(Vol.61 No.8 1997)
-Review-
Molecular Mechanism in -Glucosidase and Glucoamylase
Seiya Chiba 1233
Streptokinase Recovery by Cross-Flow Microfiltration: Study
of Enzyme Denaturation
Inmaculada Hernandez-pinzon, Francisco Millan, and Juan Bautista 1240
The Phylogeny of Acetic Acid Bacteria Based on the Partial
Sequences of 16S Ribosomal RNA: The Elevation of the Subgenus Gluconoacetobacter
to the Generic Level
Yuzo Yamada, Ken-ichiro Hoshino, and Tetsuya Ishikawa 1244
Screening for Fungi That Have High Lipolytic and Acidolytic
Activities in Biomass Support Particles
Tsunetoshi Miura and Tsuneo Yamane 1252
Enzymatic Synthesis of Oligosaccharides Containing Gal4Gal
Disaccharide at the Non-Reducing End Using -Galactanase from Penicillium
citrinum
Hiroshi Fujimoto, Hirofumi Nakano, Megumi Isomura, Sumio Kitahata,
and Katsumi Ajisaka 1258
Characterization of Hydroperoxides and Carbonyl Compounds
Obtained by Lipoxygenase Extracts of Selected Microorganisms
Barbara Bisakowski, Xavier Perraud, and Selim Kermasha 1262
Cloning and Characterization of the Gene Encoding a Novel
-Galactosidase from Bacillus circulans
Yoshiyuki Ito and Takashi Sasaki 1270
Substrate Specificity of Honeydew Melon Protease D, a Plant
Serine Endopeptidase
Hiroo Yonezawa, Tetsuya Uchikoba, and Mokoto Kaneda 1277
NMR Spectroscopic Analysis of Sulfated -1,3-Xylan and
Sulfation Stereochemistry
Tohru Yamagaki, Yohji Tsuji, Masaakira Maeda, and Hiroshi Nakanishi
1281
Purification and Characterization of the Acid Soluble
26-kDa Polypeptide from Soybean Seeds
Michiko Momma, Kazutomo Haraguchi, Masayoshi Saito, Koichi Chikuni,
and Kyuya Harada 1286
Inhibition of Tumor Necrosis Factor- Production by Orally
Administering a Perilla Leaf Extract
Hiroshi Ueda and Masatoshi Yamazaki 1292
Effect of Eluent Composition on the Distribution Coefficient
of Saccharides on to a Cation-exchange Resin in Sodium-ion Form
Shuji Adachi and Ryuichi Matsuno 1296
Induction after Administering Paraquat of Heme Oxygenase-1
and Heat Shock Protein 70 in the Liver of Senescence-accelerated Mice
Yukiko Nakanishi and Kyoden Yasumoto 1302
Analysis of Water Sorption Behavior of Native and Denatured
Proteins by Solution Thermodynamics
Hitomi Kumagai, Hirotaka Seto, Hidetoshi Sakurai, Kenji Ishii, and
Hitoshi Kumagai 1307
Protein Phosphorylation during Heading Time in Rice
Setsuko Komatsu and Bao-Sen Xia 1312
Caries-inducing Activity of the Hydrogenated Derivative
of an Isomaltooligosaccharide Mixture in Rats
Jun Tsunehiro, Takashi Matsukubo, Masao Shiota, and Yoshinori Takaesu
1317
Structure of the Gene Encoding Rat Renin Binding Protein
Saori Takahashi 1323
Effect of Vitamin B6 Deficiency on an Antibody Production
in Mice
Shoko Doke, Naoki Inagaki, Takashi Hayakawa, and Haruhito Tsuge 1331
Changes in the Number and Apoptosis of Epithelial Cells
in the Colorectum of Wheat Bran-fed Rats Soon after Administering 1,2-Dimethylhydrazine
Satoshi Ishizuka, Kei Sonoyama, and Takanori Kasai 1337
Synthesis of (+)-(1S,2S,5R,6S)-1-Hydroxysamin from L-(+)-Arabinose
Satoshi Yamauchi and Yoshiro Kinoshita 1342
Cloning and Sequencing of the Genes for N-Acetylglucosamine
Use that Construct Divergent Operons (nagE-nagAC) from Vibrio cholerae
Non-O1
Naoko Yamano, Noriyoshi Oura, Jingyu Wang, and Shizu Fujishima 1349
Glutathione-independent Formaldehyde Dehydrogenase from
Pseudomons putida: Survey of Functional Groups with Special Regard for
Cysteine Residues
Daisuke Tsuru, Naoko Oda, Yo Matsuo, Sara Ishikawa, Kiyoshi Ito, and
Tadashi Yoshimoto 1354
Long-term Effects of Water-soluble Corn Bran Hemicellulose
on Glucose Tolerance in Obese and Non-obese Patients: Improved Insulin
Sensitivity and Glucose Metabolism in Obese Subjects
Hiroyuki Hanai, Mutsuhiro Ikuma, Yoshihiko Sato, Takayuki Iida, Yoshisuke
Hosoda, Isao Matsushita, Atsuhiro Nogaki, Masami Yamada, and Eizo Kaneko
1358
Role of Gibberellins in the Thermoperiodic Regulation
of Stem Elongation in Dendranthema grandiflorum Tzvelev
Takaaki Nishijima, Mizuo Nonaka, Masaji Koshioka, Hiroshi Ikeda, Mitsuru
Douzono, Hiroko Yamazaki, and Lewis N. Mander 1362
-note-
Transformation System for Aspergillus oryzae with Double Auxotrophic Mutations,
niaD and sC
Osamu Yamada, Byung Rho Lee, and Katsuya Gomi 1367
