without acclimation.
Key words: biodegradation; ethylenediamine-N,N '-disuccinic acid; nitrilotriacetic
acid; propanediamine-N,N '-disuccinic acid
-42-
Short Communication
Panton-Valentine Leukocidin Genes in a Phage-like Particle Isolated from
Mitomycin C-Treated Staphylococcus aureus V8 (ATCC 49775)
Jun Kaneko, Takahiro Kimura, Yoshiyuki Kawakami,* 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
* Division of Clinical Microbiology, Department of Medical Technology,
School of Allied Medical Sciences, Shinshu University, Asahi 3-1-1, Matsumoto,
Japan
Received July 24, 1997
@The staphylococcal Panton-Valentine leukocidin (PVL) genes [lukS-PV-lukF-PV]
existed in a hexagonal phage-like particle (PVL) isolated from mitomycin
C-induced Staphylococcus aureus V8 (ATCC 49775). The genome packed in PVL
was a linear double-stranded 40-kb DNA with single-stranded cohesive ends
(cos). The [lukS-PV-lukS-PV], attP, and int (integrase gene) of PVL were
all located very close to one another within a 4.0 kb-segment on the genome
in the order given, and the segment is located at the center from the left
and the right cos sites. In addition, the [lukS-PV-lukF-PV]-attP-int region
contains 5 direct repeat sequences that show high similarity with the recombinase-binding
sites of bacteriophages of S. aureus.
Key words: Staphylococcus aureus V8 (ATCC 49775); lukS-PV; LukF-PV; integrase;
temperate phage (PVL) carrying Panton-Valentine leukocidin genes