(Vol.61 No.9 1997)
|Review-
Biocatalysis in Organic Synthesis: The Use of Nitrile- and Amide-hydrolyzingMicroorganisms
Takeshi Sugai, Takahiro Yamazaki, Masahiro Yokoyama, and Hiromichi
Ohta 1419
Two Oxazolyl Compounds and a Monosubstituted ฟ-Pyrone
as Free-radical Scavengers Isolated from a Fungus
Yasujiro Morimitsu and Akira Hirota 1428
Preparation of Epimers of Tea Catechins by Heat Treatment
Ryota Seto, Hironori Nakamura, Fumio Nanjo, and Yukihiko Hara 1434
New Antimicrobial Substances against Streptomyces scabies
from Rosemary(Rosmarinus officinalis L.)
Makoto Takenaka, Toshiro Watanabe, Kazuo Sugahara, Yasuo Harada,Shigeo
Yoshida, and Fumio Sugawara 1440
Cloning and Nucleotide Sequence of the Putative Polyketide
Synthase Genes for Pradimicin Biosynthesis from Actinomadura hibisca
Tohru Dairi, Yoshimitsu Hamano, Yasuhiro Igarashi, Tamotsu Furumai,
and Toshikazu Oki 1445
Changes in Blood Coagulation, Platelet Aggregation, and
Lipid Metabolism in Rats Given Lipids Containing Docosahexaenoic Acid
Norihito Yamada, Yoshiki Kobatake, Sachie Ikegami, Toshichika Takita,
Masahiro Wada, Jun Shimizu, Yusuke Kanke, and Satoshi Innami 1454
Molecular Cloning and Nucleotide Sequence of the Arginase
Gene of Bacillus brevis TT02-8 and Its Expression in Escherichia coli
Kumiko W. Shimotohno, Ikuko Miwa, and Toyoshige Endo- 1459
Acetic Acid Separation from Anaerobically Treated Palm
Oil Mill Effluent by Ion Exchange Resins for the Production of Polyhydroxyalkanoate
by Alcaligenes eutrophus
Mohd. Ali Hassan, Yoshihito Shirai, Haruo Umeki, Hiroshi Yamazumi,
Sha Jin,
Shuichi Yamamoto, Mohd. Ismail Abdul Karim, Kazuhiro Nakanishi, and Kenji
Hashimoto 1465
A Conjugative Linear Plasmid in Streptomyces laurentii
ATCC31255
Chizuru Kinoshita-Iramina, Maki Kitahara, Katsumi Doi, and Seiya Ogata
1469
The Effects of Corn Peptide Ingestion on Facilitating
Alcohol Metabolism in Healthy Men
Magoichi Yamaguchi, Fumi Nishikiori, Michiko Ito, and Yuji Furukawa
1474
Antioxidative Activity of Sulfur-containing Flavor Compounds
in Garlic
Sun Min Kim, Kikue Kubota, and Akio Kobayashi 1482
Purification and Characterization of Ferredoxin-Sulfite
Reductase from Turnip (Brassica rapa) Leaves and Comparison of Properties
with Ferredoxin-Sulfite Reductase from Turnip Roots
Shunji Takahashi, Wai-Cheung Yip, and Goro Tamura 1486
Molecular Characteristics of Water-soluble Polysaccharide
Extracted from Jelly Fig (Ficus awkeotsang Makino) Seeds
Hiroko Suzuno, Shinichi Kinugasa, Hisae Nakahara, and Akiko Kawabata
1491
Subcellular Location of Polyphenol Oxidase in Apples
Masatsune Murata, Mie Tsurutani, Sonoko Hagiwara, and Seiichi Homma
1495
Expression and Characterization of Sucrose Synthase from
Mung Bean Seedlings in Escherichia coli
Tomonori Nakai, Naoto Tonouchi, Takayasu Tsuchida, Hitoshi Mori, Fukumi
Sakai, and Takahisa Hayashi 1500
Inhibition of Collagenases from Mouse Lung Carcinoma Cells
by Green Tea Catechins and Black Tea Theaflavins
Masaki Sazuka, Hirokazu Imazawa, Yutaka Shoji, Takashi Mita, Yukihiko
Hara, and Mamoru Isemura 1504
Purification and Characterization of Cysteine Proteinase
from a Baculovirus Gene
Saori Takahashi, Souko Ushiyama, Takeo Suzuki, Katsuaki Ogawa, and
Kohei Oda 1507
A Novel Acid Phosphatase from Aspergillus niger KU-8 That
Specifically Hydrolyzes C-6 Phosphate Groups of Phosphoryl Oligosaccharides
Kenji To-o, Hiroshi Kamasaka, Kaname Kusaka, Takashi Kuriki, Takashi
Kometani, and Shigetaka Okada 1512
Ajugatakasins A and B, New Diterpenoids from Ajuga decumbens,
and Feeding Stimulative Activity of Related Neoclerodane Analogs toward
the Turnip Sawfly
Takashi Amano, Ritsuo Nishida, and Yasumasa Kuwahara 1518
Isolation and Some Properties of an Iron-oxidizing Bacterium
Thiobacillus ferrooxidans Resistant to Molybdenum Ion
Ng Kim Yong, Mitsuko Oshima, Robert C. Blake, II, and Tsuyoshi Sugio
1523
Purification and Characterization of an Enzyme That Catalyzes
Ring Cleavage of Aspergillic Acid, from Trichoderma koningii ATCC 76666
Atsuo Nishimura, Fumiki Yoshizako, and Mitsuo Chubachi 1527
Lowering Effect of Phenolic Glycosides on the Rise in
Postprandial Glucose in Mice
Hiroshi Takii, Keitaro Matsumoto, Takashi Kometani, Shigetaka Okada,
and Tohru Fushiki 1531
Effect of a Thyroid Hormone Treatment on Brain Protein
Synthesis in Rats
Kazutoshi Hayase, Yaeko Naganuma, Michie Moriyama, Akira Yoshida, and
Hidehiko Yokogoshi 1536
Emulsion-stabilizing Effect of Bacterial Cellulose
Hiroshi Ougiya, Kunihiko Watanabe, Yasushi Morinaga, and Fumihiro Yoshinaga
1541
Facile Synthesis of 2-Hydroxy-6-methylbenzaldehyde, an
Alarm and Sex Pheromone Component of Astigmatid Mites
Satoshi Noguchi, Naoki Mori, Yasumasa Kuwahara, and Masashi Sato 1546
Increases in Hematopoietic Responses Caused by ภ-Glucans
in Mice
Tomoko Tateishi, Naohito Ohno, Yoshiyuki Adachi, and Toshiro Yadomae
1548
Purification and Characterization of a Cysteine Protease
from Corms of Freesia, Freesia reflacta
Makoto Kaneda, Hiroo Yonezawa, and Tetsuya Uchikoba 1554
Beta Ray-induced Scission of DNA in Tritiated Water and
Protection by a Green Tea Percolate and (-)-Epigallocatechin Gallate
Hiroe Yoshioka, Hiromu Kurosaki, Kouichi Yoshinaga, Kieko Saito, and
Hisashi Yoshioka 1560
Synthesis of (})-Hiburipyranone, a Cytotoxic Metabolite
of Marine Sponge Mycale adhaerens
Kanako Uchida, Hidenori Watanabe, and Kenji Mori (>1๕<)1 1564
Purification and Characterization of Novel Whey Glycoprotein
WGP-88 Which Binds to a Monoclonal Antibody to PAS-4 Glycoprotein in the
Bovine Milk Fat Globule Membrane
Sik Hwangbo, Norihiro Azuma, Jun-ichi Kurisaki, and Choemon Kanno 1568
-Note-
Purification and Some Properties of Lignostilbene-ฟ,ภ-dioxygenase Isozyme
IV from Pseudomonas paucimobilis TMY1009
Shigehiro Kamoda, Tamami Terada, and Yoshimasa Saburi 1575
Orientation of Photosynthetic Reaction Center Reconstituted
in Neutral and Charged Liposomes
Masayuki Hara, Takao Ueno, Takaaki Fujii, Qing Yang, Yasuo Asada, and
Jun Miyake, 1577
Transient Expression of Goat Growth Hormone Gene in Poplar
(Populus alba L.) Protoplasts: A Quick Method for Detection of Foreign
Gene Expression in mRNA Level
Jingbo Qiao, Yasuyuki Ishihara, Hiroyuki Kuroda, Fukumi Sakai, Hiroshi
Sakai, and Tohru Komano 1580
Polyamine Content of Ordinary Foodstuffs and Various Fermented
Foods
Akiko Okamoto, Eriko Sugi, Yukimichi Koizumi, Fujiharu Yanagida, and
Shigezo Udaka 1582
High Rate Production in Static Culture of Bacterial Cellulose
from Sucrose by a Newly Isolated Acetobacter Strain
Yukiko Kojima, Akira Seto, Naoto Tonouchi, Takayasu Tsuchida, and Fumihiro
Yoshinaga 1585
Generation of Basidiomycetous Hyphal Cell-aggregates by
Addition of the Arg-Gly-Asp Motif-containing Fragment of High-molecular-weight
Cell-adhesion Protein MFBA Derived from the Basidiomycete Lentinus edodes
Toru Yasuda, Hiroki Ishihara, Hitoshi Amano, and Kazuo Shishido 1587
High Responsiveness to Thyroid Hormone of Adult Rat Primary
Hepatocytes Cultured on EHS-Gel
Hiroaki Oda, Katsura Nozawa, Fuyuko Miyachi, Ai Shimizu, Yukiko Iwasaki,
and Atsushi Kakinuma 1590