NADPH-dependent Reduction of Ethyl Acetoacetate Coupled
with Ethanol Oxidation in Kloeckera magna
Tadashi Kometani, Yukiko Sakai, Hisae Urai, Hidefumi Yoshii, and Ryuichi
Matsuno 1370
Inhibition of Sulfur Oxidizing Activity by Nickel Ion
in Thiobacillus thiooxidans NB1-3 Isolated from the Corroded Concrete
Yasuo Nogami, Terunobu Maeda, Atsunori Negishi, and Tsuyoshi Sugio
1373
Oxidative Stability of Powdery Tridocosahexaenoin Included
in Cyclodextrin and Its Application to Fish Meal Paste
Hidefumi Yoshii, Takeshi Furuta, Kenichi Kawasaki, Hiroshi Hirano,
Yasuhiro Funatsu, Akira Toyomi, and Suguru Nakayama 1376
Molecular Cloning and Characterization of a cDNA Encoding
Putative Phospholipid Hydroperoxide Glutathione Peroxidase from Spinach
Manabu Sugimoto, Satoshi Furui, and Yukio Suzuki 1379
Subsite Affinities of -Glucosidases from Spinach Seeds
Satoshi Furui, Manabu Sugimoto, and Yukio Suzuki 1382
Gibberellin Transport from the Cotyledon to Plumule in
the Flowering of Pharbitis nil
Listyani Wijayanti, Masatomo Kobayashi, Shozo Fujioka, and Akira Sakurai
1384
O-Methylation of 2,6-Dimethoxy-4-methylphenol by Aspergillus
glaucus and Their Possible Contribution to Katsuobushi Flavor
Hajime Yamauchi and Mikiharu Doi 1386
Identification and NH(>22<)2-terminal Amino Acid
Sequences of DnaK and GroEL Homologues in Moderate Eubacterial Halophiles
Masao Tokunaga, Kenzo Matsuoka, and Hiroko Tokunaga 1388
Ester Synthesis by NAD+-dependent Dehydrogenation of Hemiacetal:
Production of Methyl Formate by Cells of Methylotrophic Yeasts
Agung Primanto Murdanoto, Yasuyoshi Sakai, Langkah Sembiring, Yoshiki
Tani, and Nobuo Kato 1391
Highly Stereoselective Synthesis of (E)-Substituted Allylsilanes
via the Still-Wittig Rearrangement
Kazushige Fujii, Osamu Hara, and Youji Sakagami 1394
4-O-Caffeoylshikimic and 4-O-(p-Coumaroyl)shikimic Acids
from the Dwarf Tree Fern, Dicksonia antarctica
Tamio Saito, Hisakazu Yamane, Noboru Murofushi, Nobutaka Takahashi,
and Bernard O. Phinney 1397
Lipid Peroxidation-derived Hepatotoxic Aldehyde, 4-Hydroxy-2-hexenal,
in Fish
Tadashi Sakai, Yoh-ichi Matsushita, Kazuhiro Sugamoto, and Koji Uchida
1399
-Rapid Paper-
Evidence for the Periodically Alternating Microfibrillar Structure of Bacterial
Cellulose
Yoshinori Hori, Kunihiko Watanabe, Yasushi Morinaga, and Fumihiro Yoshinaga
1401
-Short Communication-
Unique Inhibition of Miltpain, a New Cysteine Proteinase from the Milt
of Chum Salmon, by o-Phenanthroline, Phenanthrequinone, Phenazine, and
Acridine
Choko Kawabata and Eiji Ichishima 1405
Pradimicin, a Mannose-binding Antibiotic, Induced Carbohydrate-mediated
Apoptosis in U937 Cells
Toshikazu Oki, Yoko Yamazaki, Tamotsu Furumai, and Yasuhiro Igarashi
1408
Structural Analyses of Xyloglucan Heptasaccharide by the
Post-source Decay Fragment Method using Matrix-assisted Laser Desorption/Ionization
Time-of-Flight Mass Spectrometry
Tohru Yamagaki, Yasushi Mitsuishi, and Hiroshi Nakanishi 1411
Synthesis and Biological Activities of Indolactone-V,
the Lactone Analogue of the Tumor Promoter (-)-Indolactam-V
Yu Nakagawa, Kazuhiro Irie, Yoshimasa Nakamura, Hajime Ohigashi, and
Hideo Hayashi 1415
-1-
Review
Molecular Mechanism in -Glucosidase and Glucoamylase
Seiya Chiba
Department of Applied Bioscience, Faculty of Agriculture, Hokkaido University, Sapporo 060, Japan
@The hydrolysis of glucosidic linkage catalyzed by every carbohydrate-hydrolase
is a reaction in which the product retains ( or ) or inverts
( or ) the anomeric configuration of the substrate. -Glucosidase
and glucoamylase are essentially distinguished by releasing -glucose
and -glucose, respectively, from the common substrates having -glucosidic
linkage. The distinction in the substrate specificities of the two enzymes
was explained by the subsite affinities in their active sites. The amino
acid sequences of the regions containing the catalytic sites were compared
in -glucosidases and glucoamylases from various sources. -Glucosidases
were suggested to be grouped into two families by their primary structures.
The catalytic reaction mechanisms of carbohydrate-hydrolases were discussed
in the two significant models of a nucleophilic displacement mechanism
and an oxocarbenium ion intermediate mechanism.
Key words: -glucosidase; glucoamylase; subsite affinity; Pompe's disease;
d-glucal hydration
-2-
Streptokinase Recovery by Cross-Flow Microfiltration: Study
of Enzyme Denaturation
Inmaculada Hernandez-pinzon, Francisco Millan,* and Juan Bautista
Department of Biochemistry, Bromatology, and Toxicology, Faculty of
Pharmacy, University of Sevilla, 41012 Sevilla, Spain
* Instituto de la Grasa y sus Derivados, C.S.I.C. Sevilla, Spain
Received October 14, 1996
@During streptokinase (SK) recovery by cross-flow microfiltration (CFMF),
a loss of activity of 14.4% was observed, after the initial volume was
concentrated 8-fold (VCF=8.0); 51.5% of the activity was recovered in the
filtrate and 34.1% remained in the retentate. Immunological experiments
using polyclonal antibodies against SK have demonstrated that SK activity
loss during CFMF processes could be related to denaturation of SK, forming
molecules of lower or no activity. Accumulation of denatured SK in the
retentate suggests that denaturation could be the results of an aggregation
phenomenon (as has been demonstrated by light scattering and chromatographic
studies), leading to the formation of protein aggregates that are retained
by the microfiltration (MF) membrane, affecting the fouling phenomenon
and the concentration in the retentate of permeable molecules.
Key words: streptokinase; cross-flow-microfiltration; enzyme denaturation;
activity loss; Streptococcus equisimilis
-3-
The Phylogeny of Acetic Acid Bacteria Based on the Partial
Sequences of 16S Ribosomal RNA: The Elevation of the Subgenus Gluconoacetobacter
to the Generic Level
Yuzo Yamada,* Ken-ichiro Hoshino, and Tetsuya Ishikawa**
Laboratory of Applied Microbiology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422, Japan
Received November 18, 1996
@Thirty-six strains of acetic acid bacteria classified in the genera Acetobacter,
Gluconobacter, and Acidomonas were examined for their partial base sequences
in positions 1220 through 1375, 156 bases, of 16S rRNA. The strains of
the Q(/)10-equipped Gluconobacter species examined were divided into two
subgroups, which included the type strains of Gluconobacter oxydans, the
type species of the genus Gluconobacter, and of a second species, Gluconobacter
cerinus, respectively. The base differences numbered four between the two
type strains. The strains of the Q9-equipped species examined classified
in the type subgenus Acetobacter of the genus Acetobacter were not very
distant phylogenetically from those of the genus Gluconobacter. The calculated
number of base differences was 9-6 between the type strains of G. oxydans
and G. cerinus and the type strains of Acetobacter aceti and Acetobacter
pasteurianus. In contrast, the strains of the Q(/)10-equipped species examined
classified in the subgenus Gluconoacetobacter of the genus Acetobacter
were very distant phylogenetically from those of the Acetobacter and Gluconobacter
species mentioned above. The number of base differences was calculated
to be 14-8. Furthermore, the strains of the methanol-assimilating, Q(/)10-equipped
species of the genus Acidomonas examined were located in phylogenetically
isolated positions. The type strain of Acidomonas methanolica (Acetobacter
methanolicus), the type species of the genus Acidomonas, had 16-9 base
differences. The data obtained here indicated that the members of the subgenus
Gluconoacetobacter of the genus Acetobacter can be distinguished at the
generic level. The new genus Gluconoacetobacter was proposed with the type
species, Gluconoacetobacter liquefaciens, in recognition of the genus Acidomonas
along with the genera Acetobacter and Gluconobacter in the classification
of the acetic acid bacteria.
Key words: Gluconoacetobacter; acetic acid bacteria; Gluconoacetobacter
liquefaciens; partial base sequences of 16S rRNA; taxonomy
-4-
Screening for Fungi That Have High Lipolytic and Acidolytic
Activities in Biomass Support Particles
Tsunetoshi Miura and Tsuneo Yamane
Laboratory of Molecular Biotechnology, Department of Applied Biological Sciences, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya, Aichi 464-01, Japan
Received November 20, 1996
@Extensive screening for cell-bound lipase-producing filamentous fungi
suitable for immobilization in biomass support particles (BSPs) was done.
A total of 91 fungi were assayed for their lipolytic activity after cultivation
and immobilization in BSPs. In the cultivation, polypepton and oleic acid
were used as an organic nitrogen source and a lipase inducer, respectively.
Several species, such as Rhizopus species and Rhizomucor miehei, showed
higher cell-bound lipolytic activity than that of Rhizopus chinensis, which
is known to have considerable high cell-bound lipolytic activity from previous
work. For two strains of fungi (Rhizopus stolonifer, Rhizomucor miehei),
the cell-bound lipolytic activities of cells immobilized in BSPs did not
show strong dependence on polypepton concentration (10-60 g/liter), while
the extracellular lipolytic activities in the culture liquid increased
with the increase in the polypepton concentration. Thirteen fungi that
showed high cell-bound lipolytic activities were examined with respect
to their acidolytic activities using fish oil and caprylic acid in n-hexane.
There was a roughly proportional relationship between lipolytic and acidolytic
activities (the linear correlation coefficient, R, was 0.63). As the most
potent fungus having the highest cell-bound acidolytic activity in BSP,
Rhizomucor miehei IFO 9740 was selected.
Key words: immobilized fungi; biomass support particles; BSP; cell-bound
lipolytic activity; cell-bound acidolytic activity
-5-
Enzymatic Synthesis of Oligosaccharides Containing Gal4Gal
Disaccharide at the Non-Reducing End Using -Galactanase from Penicillium
citrinum
Hiroshi Fujimoto, Hirofumi Nakano,* Megumi Isomura, Sumio Kitahata,* and Katsumi Ajisaka
Meiji Institute of Health Science, Naruda Odawara 250, Japan,
* Osaka Municipal Technical Research Institute, Osaka 536, Japan
Received November 27, 1996
@The transglycosylation reaction was done with a -galactanase from Penicillium
citrinum. The regioselectivity in the transglycosylation reaction was studied
using soy bean arabinogalactan as a donor and mono- or disaccharide derivatives
containing -galactosyl residue as acceptors. We also synthesized oligosaccharides
containing Gal14Gal sequence such as Gal14Gal14Glc, Gal14Gal1
3GlcNAc, Gal14Gal14GlcNAc, Gal14Gal16GlcNAc, and Gal14Gal13GalNAc
for use in the total synthesis of complex sugar chains.