Cloning of a Gene Encoding a Putative Xylanase with a
Cellulose-Binding Domain from Humicola grisea
Hiroshi Iikura, Shou Takashima, Akira Nakamura, Haruhiko Masaki, and
Takeshi Uozumi 1593 Vol. 61. No. 9(1997)
DNA Sequence of Bacillus subtilis (natto) NR-1 ม-Glutamyltranspeptidase
Gene, ggt
Yoshihiro Ogawa, Dai Sugiura, Hiroshi Motai, Katsumi Yuasa, and Yasutaka
Tahara 1596
Nitroguanidination of the Amino Groups of Ribonuclease
T(>21<)2 with N-Methyl-N'-nitro- N-nitrosoguanidine
Kenji Takahashi 1601
Effects of Benadrostin, a Poly (ADP-Ribose) Polymerase
Inhibitor, on Apoptosis of HL-60 Induced by 5-Azacytidine
Hiroyasu Tobe, Mitsune Yamaguchi, Tomoji Kocha, and Takaaki Aoyagi
1603
Identification of Cyanidin 3-O-(3'',6''-O-Dimalonyl-ภ-glucopyranoside)
as a Flower Pigment of Chrysanthemum (Dendranthema grandiflorum)
Masayoshi Nakayama, Masaji Koshioka, Michio Shibata, Syuntaro Hiradate,
Hajime Sugie, and Masa-atsu Yamaguchi 1607
-Rapid Paper-
Protease Catalyzed Reaction of Polyamine Incorporation into Protein
Shunichi Suzuki, Katsunori Kobayashi, Shigeru Yamanaka, and Kenzo Yokozeki
1609
-Short Communication-
A Ribosome-Inactivating Protein from Amaranthus viridis
Seok-Yoon Kwon, Chung Sun An, Jang Ryol Liu, and Kyung-Hee Paek 1613
Synthesis of 2-O-Fucosyl Sulfatide, a Blocker of L- and
P-Selectin
Hideharu Ishida, Satoshi Sago, Takao Ikami, Makoto Kiso, and Akira Hasegawa
1615
Distinguishable Action between Acid-stable and Neutral
ฟ-Amylases from Shochu Koji (Aspergillus kawachii )
Toshihiko Suganuma, Naoyuki Noda, Hiroyuki Honbo, and Kanefumi Kitahara
1617
-1-
Review
Biocatalysis in Organic Synthesis: The Use of Nitrile- and Amide-hydrolyzing
Microorganisms
Takeshi Sugai,๕ Takahiro Yamazaki, Masahiro Yokoyama, and Hiromichi Ohta๕
Department of Chemistry, Keio University, 3-14-1 Hiyoshi, Yokohama 223, Japan
@This review covers recent examples of synthetic transformation of
nitriles and amides by microbial enzyme systems. A variety of substrates
and products involving enantiomerically enriched forms of chiral substances
are referred to. The stereochemical course of enzyme-catalyzed hydrolysis
is briefly commented on. Special emphasis is placed upon the range of functional
groups that are acceptable by enzymes and/or survive under the transformation,
as well as the advantages as a synthetic tool for conversion under mild
conditions.
Key words: nitrile; amide; hydrolysis; microorganisms; organic synthesis
-2-
Two Oxazolyl Compounds and a Monosubstituted ฟ-Pyrone
as Free-radical Scavengers Isolated from a Fungus
Yasujiro Morimitsu ๕,๕๕ and Akira Hirota
Laboratory of Applied Microbiology, School of Food and Nutritional Sciences, University of Shizuoka, 52-1 Yada, Shizuoka 422, Japan
Received July 22, 1996
@In the course of our screening for free radical scavengers, (1'E )-erythro-4-(3',4'-dihydroxypentenyl)oxazole
(1) (1'E,4'S )-4-(3'-oxo-4'-hydroxypentenyl)oxazole (2) and 6-pentyl-ฟ-pyrone
(3) were isolated from an unidentified fungal metabolite. These compounds,
especially novel oxazolyl compound 2, inhibited the bactericidal effect
of the Fenton reagent toward Bacillus subtilis. They and their acetylated
compounds (diAc-1 and Ac-2) also showed inhibitory activity against linoleate
autoxidation. Furthermore, 1-3 inhibited oxidative enzymes (soybean lipoxygenase
and mushroom tyrosinase). To investigate the radical scavenging mechanism
of 3, two oxidized products (4 and 5) were isolated from the reaction mixture
of 3 and the Fenton reagent. Compounds 4 and 5 seemed to be derived from
3 by scavenging the hydroxyl radical.
Key words: radical scavenger; Fenton reaction; bactericidal action; fungal
metabolite; oxazolyl compound
-3-
Preparation of Epimers of Tea Catechins by Heat Treatment
Ryota Seto, Hironori Nakamura, Fumio Nanjo, and Yukihiko Hara
Food Research Laboratories, Mitsui Norin Co., Ltd., 223-1 Miyabara, Fujieda-shi, Shizuoka 426-01, Japan
Received September 17, 1996
@Products from tea catechins ((+)-catechin, (-)-epicatechin, (-)-epigallocatechin,
(-)-epicatechin gallate, and (-)-epigallocatechin gallate) after heat treatment
were individually isolated by preparative HPLC and subsequent crystallization.
FAB-MS, elemental, 1H- and 13C-NMR, and optical rotation analyses indicated
these products to be the C-2 epimers of the original catechins. The effects
of temperature, time, pH, and concentration on this conversion of tea catechins
to their epimers were examined. It was determined that the epimerization
reaction should be conducted with 1% each of the tea catechin solution
(pH 5) at 120C for 30 min.
Key words: tea catechin; epimerization; tea catechin epimer; heat treatment
-4-
New Antimicrobial Substances against Streptomyces scabies
from Rosemary (Rosmarinus officinalis L.)
Makoto Takenaka,๕ Toshiro Watanabe,* Kazuo Sugahara,๕๕ Yasuo Harada,๕๕๕ Shigeo Yoshida,** and Fumio Sugawara***
National Institute of Agro-Environmental Sciences, Tsukuba, Ibaraki
305, Japan
* Fukuoka Agricultural Research Center, Chikushino, Fukuoka 818, Japan
** The Institute of Physical and Chemical Research (RIKEN ), Wako, Saitama
351-01, Japan
*** Department of Applied Bioscience, Science University of Tokyo, Noda,
Chiba 278, Japan
Received September 27, 1996
@New antimicrobial substances against Streptomyces scabies, rosmic acid
and rosmanol-related compounds, were isolated from leaves of rosemary (Rosmarinus
officinalis L.). The active compounds were separated in pure form by a
combination of gel permeation chromatography and reverse-phase HPLC. Their
structures were determined by two-dimensional NMR experiments and HR-MS
analyses.
Key words: Streptomyces scabies; Rosmarinus officinalis L; rosmic acid;
rosmanol; antimicrobial substance
-5-
Cloning and Nucleotide Sequence of the Putative Polyketide
Synthase Genes for Pradimicin Biosynthesis from Actinomadura hibisca ๕
Tohru Dairi,๕๕ Yoshimitsu Hamano, Yasuhiro Igarashi, Tamotsu Furumai, and Toshikazu Oki
Toyama Prefectural University, Biotechnology Research Center, 5180 Kurokawa, Kosugi, Toyama 939-03, Japan
Received November 13, 1996
@We cloned the putative polyketide synthase genes ( pms genes) for pradimicin
A biosynthesis from Actinomadura hibisca using an oligonucleotide probe
designed on the basis of conserved amino acid sequences of other polyketide
synthases (PKSs). By DNA sequencing of an 8.2-kb SacI fragment that hybridized
with the oligonucleotide probe, 11 open reading frames (ORFs) were found.
All of the ORFs except for ORF10 were predicted to be translated in the
same direction. Each of the deduced ORFs has significant sequence similarity
to the protein responsible for polyketide biosynthesis or spore pigmentation.
In particular, ORF1, ORF2, and ORF3 were 50-70% identical with genes coding
for PKSs for actinorhodin biosynthesis. Specific DNA regions similar in
sequence to pms genes were found with genomic Southern hybridization in
all of the pradimicin producers examined, but were not found in pradimicin
nonproducers, suggesting that the genes cloned in this study encode polyketide
synthase for pradimicin biosynthesis.