Key words: -galactanase; Penicillium citrinum; transglycosylation
-6-
Characterization of Hydroperoxides and Carbonyl Compounds
Obtained by Lipoxygenase Extracts of Selected Microorganisms
Barbara Bisakowski, Xavier Perraud, and Selim Kermasha
Department of Food Science and Agricultural Chemistry, McGill University, 21,111 Lakeshore, Ste. Anne de Bellevue, Quebec, Canada H9X 3V9
Received December 9, 1996
@Partially purified lipoxygenase (LOX) extracts were obtained from Fusarium
proliferatum, Fusarium oxysporum, Chlorella pyrenoidosa, and Saccharomyces
cerevisiae; the enzymatic extract of F. proliferatum showed the highest
LOX activity while those of F. oxysporum and S. cerevisiae demonstrated
only 27.8 and 16.5% of the activity at pH 8.0, and 61.2 and 9.7% of the
enzyme activity at pH 10.0, respectively. The lowest LOX activity was that
in the C. pyrenoidosa extract. The microbial enzymatic preparations were
assayed with linoleic acid as substrate, which was bioconverted into 9-
and 13-hydroperoxides (HPODEs) by all four extracts; in additon, the LOX
activity in the F. oxysporum extract produced the 10- and 12-HPODEs from
linoleic acid while that of the C. pyrenoidosa extract produced only the
10-HPODE. When assayed with the 9- and 13-HPODEs as substrates, the selected
microbial extracts had secondary enzyme activities, one of which produced
hexanal. The highest hexanal-producing activity was 1.51 and 1.39 nmol
hexanol/mg protein/min in the F. proliferatum and C. pyrenoidosa extracts,
respectively, while those of F. oxysporum and S. cerevisiae had approximately
15% of the HPODE-cleaving enzyme activity. The C. pyrenoidosa extract had
the highest proportion of pentanone, which was produced at only one-fourth
the concentration by the HPODE-cleaving enzyme activity in the three other
microbial enzymatic extracts.
Key words: lipoxygenase; linoleic acid; hydroperoxides; carbonyl compounds
-7-
Cloning and Characterization of the Gene Encoding a Novel
-Galactosidase from Bacillus circulans
Yoshiyuki Ito* and Takashi Sasaki
Laboratory of Lactic Acid Bacteria Research, Central Research Institute, Meiji Milk Products Co., Ltd., Naruda 540, Odawara 250, Japan
Received December 13, 1996
@A novel -galactosidase (-gal) gene was cloned from Bacillus circulans
ATCC 31382. The coding region was 1,758 bp and encoded a polypeptide of
586 amino acids with a deduced molecular mass of 66,888. Active staining
for -gal showed that B. circulans ATCC 31382 produced three -gal isozymes.
Two of these were detected in Biolacta N5 (Daiwakasei Co.), but the product
of this novel gene corresponded to the one not contained in Biolacta N5.
@The novel -gal showed the highest amino acid sequence identity (43.3%)
with a 13>14 galactosidase from Xanthomonas manihotis and was
also highly similar to -gals from animals, plants, and fungi. This suggests
an evolutionary relationship between this novel gene and those of eukaryotic
origins. One of the two B. circulans -gals, the nucleotide sequences
of which are available in the GenBank, was 20% identical to the novel -gal.
Other bacterial -gals showed little or no similarity.
@We propose that this novel -gal be called B. circulans -gal-3, and
the gene be called bgaC.
Key words: Bacillus circulans ; -galactosidase; gene cloning; bgaC
-8-
Substrate Specificity of Honeydew Melon Protease D, a Plant
Serine Endopeptidase
Hiroo Yonezawa, Tetsuya Uchikoba, and Makoto Kaneda
Department of Chemistry, Faculty of Science, Kagoshima University, Korimoto 1-21-35, Kagoshima 890, Japan
Received December 16, 1996
@The substrate specificity of honeydew melon (Cucumis melo var. inodorus
Naud) protease D was studied by the use of synthetic substrates and oligopeptides
derived from a protein hydrolyzate. The hydrolysis rates of succinyl-(L-Ala)(/)1-3-p-nitroanilide
(Suc-(Ala)(/)1-3-pNA) the hydrolysis rate progressively rose in proportion
to the increased chain length. Benzyloxycarbonyl-L-tyrosine p-nitrophenyl
ester (Z-Tyr-ONp) and benzoyl-L-tyrosine ethyl ester (Bz-Tyr-OEt) were
cleaved by honeydew melon protease D, but benzoyl-L-arginine p-nitroanilide
(Bz-Arg-pNA), benzyloxycarbonyl-L-lysine p-nitrophenyl ester (Z-Lys-ONp)
and tosyl-l-arginine methyl ester (Tos-Arg-OMe) were not hydrolyzed. Contrary
to the results obtained by using synthetic substrates, the carboxyl sides
of charged amino acid residues were preferentially cleaved by the enzyme
in the oligopeptide substrates. The substrates that had charged or polar
amino acids at P2 positions were not cleaved. On the other hand, the non-polar
amino acid or proline at P2 were favored for hydrolysis. The information
concerning the subsite of protease D was obtained and is useful for synthesis
of a good substrate. As it is distinct from molecular mass, the substrate
specificity of honeydew melon protease D is most analogous to cucumisin
[EC 3.4.21.25] among serine proteases from cucurbitaceous plants.
Key words: Cucumis melo; Cucurbitaceae; honeydew melon; plant protease;
serine protease
-9-
NMR Spectroscopic Analysis of Sulfated -1,3-Xylan and
Sulfation Stereochemistry
Tohru Yamagaki, Yohji Tsuji, Masaakira Maeda, and Hiroshi Nakanishi*
Department of Biochemistry, Saitama University, Urawa, Saitama 338,
Japan
* National Institute of Bioscience and Human Technology, AIST, 1-1 Higashi,
Tsukuba, Ibaraki 305, Japan
Received January 10, 1997
@A novel sulfated -1,3-xylan product was synthesized from algal cell
wall microfibril homoxylan by the N,N-dimethylformamide (DMF)-SO3 complex
sulfation method. Antithrombin activity appeared in this product was 6.5
times higher than that of standard heparin. From the results of 1H- and
13C-NMR spectroscopic analyses by DQF-COSY and HMQC and an infrared spectroscopic
analysis, it was revealed that the ordered structure of -1,3-xylan as
a triple helix had decayed and the resulting conformational changes had
been caused by the sulfation reaction. The sulfated positions on the C-4
hydroxyl groups of the xylose residues were determined from 13C-NMR chemical
shifts, and it was found that regioselective sulfation had occurred predominantly
with the C-4 secondary hydroxyl groups to produce a mono-substituent. Another
type of sulfation of -1,4-xylan that showed no regioselectivity is considered
to have been due to the different conformation of both xylans chains such
as the triple helix in -1,3-xylan and the double straight chain like
cellulose in -1,4-xylan. Therefore, the different type of regioselective
sulfation of -1,3- and -1,4-xylan was caused by the difference in steric
hindrance due to these conformations. These different types of regioselective
sulfation with different linkage positions are also discussed for the secondary
hydroxyl groups in -1,3- and -1,4-glucan after chemoselective sulfation
of the C-6 primary hydroxyl groups.
Key words: Caulerpa cell wall -1,3-xylan; regioselective sulfation of
the secondary hydroxyl groups;
NMR spectroscopy; steric hindrance
-10-
Purification and Characterization of the Acid Soluble
26-kDa Polypeptide from Soybean Seeds
Michiko Momma, Kazutomo Haraguchi, Masayoshi Saito, Koichi Chikuni, and Kyuya Harada*
National Food Research Institute, Ministry of Agriculture Forestry,
and Fisheries, 2-1-2 Kannondai, Tsukuba, Ibaraki 305, Japan *
Faculty of Horticulture, Chiba University, 648 Matsudo, Chiba 271, Japan
Received January 13, 1997
@Whey proteins from soybean seeds of Japanese varieties were analyzed
by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Among 11 varieties
of soybean, three green and one black soybeans lacked a 26-kDa band that
was found in all yellow soybeans. In this paper, the 26-kDa protein was
named AS26k (acid soluble 26-kDa protein) temporarily. The AS26k protein
was purified from Glycine max cv. Nattosyoryu, which is yellow soybean,
through four purification steps: 30-35% saturated ammonium sulfate fractionation,
ion exchange chromatography on S Sepharose Fast Flow, gel filtration on
Sephadex G-100, and hydrophobic chromatography on phenyl Sepharose CL-4B.
Purified AS26k was cleaved with V8 proteinase from Staphylococcus aureus
or CNBr. The cleaved polypeptide contained two typical dehydrin motif sequences:
DEYGNPV and (M)DKIKEKLPG, and a 19 amino acids sequence similar to a pea
dehydrin. Native AS26k had a molecular mass of 32 kDa on gel filtration
and a pI of 7.2 on two-dimensional PAGE. Similarly to other dehydrins and
late embryogenesis abundant (LEA) proteins, AS26k was rich in hydrophilic
amino acids, and highly heat stable. These results showed that AS26k was
a dehydrin, a group II LEA protein in soybean seeds.
Key words: amino acid sequence; dehydrin/group II LEA protein; protein
purification; soybean; seed protein
-11-
Inhibition of Tumor Necrosis Factor- Production by Orally
Administering a Perilla Leaf Extract
Hiroshi Ueda and Masatoshi Yamazaki
Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko-machi, Kanagawa 199-01, Japan
Received January 13, 1997
@The overproduction of tumor necrosis factor- (TNF-) was suppressed
by orally administering a perilla leaf extract (PLE). When mice were successively
injected with OK-432, severe TNF- was induced in the serum, but this
elevated TNF- level was reduced after an oral administration of PLE (400
l/mouse). Oral administration of PLE also inhibited TNF- production
that was induced by muramyl dipeptide (500 g/mouse) and OK-432 (3 KE/mouse).
These characteristics were obtained from all strains of perilla. The inhibitory
activity against TNF- production was heat-stable, and the existence of
several active molecules was suggested. When PLE was passed through an
ultrafilter, the inhibitory activity against TNF- production was collected
in those fractions with a mass of 0.5 to 1 kDa and more than 10 kDa. When
PLE was solvent-extracted, the strongest activity was recognized with aqueous
preparation, although significant activity was also detected in preparations
extracted with n-hexane and ethyl acetate. These findings suggest that
the daily use of certain functional foods may be useful for controlling
the host defense system.
Key words: tumor necrosis factor-; Perilla frutescens
-12-
Effect of Eluent Composition on the Distribution Coefficient
of Saccharides on to a Cation-exchange Resin in Sodium-ion Form
Shuji Adachi and Ryuichi Matsuno
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan
Received January 14, 1997
@The distribution coefficients of glucose, maltose, and maltotriose on
to a cation-exchange resin in sodium-ion form were measured by using methanol-
or ethanol-water mixtures of various content as eluents. With increasing
methanol or ethanol content, the resin shrank and the coefficients increased.
This increase in the coefficients could not be explained by only the effect
of the swelling pressure of the resin on the distribution, and it seems
that the complex formation of Na+ and the saccharides played an important
role. The binding constants between some saccharides and Na+ were evaluated,
and the electronegativity of each saccharide relative to that of glucose
was estimated.