Key words: Actinomadura hibisca; pradimicin; polyketide synthase
-6-
Changes in Blood Coagulation, Platelet Aggregation, and
Lipid Metabolism in Rats Given Lipids Containing Docosahexaenoic Acid
Norihito Yamada,๕ Yoshiki Kobatake,* Sachie Ikegami,** Toshichika Takita, Masahiro Wada, Jun Shimizu, Yusuke Kanke, and Satoshi Innami
Department of Nutrition, Faculty of Agriculture, Tokyo University of
Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156, Japan
* Department of Nutrition, Chiba Prefectural Hygiene Junior College, 2-10-1
Wakaba, Mihama-ku, Chiba 261, Japan
** Division of Food Science, The National Institute of Health and Nutrition,
Tokyo 162, Japan
Received December 16, 1996
@In order to identify an adequate intake level of docosahexaenoic acid
(DHA), changes in various parameters related to health benefits were studied
in rats fed on diets containing 10% test lipids at different n-3(DHA)/n-6
ratios for two weeks.
@An evaluation of the critical level of the dietary n-3/n-6 ratio which
had a significant effect on the parameters of several tissues indicated
that the response to the dietary ratios differed according to the parameter,
the variation in ratio ranging approximately from 0.20 to 1.77 with either
a positive or negative effect on the health benefit. These results suggest
that a suitable intake level of DHA would be within this range.
@In view of safety, however, the critical level for the dietary n-3/n-6
ratio may be around 0.56, as shown by a detailed analysis on the lower
limit level of the harmful parameters. We thus propose that the dietary
intake of DHA should not be more than 0.56 in terms of the n-3/n-6 ratio.
Key words: docosahexaenoic acid; platelet aggregation; blood coagulation;
plasma and liver lipids; lipid peroxides
-7-
Molecular Cloning and Nucleotide Sequence of the Arginase
Gene of Bacillus brevis TT02-8 and Its Expression in Escherichia coli ๕
Kumiko W. Shimotohno,๕๕ Ikuko Miwa, and Toyoshige Endo-
Kyoritsu College of Pharmacy, 1-5-30 Shiba-koen, Minato-ku, Tokyo 105, Japan
Received December 17, 1996
@The gene from Bacillus brevis TT02-8 encoding arginase was cloned into
Escherichia coli, and its nucleotide sequence was identified. The nucleotide
sequence contained an open reading frame that encoded a polypeptide of
298 amino acid residues with a predicted molecular weight of 31,891, which
was consistent with that previously calculated for arginase purified from
this bacterium. Comparison of the deduced amino acid sequence of the B.
brevis TT02-8 arginase with that of the prokaryotic and eukaryotic arginases
of Bacillus caldovelox, Bacillus subtilis, Agrobacterium Ti plasmid C58,
Saccharomyces cerevisiae, Coccidioides immitis, Xenopus laevis, Rana catesbeiana,
rat liver, and human liver, showed 33-66% of the sequences to be similar;
there were several highly conserved regions. Arginase activity was detected
in Escherichia coli cells transformed with an expression plasmid of the
cloned arginase gene.
Key words: arginase gene of Bacillus brevis; gene cloning; sequence similarity
-8-
Acetic Acid Separation from Anaerobically Treated Palm
Oil Mill Effluent by Ion Exchange Resins for the Production of Polyhydroxyalkanoate
by Alcaligenes eutrophus
Mohd. Ali Hassan,1 Yoshihito Shirai,2,๕ Haruo Umeki,2 Hiroshi Yamazumi,2 Sha Jin,3 Shuichi Yamamoto,4 Mohd. Ismail Abdul Karim,1 Kazuhiro Nakanishi,5 and Kenji Hashimoto6
1 Department of Biotechnology, University Putra Malaysia, 43400 UPM
Serdang, Selangor, Malaysia
2 Department of Biochemical Engineering and Science, Kyushu Institute of
Technology, Iizuka, Fukuoka 820, Japan
3 National Key Laboratory of Bioreactors, East China University of Science
and Technology, Shanghai 200237, China
4 Department of Chemical Engineering, Yamaguchi University, Tokiwadai,
Ube 755, Japan
5 Department of Biotechnology, Okayama University, Tsushima-naka, Okayama
700, Japan
6 Department of Chemical Engineering, Kyoto University, Sakyo-ku, Kyoto
606, Japan
Received December 18, 1996
@Separation of acetic acid from palm oil mill effluent (POME) to increase
its concentration by an anion exchange resin was examined as a preliminary
study for its recovery from POME that had been anaerobically treated by
sludge from a palm oil mill. This paper concerns the acetic acid thus separated
for producing bacterial polyhydroxyalkanoate (PHA) by Alcaligenes eutrophus.
It was found that sludge particles in POME strongly inhibited the adsorption
of acetic acid on the anion exchange resin. Removing the sludge particles
from the POME facilitated the separation of acetic acid from the POME efficiently.
The concentrated acetic acid thus obtained from anaerobically treated POME
could be used as a substrate in the fed-batch production of polyhydroxyalkanoate
by Alcaligenes eutrophus.
Key words: acetic acid; ion-exchange separation; palm oil mill effluent;
polyhydroxyalkanoates; fed-batch culture
-9-
A Conjugative Linear Plasmid in Streptomyces laurentii
ATCC31255
Chizuru Kinoshita-Iramina, Maki Kitahara, Katsumi Doi, and Seiya Ogata๕
Microbial Genetics Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University, Hakozaki, Higashi-ku, Fukuoka 812-81, Japan
Received December 24, 1996
@Plasmid pSLL of Streptomyces laurentii ATCC31255 (wild-type strain P0)
is a 93-kilobase linear DNA plasmid that carries a protein bound to each
5' end of the DNA. It was self-transmitted to the pSLL-cured strain by
conjugation in solid culture. The pSLL-cured strain carried a circular
plasmid, pSLS, and showed a marked decrease in spore formation and thiostrepton
productivity, owing to the pSLS. However, by retransmission of pSLL, these
things reverted to levels seen in strain P0. Thus, plasmid pSLL suppressed
the injurious effects of pSLS on the host mycelia.
Key words: Streptomyces; linear plasmid; conjugative plasmid
-10-
The Effects of Corn Peptide Ingestion on Facilitating
Alcohol Metabolism in Healthy Men
Magoichi Yamaguchi, Fumi Nishikiori, Michiko Ito,* and Yuji Furukawa*
Research Institute, Nihon Shokuhin Kako Co., Ltd., Tajima, Fuji 417,
Japan
* Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku
University, Aoba-ku, Sendai 981, Japan
Received January 9, 1997
@We prepared corn peptide (CP), a vegetable oligopeptide and tried to
discover the effects of its ingestion on facilitating alcohol metabolism
in healthy adult men. Ten healthy male volunteers ingested 5 g of CP, wheat
peptide (WP), pea peptide (PP), alanine, or leucine 30 min before alcohol
intake at a dose of 0.5 g/kg, and blood ethanol and plasma amino acid concentrations
were measured during a 2-h observation period after alcohol intake. In
subjects who ingested CP, the blood ethanol level was lower than that in
the WP, alanine and leucine ingestion groups, but did not decrease as compared
to the control when they ingested PP. Similarly there was a difference
in the blood ethanol level between alanine and leucine ingestion groups,
and leucine ingestion was more effective than alanine against the reduction
of the increase in blood ethanol level. On the other hand, there was no
significant difference in the plasma concentrations of individual amino
acids except alanine, leucine, or lysine after alcohol intake among experimental
groups as compared to the control. CP ingestion significantly elevated
plasma alanine and leucine rather than other groups during a 2-h observation
period. These results suggested that CP may have the effect on the reduction
of increase in blood ethanol level after alcohol intake by the marked elevation
of plasma alanine and leucine, especially leucine, but neither by the delay
of ethanol release from the stomach nor malabsorption of ethanol in the
gastrointestinal tracts.
Key words: corn peptide; blood ethanol; plasma amino acid; alanine; leucine
-11-
Antioxidative Activity of Sulfur-containing Flavor Compounds
in Garlic
Sun Min Kim, Kikue Kubota,* and Akio Kobayashi*
Department of Food and Biotechnology, Dongshin University, 252 Daehodong,
Naju, Chonnam 520-714, Republic of Korea
* Laboratory of Food Chemistry,
Department of Nutrition and Food Science, Ochanomizu University, 2-1-1
Otsuka, Bunkyo-ku, Tokyo 112, Japan
Received January 9, 1997
@The antioxidative activity of garlic homogenates blended with distilled
water and different amounts of soybean oil was examined. The antioxidative
activity of garlic aroma concentrate and that of the oil layer from the
garlic homogenate was strong. The stability of the three sulfur compounds
in garlic, allicin, diallyl disulfide, and diallyl trisulfide, was analyzed
by HPLC, diallyl disulfide being the most stable. It was found that the
effectiveness and strength of diallyl disulfide as an antioxidant could
be increased by additing ฟ-tocopherol and L-ascorbyl palmitate in a lard
system.
Key words: antioxidative activity of garlic; diallyl disulfide; antioxidative
synergism; garlic flavor; allicin
-12-
Purification and Characterization of Ferredoxin-Sulfite
Reductase from Turnip (Brassica rapa) Leaves and Comparison of Properties
with Ferredoxin-Sulfite Reductase from Turnip Roots
Shunji Takahashi, Wai-Cheung Yip, and Goro Tamura๕
Department of Bioresources Chemistry, Faculty of Horticulture, Chiba University, Matsudo 648, Chiba 271, Japan
Received January 20, 1997
@Ferredoxin-sulfite reductase (Fd-SiR) [hydrogen-sulfide: ferredoxin oxidoreductase,
EC 1.8.7.1] from turnip leaves (SiR-L) has been purified to homogeneity
and its enzymatic properties compared with that from turnip roots (SiR-R).