Key words: distribution coefficient; ion-exchange resin; saccharides; swelling
pressure; binding constant
-13-
Induction after Administering Paraquat of Heme Oxygenase-1
and Heat Shock Protein 70 in the Liver of Senescence-accelerated Mice
Yukiko Nakanishi and Kyoden Yasumoto
Research Institute for Food Science, Kyoto University, Gokasho, Uji, Kyoto 611, Japan
Received January 17, 1997
@Age-associated changes in the induction of heme oxygenase (HO-1) and
heat shock protein 70 (HSP70) after the administration of paraquat were
investigated in the liver of senescence-accelerated mice (SAMs). The extent
of HO-1 and HSP70 induced in response to paraquat decreased significantly
with age, and the level of oxidized proteins increased with age. Moreover,
the extent of induced HSP70 was lower in mice that were prone to accelerated
senescence (SAMP1 // Fky) than in mice that were resistant to accelerated
senescence (SAMR1/Fky) of the same age. These results suggest that an age-associated
decline in the levels of HO-1 and HSP70 enhanced oxidative damage during
the aging process. Age-dependent changes in HO-1 and in the levels of oxidized
proteins were examined in SAMP1 // Fky. The accumulation of oxidized proteins
was suppressed when HO-1 was induced, but increased markedly after the
induction of HO-1 decreased. Free iron, the residuum from heme degradation,
might mediate free radical production. The role of HO-1 is discussed in
relation to the oxidative damage associated with age.
Key words: age; senescence-accelerated mouse; heat shock protein 70; heme
oxygenase; stress
-14-
Analysis of Water Sorption Behavior of Native and Denatured
Proteins by Solution Thermodynamics
Hitomi Kumagai, Hirotaka Seto, Hidetoshi Sakurai, Kenji Ishii, and Hitoshi Kumagai*
Department of Agricultural and Biological Chemistry, College of Bioresource
Sciences, Nihon University, 3-34-1 Shimouma, Setagaya-ku, Tokyo 154, Japan
* Department of Applied Biological Chemistry, Division of Agriculture and
Agricultural Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku,
Tokyo 113, Japan
Received January 22, 1997
@Water sorption isotherms of native and denatured ovalbumin samples were
measured, and their affinity for water was quantitatively analyzed by solution
thermodynamics. The structural change of the proteins was evaluated by
several methods. Water activity above 0.9 was measured by adding a specific
amount of water to a sample, while that below 0.9 was measured with apparatus
for water sorption isotherms. Thus, water sorption isotherms for native
and denatured ovalbumin samples were obtained up to a water activity of
0.99. The water sorption behavior of ovalbumin was affected not by its
hydrophobicity but by its prominent conformational change. It was confirmed
that parameter G s calculated by solution thermodynamics was more suitable
for evaluating the affinity for water from water sorption isotherms than
G s(/)w calculated by conventional adsorption thermodynamics.
Key words: water sorption isotherm; solution thermodynamics; Gibbs free
energy; protein denaturation; ovalbumin
-15-
Protein Phosphorylation during Heading Time in Rice
Setsuko Komatsu and Bao-Sen Xia
Department of Molecular Biology, National Institute of Agrobiological Resources, Kannondai 2-1-2, Tsukuba, Ibaraki 305, Japan
Received January 28, 1997
@Protein phosphorylation may be required for plant cell response to phytohormones
and other extracellular signals. Protein phosphorylation and protein kinase
activity in the culm of heading time of rice (Oryza sativa L.) were studied.
Before heading, protein kinase activity was increased by Ca2+ in the membrane
fraction of the panicle and culm. The protein kinases with Mr of 51,900,
49,200, and 45,500 isolated from the membrane fraction of culm increased
the protein phosphorylation of Mr and pI of 40,000/7.5 and 40,000/7.6 in
the culm extract. The activation of protein kinases, associated with membrane
and subsequent protein phosphorylation, thus appears to be involved in
the regulation of heading time in rice.
Key words: Oryza sativa; Graminaceae; protein kinase; protein phosphorylation;
heading time
-16-
Caries-inducing Activity of the Hydrogenated Derivative
of an Isomaltooligosaccharide Mixture in Rats
Jun Tsunehiro, Takashi Matsukubo,* Masao Shiota,** and Yoshinori Takaesu *
Research and Development Center, Showa Sangyo Co., Ltd., Hinode, Funabashi,
Chiba 273, Japan
* Department of Hygiene and Community Dentistry, Tokyo Dental College,
Masago, Mihama-ku, Chiba 261, Japan
**Technical Division, Showa Sangyo Co., Ltd., Higashi Fukashiba, Kamisu,
Kashima-gun, Ibaraki 314-02, Japan
Received January 29, 1997
@The caries-inducing activity of the hydrogenated derivative of an isomaltooligosaccharide
mixture (IMO-H) was evaluated in vitro for its acidogenicity and in vivo
an experimental caries system with specific-pathogen-free (SPF) rats. Streptococcus
sobrinus 6715 (serotype g) did not produce a significant amount of acid
from IMO-H, whereas Streptococcus mutans MT8148 (serotype c) gradually
produced a small amount of acid, although the degree was less than that
of sucrose. In vivo experiments were conducted on rats which were provided
with the test sugars at two different times: at the time of organism inoculation,
and after the organisms had become completely established. IMO-H did not
induce significant dental caries in rats infected with the S. sobrinus
6715 or S. mutans MT8148R strain.