Each enzyme had a molecular mass of 64.5}0.5 kDa by SDS-PAGE and an isoelectric
point of 5.15}0.05. Although each had a pH optimum around 7.8 with the
same effects of inhibitors, SiR-L had higher heat stability at 60C than
SiR-R. Moreover, SiR-R had a lower Km and a higher specificity constant
(k(/)cat /Km) for turnip leaf ferredoxin than SiR-L. The N-terminal amino
acid sequence of SiR-L was different from that of SiR-R. The results of
amino acid analysis and peptide mapping suggested that SiR-L and SiR-R
have different primary structures.
Key words: ferredoxin-dependent enzyme; non-photosynthetic tissue; siroheme;
sulfate assimilation; sulfite reductase
-13-
Molecular Characteristics of Water-soluble Polysaccharide
Extracted from Jelly Fig (Ficus awkeotsang Makino) Seeds
Hiroko Suzuno, Shinichi Kinugasa,* Hisae Nakahara,* and Akiko Kawabata**
Department of Nutrition, Junior College of Tokyo University of Agriculture,
1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156, Japan
* National Institute of Materials and Chemical Research, 1-1 Higashi, Tsukuba,
Ibaraki 305, Japan
** Department of Nutrition, Faculty of Agriculture, Tokyo University of
Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156, Japan
Received January 31, 1997
@The molecular characteristics of polysaccharide obtained by extracting
with water from jelly fig (Ficus awkeotsang Makino) seeds were elucidated
by measuring the weight-average molecular weight (Mw) and radius of gyration
(RG), indicating the molecular expanse, by the light-scattering method.
@The Mw and RG values of the water-soluble jelly fig polysaccharide were
84-130~104 and 240-450 mm, respectively, these values being larger than
those of commercial LM pectin and HM pectin. In addition, the Mw and RG
values of the water-soluble jelly fig polysaccharide initially increased
after being extracted, indicating the highest values 5 hours after the
extraction, and thereafter decreased. These changes in the time-course
of the molecule reflected well the changes with time in the mechanical
characteristics and network structure of the water-soluble jelly fig polysaccharide
gel. Exponent ฟ in the expression RGๅMwฟ was found to be 0.37. From
these results, the conformation of the water-soluble jelly fig polysaccharide
molecule after association by the contained inorganic elements proved to
be of globular form rather than a random coil shape as a result of contraction
of the molecule.
Key words: jelly fig (Ficus awkeotsang Makino); light-scattering measurement;
molecular weight; radius of gyration; galacturonic acid
-14-
Subcellular Location of Polyphenol Oxidase in Apples
Masatsune Murata,๕ Mie Tsurutani, Sonoko Hagiwara, and Seiichi Homma
Department of Nutrition and Food Science, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo 112, Japan
Received February 3, 1997
@The location of polyphenol oxidase (PPO) in cells of apple fruit was
examined by immunohistochemistry and subcellular fractionation. In mature
apple fruits, where vacuoles occupy most of the cells, PPO was detected
immunochemically near the cell walls with use of anti-apple PPO antibodies.
In cells of immature fruits and tissue culture, PPO was detected in organelles
other than the vacuoles, probably in plastids. The plastid fraction was
purified by density gradient ultracentrifugation, and the activities of
PPO and marker enzymes of plastids were the highest in the plastids. Most
apple PPO was in plastids, as are other plant PPOs, and some of the protein
was solubilized and proteolyzed during ripening and storage.
Key words: polyphenol oxidase; apple (Malus pumila); plastid; subcellular
location; enzymatic browning
-15-
Expression and Characterization of Sucrose Synthase from
Mung Bean Seedlings in Escherichia coli
Tomonori Nakai, Naoto Tonouchi,* Takayasu Tsuchida,* Hitoshi Mori,** Fukumi Sakai, and Takahisa Hayashi๕
Wood Research Institute, Kyoto University, Gokasho, Uji, Kyoto 611,
Japan
* Bio-Polymer Research Co., Ltd., KSP R & D Business-park Bldg. B-1015,
2-1, Sakato 3-chome, Takatsu-ku, Kawasaki 213, Japan
** Faculty of Agriculture, Nagoya University, Chikusa, Nagoya 464-01, Japan
Received February 12, 1997
@The cDNA fragment coding for mung bean (Vigna radiata Wilczek) sucrose
synthase was introduced into the expression vector pET-20b resulting in
the construction of plasmid pEB-01. After transformation of Escherichia
coli strain BL21(DE3) cells by pEB-01 and induction with isopropyl thio-ภ-galactoside,
high level expression of the recombinant enzyme was obtained. The enzyme
had a tetrameric form that conserved the activity of sucrose synthase.
Although the Km and V(/)max of the recombinant enzyme acting on either
UDP-glucose or fructose were very close to those of the native enzyme isolated
from mung bean seedlings, the Km for sucrose was higher by a factor of
10 for the recombinant enzyme. This suggests that the recombinant sucrose
synthase has a tendency to synthesize sucrose, although the native enzyme
catalyzes a freely reversible reaction.
Key words: mung bean; recombinant; sucrose synthase; kinetics
-16-
Inhibition of Collagenases from Mouse Lung Carcinoma Cells
by Green Tea Catechins and Black Tea Theaflavins
Masaki Sazuka, Hirokazu Imazawa, Yutaka Shoji, Takashi Mita, Yukihiko Hara,* and Mamoru Isemura
School of Food and Nutritional Sciences, University of Shizuoka, Yada,
Shizuoka 422, Japan
* Food Research Laboratories, Mitsui Norin Co., Ltd., 2-1-17 Fujieda, Shizuoka
426, Japan
Received February 14, 1997
@Theaflavin and theaflavin digallate, which are components of black tea
were examined by in vitro invasion assay with mouse Lewis lung carcinoma
LL2-Lu3 cells, which are highly metastatic. The compounds inhibited invasion
by the tumor cells. Gelatin zymography showed that the cells secreted matrix
metalloproteinases (MMPs), probably including MMP-2 and MMP-9, which may
be involved in tumor cell invasion and metastasis. Theaflavin and theaflavin
digallate also inhibited MMPs from the culture medium of these tumor cells,
as did (-)-epigallocatechin gallate. These results suggest that theaflavin,
theaflavin digallate, and (-)-epigallocatechin gallate inhibit tumor cell
invasion by inhibiting type IV collagenases of the LL2-Lu3 cells.
Key words: (-)-epigallocatechin gallate; theaflavin; theaflavin digallate;
matrix metalloproteinase; invasion
-17-
Purification and Characterization of Cysteine Proteinase
from a Baculovirus Gene
Saori Takahashi,๕ Souko Ushiyama, Takeo Suzuki,* Katsuaki Ogawa,* and Kohei Oda
Department of Applied Biology, Faculty of Textile Science, Kyoto Institute
of Technology, Matsugasaki, Sakyo-ku, Kyoto 606, Japan
* Katakura Industries Co., Ltd., Tyuou, Matumoto, Nagano 390, Japan
Received February 14, 1997
@To analyze the degradation of product proteins at the late stage of virus
infection in the baculovirus expression system, a cysteine proteinase was
purified from hemolymph of Bombyx mori infected with wild-type B. mori
nuclear polyhedorosis virus (BmNPV). The purified cysteine proteinase preparation
had two protein bands (major 35-kDa active protein and 28-kDa inactive
protein) on SDS-PAGE. Based on the N-terminal amino acid sequences of them,
it was found that both proteins originated in the cysteine proteinase gene
of BmNPV. The purified cysteine proteinase had an optimum pH at 4.0, and
also had activities at neutral pHs. When recombinant luciferase was used
as a natural substrate, it was degraded rapidly by the cysteine proteinase
at the physiological pH of hemolymph. These results suggest that the cysteine
proteinase from a BmNPV gene participates in the degradation of foreign
protein expressed by the baculovirus system.
Key words: baculovirus; cysteine proteinase; purification; characterization
-18-
A Novel Acid Phosphatase from Aspergillus niger KU-8 That
Specifically Hydrolyzes C-6 Phosphate Groups of Phosphoryl Oligosaccharides
Kenji To-o, Hiroshi Kamasaka, Kaname Kusaka, Takashi Kuriki, Takashi Kometani, and Shigetaka Okada
Biochemical Research Laboratory, Ezaki Glico Co., Ltd., 4-6-5 Utajima, Nishiyodogawa-ku, Osaka 555, Japan
Received March 5, 1997
@We had analyzed the detailed structures of the phosphoryl oligosaccharide-1
(PO-1) fraction that was the main component of phosphoryl oligosaccharides
(POs) prepared from a potato starch hydrolysate. PO-1 fraction was made
up of 3-phosphoryl oligosaccharides and 6-phosphoryl oligosaccharides.