Key words: cariogenicity; hydrogenated isomaltooligosaccharide; sugar substitute;
mutans streptococci
-17-
Structure of the Gene Encoding Rat Renin Binding Protein
Saori Takahashi*
Department of Applied Biology, Faculty of Textile Science, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606, Japan
Received January 30, 1997
@The gene encoding renin-binding protein was isolated from a Sprague-Dawley
rat genomic DNA library and the exon-intron boundaries and transcription
initiation sites were identified. The gene was found to span about 9.0
kilobase pairs and to be composed of eleven exons. The exon size ranged
76 to 225 base pairs. All the exon-intron junctional sequences conformed
to the canonical GT-AG rule. The first exon encoded a 5'-untranslated region
with the ATG translation start codon being in exon 2. The promoter region
did not contain a TATA box, but it had two GC boxes in the 5'-untranslated
sequence.
The overall exon pattern of the rat RnBP gene correlated with the human
RnBP gene [S. Takahashi, H. Inoue, and Y. Miyake, J. Biol. Chem., 267,
13007-13013 (1992)]. Moreover, the sizes of exons 2 to 10 in the rat RnBP
gene were the same as those of the human RnBP gene. These results indicate
that the structure of the RnBP gene is conserved among animals.
Key words: renin; binding protein; high molecular weight; gene structure
-18-
Cysteine-S-conjugate -Lyase Activity and Pyridoxal Phosphate
Binding Site of Onion Alliin Lyase
Nobuko Kitamura, Naoya Shimomura, Junji Iseki, Mamoru Honma, Seiya Chiba, Satoshi Tahara, and Junya Mizutani
Faculty of Agriculture, Hokkaido University, Sapporo 060, Japan
Received February 3, 1997
@Purification of onion alliin lyase gave two fractions by cation exchange
chromatography. Both fractions showed the comparable high catalytic activity
of cysteine-S-conjugate -lyase with that of alliin lyase using S-(2-chloro-6-nitrophenyl)-L-cysteine
and alliin, S-allyl-L-cysteine sulfoxide as substrates. All the active
substrates tested with onion alliin lyase were also active to the cysteine-S-conjugate
-lyase of Mucor javanicus, but the catalytic activity of the Mucor enzyme
was lower for all the substrates. The pyridoxal phosphate binding site
of the onion alliin lyase was identified as Lys 285 in the amino acid sequence
deduced from cDNA which has been reported. This lysine was conserved in
all the sequences from the alliin lyase cDNAs, while similarity was not
found between the sequences around pyridoxal phosphate binding sites of
both the onion alliin lyase and the Mucor cysteine-S-conjugate -lyase.
Key words: onion alliin lyase; pyridoxal phosphate binding site; cysteine-S-conjugate
-lyase; S-chloronitrophenyl-l-cysteine; Mucor javanicus
-19-
Effect of Vitamin B6 Deficiency on an Antibody Production
in Mice
Shoko Doke,*,*** Naoki Inagaki,** Takashi Hayakawa,*** and Haruhito Tsuge***,
* Gifu City Women's College, 2693 Nagarafukumitu, Gifu 502, Japan
** Faculty of Pharmacy, Gifu Pharmaceutical University, 5-6-1 Mitahorahigashi,
Gifu 502, Japan
*** The United Graduate School of Agricultural Science, Gifu University,
1-1 Yanagido, Gifu 501-11, Japan
Received February 5, 1997
@To investigate the effects of vitamin B6 (B6) deficiency on an antibody
production in BALB/c mice, the production of specific immunoglobulin (Ig)
E antibody against dinitrophenylated ovalbumin (DNP-OVA) were measured
by the methods of enzyme linked immunosorbent assay. The mice fed on on
a B6 deficient diet for 4 weeks were immunized intraperitoneally with DNP-OVA
absorbed to aluminum hydroxide gel. The contents of anti DNP-IgE antibodies
in sera of B6 deficient mice significantly increased compared to that of
control mice fed on a diet containing B6. In addition, Interleukin-4, which
was known to induce IgE production in allergic reactions from splenocytes
of B6 deficient mice, was approximately four-fold higher than that in control
mice.
@According to the recovery test to the B6 deficient mice, that is feeding
the control diet for 21 days, all values in terms of the body, thymus,
and spleen weight, total serum protein, IgG, and anti DNP-IgE content,
regained almost the same levels as those of control. These results suggest
that B6 deficiency in mice would have relation to the stimulation of specific
IgE antibody production against DNP-OVA.
Key words: vitamin B6 deficiency; IgE antibody; ELISA; IL-4
-20-
Changes in the Number and Apoptosis of Epithelial Cells
in the Colorectum of Wheat Bran-fed Rats Soon after Administering 1,2-Dimethylhydrazine
Satoshi Ishizuka, Kei Sonoyama, and Takanori Kasai
Department of Bioscience and Chemistry, Faculty of Agriculture, Hokkaido University, Kita-9, Nishi-9, Kita-ku, Sapporo 060, Japan
Received February 5, 1997
@We investigated the effect of dietary wheat bran (Wb) on colonic tumorigenesis
soon after a single administration of 1,2-dimethylhydrazine (DMH). Rats
that had been fed on either a fiber-free diet or a 20% Wb diet were injected
with 1,2-dimethylhydrazine (20 mg/kg body weight). At 6 h, 12 h, 1 d, 3
d, or 7 d after the injection, the colorectum was excised for histological
analyses. The number of crypt cells more rapidly recovered in the 20% Wb
group than in the fiber-free group after its temporary reduction by injection
of DMH. At 6 h after the DMH treatment, the apoptotic cells were significantly
greater in number in the fiber-free group than in the 20% Wb group. In
contrast, those in distal colon were significantly fewer in the fiber-free
group than in the 20% Wb group at 7 d after the treatment. These results
suggest that the ingestion of Wb affected the turnover of colonic epithelial
cells and would thereby bring about a protective effect against DMH-induced
tumorigenesis.