Aspergillus niger strain KU-8 produced two types of intracellular acid
phosphatase (EC 3.1.3.2, ACPase); ACPase I and II. ACPase II preferentially
dephosphorylated 6-phosphoryl oligosaccharides rather than 3-phosphoryl
oligosaccharides. The molecular weight of the enzyme was estimated as 66
kDa by SDS-polyacrylamide gel electrophoresis and about 260 kDa by gel
filtration, implying the active form to be a tetramer. The optimum pH and
temperature of the enzyme were 2.0-2.5 and 60C, respectively. ACPase
II was stable below 50C for 30 min and pH 2.0-10.0 for 60 min. In spite
of the strict specificity toward 6-phosphoryl oligosaccharides in the PO-1
fraction, ACPase II was able to hydrolyze Fru-1,6-di-P, ATP, pyrophosphate,
and polyphosphate as well as pNPP and Glc-6-P, a broad substrate specificity.
Key words: potato starch; phosphoryl oligosaccharides; Aspergillus niger;
acid phosphatase; substrate specificity
-19-
Ajugatakasins A and B, New Diterpenoids from Ajuga decumbens,
and Feeding Stimulative Activity of Related Neoclerodane Analogs toward
the Turnip Sawfly
Takashi Amano,๕ Ritsuo Nishida,๕๕ and Yasumasa Kuwahara
Pesticide Research Institute, Faculty of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan
Received March 6, 1997
@Adults of the turnip sawfly, Athalia rosae ruficornis, are strongly attracted
to the leaves of Ajuga decumbens (Labiatae). Specific feeding stimulants
of the sawfly were examined in the leaf surface extracts of A. decumbens.
Among seven neoclerodane diterpenoids isolated from the extracts, two new
analogs, ajugatakasins A and B, were characterized as 6ฟ,18-diacetoxy-1ภ,12-ditigloyloxy-4ฟ,17-epoxyneoclerod-13-en-15,16-olide
and its 1ภ,12-di-(2-methylbutanoyl)-oxy analog, respectively. Among these
analogs, clerodendrin D, a very minor constituent which possesses a tetrahydrofurofuran
moiety, was identified as the feeding stimulant for A. rosae ruficornis,
while the other analogs with an ฟ,ภ-unsaturated ม-lactone moiety were
found to be completely inactive in the sawfly feeding test.
Key words: ajugatakasin; neoclerodane; turnip sawfly; Athalia rosae ruficornis
; Ajuga decumbens
-20-
Isolation and Some Properties of an Iron-oxidizing Bacterium
Thiobacillus ferrooxidans Resistant to Molybdenum Ion
Ng Kim Yong, Mitsuko Oshima,* Robert C. Blake, II,** and Tsuyoshi Sugio๕
Department of Biological Function and Genetic Resources Science, Faculty
of Agriculture, Okayama University, 1-1-1 Tsushima Naka, Okayama 700, Japan
* Department of Analytical Chemistry, Faculty of Science, Okayama University,
3-1-1 Tsushima Naka, Okayama 700, Japan
** Xavier University, New Orleans, Louisiana, U.S.A.
Received March 13, 1997
@Among seventy five strains of iron-oxidizing bacteria obtained from natural
environments, only one strain, Thiobacillus ferrooxidans Funis 2-1, grew
on Fe2+-medium with 1.25 mM of sodium molybdate (Mo6+). In contrast, T.
ferrooxidans AP19-3, the representative of Mo sensitive strains, could
not grow on Fe2+-medium with 1.0 mM of sodium molybdate. By comparing the
levels of inhibition of iron oxidase and cytochrome c oxidase by Mo6+ or
Mo5+, it was found that Mo5+ but not Mo6+ is an actual inhibitor for the
iron oxidation enzyme system, especially for cytochrome c oxidase. Cytochrome
c oxidase of Funis 2-1 was more resistant to Mo5+ than AP19-3. Mo5+, compared
to Mo6+, strongly binds to both cells and the plasma membrane of T. ferrooxidans.
Funis 2-1 cells showed a lower binding activity to Mo6+ or Mo5+ compared
to AP19-3. Cytochrome c oxidase of T. ferrooxidans has been known to catalyze
the oxidation of not only reduced mammalian cytochrome c but also Mo5+.
Mo5+-oxidizing activities measured with intact cells and a purified cytochrome
c oxidase from Funis 2-1 cells were higher than those of AP19-3, suggesting
that Funis 2-1 cells can oxidize toxic Mo5+ more rapidly to harmless Mo6+
than AP19-3 does. Since Mo6+ is known to be chemically reduced by Fe2+
to give Mo5+ and Fe3+, the growth inhibition by sodium molybdate (Mo6+)
observed in T. ferrooxidans is explained as follows: Mo6+ added to Fe2+-medium
is chemically reduced by Fe2+, and Mo5+ thus produced binds to the plasma
membrane and inhibits iron oxidase, as a result, growth of the bacterium
is stopped.
Key words: iron-oxidizing bacterium; Thiobacillus ferrooxidans ; molybdenum;
resistance
-21-
Purification and Characterization of an Enzyme That Catalyzes
Ring Cleavage of Aspergillic Acid, from Trichoderma koningii ATCC 76666
Atsuo Nishimura,๕ Fumiki Yoshizako, and Mitsuo Chubachi
Research Institute for Advanced Science and Technology, Osaka Prefecture University, 1-2 Gakuen-cho, Sakai, Osaka 593, Japan
Received March 17, 1997
@The aspergillic acid degrading enzyme (ADE) that catalyzes the cleavage
of the pyrazine ring in aspergillic acid (AA, 1-hydroxy-3-isobutyl-6-sec-butyl-2-pyrazinone)
was purified to electrophoretic homogeneity from extracts of Trichoderma
koningii ATCC 76666. ADE was a homodimeric protein with a molecular mass
of 112 kDa, contained 1 mol of FAD per mol of subunit, and required NAD(P)H
and molecular oxygen for its activity. ADE had an isoelectric point of
around 5.3, and an optimum pH of 7.0-8.0. p-Chloromercuribenzoate and HgCl2
completely inhibited ADE activity, while metal chelating reagents, ฟ,ฟ
'-dipyridyl and o-phenanthroline, were not inhibitors. The substrate specificity
among AA-related compounds was that hydroxyaspergillic acid was a poor
substrate (16% of the activity for AA) and deoxyaspergillic acid did not
serve as a substrate.
Key words: Aspergillic acid degradation; ring cleavage enzyme; Trichoderma
koningii
-22-
Lowering Effect of Phenolic Glycosides on the Rise in
Postprandial Glucose in Mice
Hiroshi Takii,๕ Keitaro Matsumoto,* Takashi Kometani, Shigetaka Okada, and Tohru Fushiki*
Biochemical Research Laboratory, Ezaki Glico Co., Ltd., 4-6-5 Utajima,
Nishiyodogawa-ku, Osaka 555, Japan
* Laboratory of Nutritional Chemistry, Department of Food Science and Technology,
Faculty of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku,
Kyoto 606, Japan
Received March 24, 1997
@Glycosides were screened for their lowering effect on the postprandial
blood glucose rise in vivo. The effect of phlorizin and other phenolic
glycosides on the postprandial blood glucose response to glucose ingestion
was evaluated in Std ddY mice. When phlorizin was simultaneously added,
the peak blood glucose level was significantly decreased by 51% ( p<0.01)
compared to vehicles following glucose ingestion by mice, while the blood
insulin responses were generally similar. Screening experiments were conducted
with different classes of phenolic glycosides added to a glucose solution.
Reductions of 40-52% ( p<0.05) were observed in vehicles containing
arbutin, 4-hydroxyphenyl-ฟ-D-glucopyranoside (hydroquinone-ฟ-glucoside)
or glycyrrhizin, and of only 15-31% (not significant) in vehicles containing
neohesperidin dihydrochalcone, glycyrrhetinic acid monoglucuronide, or
3,4-dimethoxyphenyl-ภ-D-glucopyranoside. No lowering effect was observed
in vehicles containing salicin. Since glycyrrhizin, arbutin, and hydroquinone-ฟ-glucoside
blunted to varying degrees the postprandial blood glucose rise following
glucose ingestion, they may be useful adjuvants for the treatment of diabetic
subjects.
Key words: phlorizin; glycosides; mice; hypoglycemic effect; postprandial
blood glucose
-23-
Effect of a Thyroid Hormone Treatment on Brain Protein
Synthesis in Rats
Kazutoshi Hayase,๕ Yaeko Naganuma, Michie Moriyama,* Akira Yoshida,** and Hidehiko Yokogoshi***
Department of Home Economics, Aichi University of Education, Kariya,
Aichi 448, Japan
* Department of Home Economics, Aichi Gakusen College, Okazaki 444, Japan
** Department of Food and Nutritional Sciences, Nagoya Bunri Junior College,
Nagoya 451, Japan
*** Laboratory of Nutritional Biochemistry, School of Food and Nutritional
Sciences, The University of Shizuoka, Shizuoka 422, Japan
Received March 24, 1997
@The effect of the thyroid hormone on the rate of brain protein synthesis
in rats was studied. Experiments were conducted on three groups of rats
given 6-propyl-2-thiouracil (PTU, a thyroid inhibitor) without a triiodothyronine
(T3) treatment, those treated with PTU+T3, and those treated with neither
PTU nor T3 (control). The fractional rates of protein synthesis in the
brain, liver, and kidney of rats given PTU+T3 were significantly greater
than those in rats given PTU alone. In the brain and kidney, the RNA activity
[g of protein synthesized/(g of RNAEd)] were significantly correlated
with the fractional rates of protein synthesis. In the liver and kidney,
the RNA concentration (mg of RNA/g of protein) was related to the fractional
rate of protein synthesis. These results suggest that the thyroid hormone
treatment would be likely to increase the rate of protein synthesis in
the brain of rats, and that the RNA activity is, at least partly, related
to the fractional rate of brain protein synthesis.
Key words: triiodothyronine; protein synthesis; brain; RNA activity; thyroid
hormone
-24-
Emulsion-stabilizing Effect of Bacterial Cellulose
Hiroshi Ougiya, Kunihiko Watanabe, Yasushi Morinaga, and Fumihiro Yoshinaga
Bio-Polymer Research Co., Ltd., KSP R & D Business-park Bldg. B-1015, 3-2-1 Sakato, Takatsu-ku, Kawasaki-shi, Kanagawa 213, Japan
Received March 24, 1997
@The bacterial cellulose produced in an agitated culture (Ag-BC) showed
the highest emulsion-stabilizing effect among the examined cellulosic materials.
It was clarified that a mechanical barrier and a scaffolding structure
composed of fine fibrils of bacterial cellulose interrupted the coalescence
of oil droplets to stabilize the emulsion without reducing the interfacial
tension as occurred with sorbitan monolaurate. Since Ag-BC consists of
thinner fibrils and smaller flocs than any other cellulosic material, Ag-BC
would cover a larger surface area of the oil droplet as a mechanical barrier.
@The emulsion containing Ag-BC was stable against the addition of salt,
and changes in pH and temperature in comparison with xanthan gum and sorbitan
monolaurate. This stability would have been due to the stability of the
mechanical barrier and a scaffolding structure composed of stable crystalline
cellulose. In contrast, instability in the conformation of xanthan gum
and a reduction in the interfacial tension of the surfactant would lead
to instability of the emulsion.
Key words: bacterial cellulose; cellulose fiber; emulsion-stabilizing effect;
mechanical barrier; scaffolding structure
-25-
Facile Synthesis of 2-Hydroxy-6-methylbenzaldehyde, an
Alarm and Sex Pheromone Component of Astigmatid Mites๕
Satoshi Noguchi,๕๕ Naoki Mori, Yasumasa Kuwahara,๕๕๕ and Masashi Sato*
Pesticide Research Institute, Faculty of Agriculture, Kyoto University,
Sakyo-ku, Kyoto 606-01, Japan
* Department of Bioresource Science, Faculty of Agriculture, Kagawa University,
Miki-cho, Kagawa 761-07, Japan
Received March 24, 1997
@2-Hydroxy-6-methylbenzaldehyde (5) is a component of astigmatid mites,
functioning as the alarm and sex pheromones, and the establishment of its
convenient synthesis is prerequisite and of vital importance to develop
applications of these pheromones for practical use.
@The target compound (5) was effectively synthesized from m-cresol (1)
by the following four steps in a 44% overall yield: protection of the hydroxyl
group by a tetrahydropyranyl group, blocking the active site (6-position
of 1) by a trimethylsilyl group via lithiation and subsequent silylation,
formylation via lithiation and successive quenching by dimethylformamide,
and deprotection by a trifluoroacetic acid treatment.
Key words: 2-hydroxy-6-methylbenzaldehyde; alarm pheromone; sex pheromone;
Astigmata; Acaridae
-26-
Increases in Hematopoietic Responses Caused by ภ-Glucans
in Mice
Tomoko Tateishi, Naohito Ohno, Yoshiyuki Adachi, and Toshiro Yadomae๕
Laboratory of Immunopharmacology of Microbial Products, School of Pharmacy, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo 192-03, Japan
Received March 25, 1997
@The effects of various (1จ3)-ภ-D-glucans on hematopoietic responses
of mice were investigated by measuring colony stimulating activity in sera
and ascites of the mice administered glucan. We have demonstrated that
the hematopoietic response was increased by various structures of (1จ3)-ภ-D-glucans,
i.e. soluble glucans (linear, branched, single helix, triple helix) and
particulate glucans. From the viewpoint of structure and activity relationships,
we found several characteristic features: i) hematopoietic response induced
by the particulate glucan disappeared faster than that by the soluble glucans,
ii) conformation of the glucans, single vs. triple helix, are relatively
independent of the response, iii) linear glucan had a weaker response,
and iv) there is a strong strain-dependency of the response. These results
corresponded well with the fact that branched (1จ3)-ภ-D-glucans, but
not linear and not particulate, are often used as biological response modifiers
for cancer patients.
Key words: ภ-glucan; colony stimulating factor; biological response modifier;
immunomodulator
-27-
Purification and Characterization of a Cysteine Protease
from Corms of Freesia, Freesia reflacta
Makoto Kaneda,๕ Hiroo Yonezawa, and Tetsuya Uchikoba
Laboratory of Biochemistry, Department of Chemistry, Faculty of Science, Kagoshima University, 1-21-35 Korimoto, Kagoshima 890, Japan
Received April 7, 1997
@A protease (freesia protease B) has been purified to electrophoretic
homogeneity from corms of freesia, Freesia reflacta by five steps of chromatography.
Its Mr was estimated to be about 26,000 by SDS-PAGE. The optimum pH of
the enzyme was 6.0-7.0 at 30C using casein as a substrate. The enzyme
was strongly inhibited by p-chloromercuribenzoic acid but not by phenylmethanesulphonylfluoride
and EDTA. These results indicate that freesia protease B is a cysteine
protease. Nine sites of oxidized insulin B-chain were cleaved by freesia
protease B in 24 h of hydrolysis. The four cleavage sites among them resembled
those of papain. From the digestion of five peptidyl substrates the specificity
of freesia protease B was found to be approximately broad, but the preferential
cleavage sites were negatively charged residues at P'(/)1 positions. Freesia
protease B preferred also the large hydrophobic amino acid residues at
the P2 position, in a similar manner to papain. The amino terminal sequence
of freesia protease B was identical with those of papain in regard to the
conservative residues of cysteine protease.
Key words: plant corm; cysteine protease; plant endopeptidase; freesia;
Freesia reflacta
-28-
Beta Ray-induced Scission of DNA in Tritiated Water and
Protection by a Green Tea Percolate and (-)-Epigallocatechin Gallate
Hiroe Yoshioka, Hiromu Kurosaki, Kouichi Yoshinaga,* Kieko Saito,** and Hisashi Yoshioka**
Radiochemistry Research Laboratory, and * Department of Chemistry, Faculty
of Science, Shizuoka University, 836 Ohya, Shizuoka-shi 422, Japan
** Graduate School of Nutritional and Environmental Sciences, University
of Shizuoka, 52-1 Yada, Shizuoka-shi 422, Japan
Received April 14, 1997
@The ภ-ray induced scission of puC18 plasmid DNA from E. coli in tritiated
water was examined in the presence or absence of a green tea percolate
(TP) and the main constituent, (-)-epigallocatechin gallate (EGCg). An
analysis of the ratio of the original closed-circular to the open-circular
form of DNA, which was formed by the strand scission of DNA, revealed that
TP and EGCg showed a protective effect on DNA scission depending on their
concentrations. A new technique, named solid state spin trapping, was applied
to examine this scavenging ability toward the hydroxyl (OH) radical generated
in tritiated water. The result was kinetically analyzed to reveal that
TP and EGCg showed the scavenging effect, suggesting that the protective
effect on DNA scission was attributable to the scavenging effect on the
OH radical.
Key words: beta-ray-induced DNA scission; green tea; (-)-epigallocatechin
gallate (EGCg); hydroxyl radical; spin trapping
-29-
Synthesis of (})-Hiburipyranone, a Cytotoxic Metabolite
of Marine Sponge Mycale adhaerens
Kanako Uchida, Hidenori Watanabe, and Kenji Mori ๕
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan
Received April 15, 1997
@Hiburipyranone, a cytotoxic metabolite of the marine sponge, Mycale adhaerens,
was synthesized as a racemate.
Key words: cytotoxin; hiburipyranone; isocoumarin derivative; Mycale adhaerens
-30-
Purification and Characterization of Novel Whey Glycoprotein
WGP-88 Which Binds to a Monoclonal Antibody to PAS-4 Glycoprotein in the
Bovine Milk Fat Globule Membrane
Sik Hwangbo,๕ Norihiro Azuma, Jun-ichi Kurisaki,* and Choemon Kanno ๕๕
Department of Applied Biochemistry, College of Agriculture, Utsunomiya
University, Utsunomiya 321, Japan
* Department of Animal Products, National Institute of Animal Industry,
Tsukuba 305, Japan
Received May 9, 1997
@A monoclonal antibody to the PAS-4 glycoprotein (78 kDa) of the bovine
milk fat globule membrane (MFGM) specifically recognized PAS-4, and was
named KAS4. A component recognized by KAS4 was found in whey protein, this
being a glycoprotein of 88 kDa by SDS-PAGE and named WGP-88. WGP-88 was
purified and characterized in comparison with PAS-4. WGP-88 had apparent
pI values of 6.45 and 6.39, while those of PAS-4 were 7.39 and 7.35. Neuraminidase
digestion shifted the pI values of WGP-88 to 6.57 and of PAS-4 to 7.52.
WGP-88 was rich in polar amino residues (44.9 mol%), while PAS-4 was abundant
in nonpolar amino acid residues (48.7 mol%). WGP-88 contained 17.1% of
carbohydrate and PAS-4 had 7.2%. The results of reductive hydrolysis, N-glycanase
digestion, and a lectin blot analysis suggested that N- and O-linked sugar
chains were contained in both glycoproteins. WGP-88 and PAS-4 had a different
N-terminal amino acid sequence. WGP-88 and PAS-4 respectively inhibited
competitively the binding of KAS4 to PAS-4 and WGP-88. Our studies revealed
WGP-88 recognized by KAS4 mAb to be a novel whey protein and to have different
biochemical properties from those of PAS-4.
Key words: milk whey glycoprotein; WGP-88; PAS-4; monoclonal antibody;
fat globule membrane
-31-
Note
Purification and Some Properties of Lignostilbene-ฟ,ภ-dioxygenase Isozyme
IV from Pseudomonas paucimobilis TMY1009
Shigehiro Kamoda,๕ Tamami Terada, and Yoshimasa Saburi
Department of Biomaterial Sciences, Graduate School of Agriculture and Agricultural Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan
Received December 2, 1996
@Lignostilbene-ฟ,ภ-dioxygenase isozyme IV was purified from ultrasonic
extracts of Pseudomonas paucimobilis TMY1009 through four steps of column
chromatography. The fraction obtained gave a single band on SDS-PAGE and
a single peak on reversed-phase HPLC and DEAE-HPLC. The specific activity
and Km of purified isozyme IV were 110 สkat/g and 4.2 สM for 4,4'-dihydroxy-3,3'-dimethoxystilbene,
150 สkat/g and 3.3 สM for 4,2'-dihydroxy-3,3'-dimethoxy-5'-(2''-carboxyvinyl)stilbene.
The molecular mass of intact isozyme IV was estimated to be 94 kDa by gel
permeation chromatography, and that of its subunits was 52 kDa by SDS-PAGE
under denaturing conditions. The N-terminal amino acid sequence of isozyme
IV differed slightly from that of other isozymes. Isozyme IV seemed to
be composed of two identical subunits, มม.
Key words: lignostilbene-ฟ,ภ-dioxygenase; lignin; Pseudomonas
-32-
Note
Orientation of Photosynthetic Reaction Center Reconstituted in Neutral
and Charged Liposomes
Masayuki Hara,*,**,๕ Takao Ueno,*** Takaaki Fujii,*** Qing Yang,**** Yasuo Asada,* and Jun Miyake,*,**
* National Institute of Bioscience and Human-Technology, AIST, MITI,
1-1 Higashi, Tsukuba, Ibaraki 305, Japan
** National Institute for Advanced and Interdisciplinary Research, AIST,
MITI, 1-1-4 Higashi, Tsukuba, Ibaraki 305, Japan
*** Department of Bioresources Chemistry, Chiba University, 648 Matsudo,
Matsudo-shi 271, Japan
**** Tsukuba Research Laboratry, Stanley Electric Co., Ltd., 5-9-5 Tokodai,
Tsukuba, Ibaraki 300-26, Japan
Received January 17, 1997
@The photosynthetic reaction center from the photosynthetic bacterium
Rhodobacter sphaeroides was reconstituted into neutral, positively charged,
or negatively charged liposomes. About 70% of photosynthtetic reaction
centers were reconstituted in the proteoliposomes exposing their H-subunit
outside with positively charged lipids while only 30-40% of them were in
the same topological orientation with neutral or negatively charged lipids.
Key words: photosynthetic reaction center; orientation; charged lipid;
liposomes; reconstitution
-33-
Note
Transient Expression of Goat Growth Hormone Gene in Poplar (Populus alba
L.) Protoplasts: A Quick Method for Detection of Foreign Gene Expression
in mRNA Level
Jingbo Qiao, Yasuyuki Ishihara, Hiroyuki Kuroda, Fukumi Sakai, Hiroshi Sakai,* and Tohru Komano*
Wood Research Institute, Kyoto University, Uji, Kyoto 611, Japan
* Laboratory of Biochemistry, Department of Agricultural Chemistry, Kyoto
University, Kyoto 606, Japan
Received January 22, 1997
@We developed a sensitive, accurate, and fast method to detect foreign
gene expression using reverse transcriptase-mediated polymerase chain reaction
(RT-PCR) in poplar tree (Populus alba L.) protoplasts. Template total RNA
was purified by removing the transfected foreign gene vector completely
with DNase I treatment before the RT reaction. Expression of cDNA that
encodes goat growth hormone was confirmed at the mRNA level 24 h after
electroporation mediated DNA transfer.
Key words: transient expression; RT-PCR; goat growth hormone gene; Populus
alba L.
-34-
Note
Polyamine Content of Ordinary Foodstuffs and Various Fermented Foods
Akiko Okamoto, Eriko Sugi, Yukimichi Koizumi, Fujiharu Yanagida, and Shigezo Udaka๕
Department of Brewing and Fermentation, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156, Japan
Received February 17, 1997
@Soybeans, tea leaves, and mushrooms were conspicuously rich in spermidine,
while oranges contained a large amount of putrescine. Among the fermented
foods, soy sauces were rich in putrescine and histamine, while Japanese
sake contained plenty of agmatine. These polyamines are thought to be produced
from amino acids during fermentation with amino acid decarboxylases formed
by the microorganisms.
Key words: polyamine; food polyamine; histamine; agmatine
-35-
Note
High Rate Production in Static Culture of Bacterial Cellulose from Sucrose
by a Newly Isolated Acetobacter Strain
Yukiko Kojima, Akira Seto, Naoto Tonouchi, Takayasu Tsuchida, and Fumihiro Yoshinaga
Bio-Polymer Research Co., Ltd., 3-2-1 Sakato, Takatsu-ku, Kawasaki 213, Japan
Received February 19, 1997
@For the industrial production of bacterial cellulose from sucrose in
static cultures, the possibility of a high rate of cellulose production
was investigated. An Acetobacter strain, S-35, which had been isolated
from a grape, was selected from 1500 isolates. This strain was found to
produce a large amount of cellulose from either glucose or fructose. Using
this strain, high cellulose production rates of 3.3 g/liter/d or 40 g/m2/d
from sucrose were seen in static culture.
Key words: Acetobacter; cellulose; static; production rate; sucrose
-36-
Note
Generation of Basidiomycetous Hyphal Cell-aggregates by Addition of the
Arg-Gly-Asp Motif-containing Fragment of High-molecular-weight Cell-adhesion
Protein MFBA Derived from the Basidiomycete Lentinus edodes
Toru Yasuda, Hiroki Ishihara,๕ Hitoshi Amano, and Kazuo Shishido๕๕
Department of Life Science, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama 226, Japan
Received February 26, 1997
@The Arg-Gly-Asp (RGD) motif-containing fragment of high-molecular-weight
cell-adhesion protein MFBA derived from Lentinus edodes caused a significant
aggregation of the fragmented hyphal cells of Schizophyllum commune. This
fungal cell-aggregation was inhibited by a previous treatment of the cells
with the Gly-Arg-Gly-Asp-Ser-Pro peptide, but not with the Gly-Arg-Gly-Glu-Ser-Pro
peptide, showing that the RGD motif is essential for the cell-aggregation
activity.
Key words: Arg-Gly-Asp (RGD) motif; hyphal cell-aggregation; Lentinus edodes
; Schizophyllum commune
-37-
Note
High Responsiveness to Thyroid Hormone of Adult Rat Primary Hepatocytes
Cultured on EHS-Gel
Hiroaki Oda,๕ Katsura Nozawa, Fuyuko Miyachi, Ai Shimizu, Yukiko Iwasaki, and Atsushi Kakinuma
Laboratory of Nutritional Biochemistry, Department of Applied Biological Sciences, Nagoya University, Nagoya 464-01, Japan
Received March 3, 1997
@The induction of malic enzyme gene expression by triiodothyronine and
insulin was severely blunted in rat monolayer hepatocytes cultured on type
I collagen compared with that in spherical hepatocytes cultured on a reconstituted
basement membrane gel (EHS-gel). Although the mRNA level of thyroid hormone
receptor ภ (TRภ) gradually decreased in the monolayer hepatocytes during
culture, the mRNA level in the hepatocytes on EHS-gel was maintained at
around the in vivo level. Our results suggest that the maintenance of TRภ
mRNA on EHS-gel is responsible for the high responsiveness to thyroid hormone
in a hepatocyte culture.
Key words: hepatocyte culture; thyroid hormone; extracellular matrix; TRภ
-38-
Note
Cloning of a Gene Encoding a Putative Xylanase with a Cellulose-Binding
Domain from Humicola grisea*
Hiroshi Iikura, Shou Takashima, Akira Nakamura, Haruhiko Masaki, and Takeshi Uozumi ๕
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan
Received March 5, 1997
@We have isolated a genomic clone of a putative xylanase gene (xyn1) from
Humicola grisea by using the DNA fragment encoding a cellulose-binding
domain of H. grisea cellobiohydrolase 1 as a probe. The translation product
of the xyn1 gene predicts a xylanase of 429 amino acids in length, with
a cellulose-binding domain in the C-terminus.
Key words: cellulose-binding domain; Humicola grisea; xylanase
-39-
Note
DNA Sequence of Bacillus subtilis (natto) NR-1 ม-Glutamyltranspeptidase
Gene, ggt
Yoshihiro Ogawa,๕ Dai Sugiura,* Hiroshi Motai, Katsumi Yuasa, and Yasutaka Tahara*
Research and Development Division, Kikkoman Corporation, 399 Noda, Noda-shi,
Chiba 278, Japan
* Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka
University, 836 Ohya, Shizuoka-shi, Shizuoka 422, Japan
Received March 6, 1997
@The ggt encoding ม-glutamyltranspeptidase (GGT) from Bacillus subtilis
(natto) was cloned and sequenced. The DNA sequence contains a single open
reading frame of 1761 bp that might be translated to a protein of 587 amino
acid residues, and indicates that B. subtilis (natto) GGT is synthesized
as prepro-GGT and processed later into large and small subunits. The putative
catabolite responsive element (CRE) was located upstream of the ggt coding
region.
Key words: Bacillus subtilis (natto); ม-glutamyltranspeptidase; cloning;
DNA; sequence
-40-
Note
Nitroguanidination of the Amino Groups of Ribonuclease T1 with N-Methyl-N'-nitro-
N-nitrosoguanidine๕
Kenji Takahashi
School of Life Science, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo 192-03, Japan
Received March 24, 1997
@Ribonuclease T1 was nitroguanidinated with N-methyl-N'-nitro-N-nitrosoguanidine
to investigate the effects of modification of the amino groups on the activity
and at the same time to shed some light on its usefulness in specific chemical
modification of proteins. Under the conditions used (at pH 7.0 and 37C
for 72 h), the amino group of the NH2-terminal Ala 1 was modified completely,
and the ร-amino group of Lys 41 about 90%. No other residue was modified.
The modified enzyme had approximately 84% of the original activity toward
RNA, and showed a characteristic absorption maximum at 270 nm.
Key words: amino groups; chemical modification; nitroguanidination; N-methyl-N'-nitro-N-nitrosoguanidine;
RNase T1
-41-
Note
Effects of Benadrostin, a Poly (ADP-Ribose) Polymerase Inhibitor, on Apoptosis
of HL-60 Induced by 5-Azacytidine
Hiroyasu Tobe,๕ Mitsune Yamaguchi, Tomoji Kocha, and Takaaki Aoyagi
Showa College of Pharmaceutical Sciences, 3165 Higashitamagawagakuen 3-chome, Machida-shi, Tokyo 194, Japan
Received April 8, 1997
@We studied the effects of benadrostin, an inhibitor of poly (ADP-ribose)
polymerase, on apoptosis of HL-60 cells induced by 5-azacytidine. Benadrostin
interfered with the apoptosis and inhibited the cell death in a dose- and
time-dependent manner.
Key words: apoptosis; 5-azacytidine; Zn ion; benadrostin; HL-60
-42-
Note
Identification of Cyanidin 3-O-(3'',6''-O-Dimalonyl-ภ-glucopyranoside)
as a Flower Pigment of Chrysanthemum (Dendranthema grandiflorum)
Masayoshi Nakayama,๕ Masaji Koshioka, Michio Shibata, Syuntaro Hiradate,* Hajime Sugie,* and Masa-atsu Yamaguchi**
National Research Institute of Vegetables, Ornmental Plants and Tea
(NIVOT), Ministry of Agriculture, Forestry, and Fisheries, 360 Kusawa,
Ano, Mie 514-23, Japan
* National Institute of Agro-Environmental Sciences, Ministry of Agriculture,
Forestry, and Fisheries, 3-1-1 Kannondai, Tsukuba, Ibaraki 305, Japan
** Department of Food Science and Technology, Minami-Kyushu University,
Takanabe, Miyazaki 884, Japan
Received April 14, 1997
@It had been suggested that cyanidin 3-O-(6''-O-monomalonyl-ภ-glucopyranoside)
and another unknown compound occur as major pigments in the purplish-red
flower of chrysanthemum (Dendranthema grandiflorum). We determined the
structure of the unknown compound as cyanidin 3-O-(3'',6''-O-dimalonyl-ภ-glucopyranoside)
by FAB-MS and 1H-NMR. This is the first report of the identification of
the cyanidin 3-dimalonyl glucoside as a flower pigment.
Key words: chrysanthemum (Dendranthema grandiflorum); purplish-red flower
pigment; acylated anthocyanin; cyanidin 3-O-(3'',6''-O-dimalonyl-ภ-glucopyranoside)
-43-
Rapid Paper
Protease Catalyzed Reaction of Polyamine Incorporation into
Protein Shunichi Suzuki,๕ Katsunori Kobayashi, Shigeru Yamanaka, and Kenzo Yokozeki
Central Research Laboratories, Ajinomoto Co., Inc., Suzuki-cho, Kawasaki-ku, Kawasaki, Kanagawa 210, Japan
Received April 30, 1997
@Radiolabeled putrescine incorporation into dimethyl casein is widely
measured in transglutaminase assays. We assessed the hypothesis that proteases
can catalyze this reaction. Acceleration of putrescine incorporation by
chymotrypsin, subtilisin, and papain, all purified to homogeneity, was
investigated, and all of the samples tested accelerated the reaction. These
effects disappeared when the proteases were first boiled. The rate of incorporation
was proportional to the enzyme concentration. The amount incorporated when
chymotrypsin was tested was proportional to the reaction time. On the basis
of these findings, we concluded that some proteases catalyze reactions
with putrescine incorporation.
Key words: chymotrypsin; subtilisin; papain; amine-incorporating activity;
screening for transglutaminase
-44-
Short Communication
A Ribosome-Inactivating Protein from Amaranthus viridis
Seok-Yoon Kwon, Chung Sun An,* Jang Ryol Liu, and Kyung-Hee Paek**,๕
Korea Research Institute of Bioscience and Biotechnology, P. O. Box
115, Taejon 305-333, Korea
* Department of Biology, Seoul National University, Seoul 151-742, Korea
** Graduate School of Biotechnology, Korea University, Seoul 136-701, Korea
Received February 3, 1997
@An antiviral protein purified from the leaves of Amaranthus viridis was
named amaranthin. The in vivo antiviral activity of amaranthin was confirmed
in tobacco mosaic virus (TMV) infection test on Nicotiana glutinosa leaves.
The molecular mass of the amaranthin was estimated about 30 kDa by SDS-PAGE
and the pI was measured as 9.8 by isoelectric focusing (IEF) analysis.
Cytotoxicity of the amaranthin using in vitro translation inhibition assay
was similar to that of pokeweed antiviral protein (PAP) with IC(/)50 of
25 pM. Depurination activity (N-glycosidase activity) against animal rRNA
was also confirmed.
Key words: antiviral protein; Amaranthus viridis; in vitro translation;
RIP; TMV
-45-
Short Communication
Synthesis of 2-O-Fucosyl Sulfatide, a Blocker of L- and P-Selectin๕
Hideharu Ishida, Satoshi Sago, Takao Ikami,* Makoto Kiso,๕๕ and Akira Hasegawa
Department of Applied Bioorganic Chemistry, Gifu University, Gifu 501-11,
Japan
* Drug Discovery Research Department, Sanwa Kagaku Kenkyusho Co., Ltd.,
363 Shiosaki, Hokusei-cho, Mie 511-04, Japan
Received May 19, 1997
@A new derivative of sulfatide, 2-O-ฟ-L-fucopyranosyl sulfatide, was
synthesized. The compound inhibited the binding of HL-60 cells, which express
sialyl Lewis X, to P- and L-selectin more than the corresponding non fucosylated
compound.
Key words: selectin blocker; sulfatide; inflammatory disease; cell adhesion
-46-
Short Communication
Distinguishable Action between Acid-stable and Neutral ฟ-Amylases from
Shochu Koji (Aspergillus kawachii )
Toshihiko Suganuma,๕ Naoyuki Noda, Hiroyuki Honbo, and Kanefumi Kitahara
Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890, Japan
Received June 9, 1997
@Acid-stable (KAA) and neutral (KNA) ฟ-amylases from shochu koji (A.
kawachii ) were purified and their actions towards maltooligosaccharides
were studied. KAA could be distinguished from KNA by the following actions:
with KAA, maltopentaose (G5) was preferentially hydrolyzed at the third
glycoside bond, and the addition of potassium thiocyanate (KSCN) decreased
the rate of CNP-release from 2-chloro-4-nitrophenyl-ฟ-maltotrioside (CNP-G3).
Key words: acid-stable ฟ-amylase; 2-chloro-4-nitrophenyl-ฟ-maltotrioside;
Aspergillus kawachii; cleavage pattern; maltooligosaccharide