Key words: apoptosis; 1,2-dimethylhydrazine; short-chain fatty acids; rat;
wheat bran
-21-
Synthesis of (+)-(1S,2S,5R,6S)-1-Hydroxysamin from L-(+)-Arabinose
Satoshi Yamauchi and Yoshiro Kinoshita
College of Agriculture, Ehime University, Tarumi 3-5-7, Matsuyama, Ehime 790, Japan
Received February 13, 1997
@As a model for the synthesis of optically active 6-aryl-2-aryloxy-1-hydroxy-3,7-dioxabicyclo[3.3.0]octanes,
which are 1,2-dioxygenated furofuran lignans, the most important intermediate,
(+)-(1S,2S,5R,6S)-1-hydroxysamin (1), was synthesized from L-(+)-arabinose.
Key words: samin; lignan; furofuran lignan
-22-
Cloning and Sequencing of the Genes for N-Acetylglucosamine
Use That Construct Divergent Operons (nagE-nagAC) from Vibrio cholerae
Non-O1
Naoko Yamano, Noriyoshi Oura, Jingyu Wang, and Shizu Fujishima
Osaka National Research Institute, Agency of Industrial Science and Technology, Ikeda, Osaka 563, Japan
Received February 13, 1997
@A 7.2-kb genomic DNA fragment containing N-acetylglucosamine-6-phosphate
deacetylase gene (nagA) was cloned from the chitinase-producing bacterium
Vibrio cholerae non-O1 strain 1148A (IFO 15429). Sequence analysis of the
DNA fragment found three other complete open reading frames (ORFs) and
the 5' end of an ORF. Amino acid sequences of two ORFs, ORF2 and ORF4,
showed similarity with that of NagC, the repressor of nag operons and that
of NagE, N-acetylglucosamine-specific transporter IINag of phosphoenolpyruvate
transport system of Escherichia coli, respectively. In the presence of
N-acetylglucosamine, nagA and ORF2 (nagC) were co-transcribed. ORF4 (nagE),
which is upstream from nagAC but is expressed in the opposite direction
was also transcriptionally induced in the presence of N-acetylglucosamine.
These results indicated that nagE-nagAC existed as divergent operons in
V. cholerae non-O1. Unlike E. coli, nagB and nagD were not in the operons.
Key words: N-acetylglucosamine-6-phosphate deacetylase; Escherichia coli;
sequence homology
-23-
Glutathione-independent Formaldehyde Dehydrogenase from
Pseudomons putida: Survey of Functional Groups with Special Regard for
Cysteine Residues
Daisuke Tsuru, Naoko Oda, Yo Matsuo,*, Sara Ishikawa,** Kiyoshi Ito,** and Tadashi Yoshimoto**
Department of Applied Microbial Technology, Kumamoto Institute of Technology,
4-22-1 Ikeda, Kumamoto 860, Japan
* Institute for Social Information Science, Fujitsu Laboratories Ltd.,
Mihama-ku, 1-9-3 Nakase, Chiba 161, Japan
**School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-machi,
Nagasaki 852, Japan
Received February 20, 1997
@The role of cysteine residues for structure and function of formaldehyde
dehydrogenase from Pseudomonas putida was analysed by amino acid sequence
comparison, homology-based structure modeling, site-directed mutagenesis,
and chemical modification. Five out of seven cysteine residues found in
the enzyme were concluded to coordinate with an active site zinc (Cys-46)
and structural zinc atoms (Cys-97, -100, -103, and -111) from the sequence
comparison with other Zn-containing medium-chain alcohol dehydrogenase
homologues. The three-dimensional structure model based on the known structure
of the horse liver E-type alcohol dehydrogenase (ADH) indicated that Cys-257
is located very far from the active site Zn and NAD+ binding region, suggesting
that Cys-257 does not participate in the enzyme reaction. The structure
also suggested that Cys-166 does not coordinate to active site Zn, but
Asp-169 functions as a Zn-ligand, instead.
Key words: alcohol dehydrogenase; glutathione-independent formaldehyde
dehydrogenase; homology modeling; Pseudomonas putida;
Zn-containing medium-chain alcohol dehydrogenase
-24-
Long-term Effects of Water-soluble Corn Bran Hemicellulose
on Glucose Tolerance in Obese and Non-obese Patients: Improved Insulin
Sensitivity and Glucose Metabolism in Obese Subjects
Hiroyuki Hanai, Mutsuhiro Ikuma, Yoshihiko Sato, Takayuki Iida, Yoshisuke Hosoda, Isao Matsushita, Atsuhiro Nogaki, Masami Yamada, and Eizo Kaneko
First Department of Medicine, Hamamatsu University School of Medicine, 3600 Handa-cho, Hamamatsu 431-31, Japan
Received February 27, 1997
@We examined the effect of soluble corn bran hemicellulose (CBH, 10 g/day)
on glucose control and serum insulin in three groups: patients with impaired
glucose tolerance (IGT) with (20 subjects) or without (8 subjects) obesity
and with healthy non-obese controls (10 subjects). Long-term supplementation
(6 months) with CBH decreased the post oGTT curve for patients with impaired
mild Type II diabetes, but not that for the controls. Hemoglobin A1c decreased
significantly during CBH supplementation in the obese patients, while the
fasting glucose level decreased in all three groups, although not significantly.
A decreased serum insulin response by oGTT was found in those patients
with IGT.
@The improved oGTT result was associated with improved insulin release
and perhaps with peripheral insulin sensitivity. These findings suggest
that CBH at a low dose might contribute to glycemic control and would play
a useful role in treating Type II diabetes patients.
Key words: insulin resistance; obesity; dietary fiber
-25-
Role of Gibberellins in the Thermoperiodic Regulation
of Stem Elongation in Dendranthema grandiflorum Tzvelev
Takaaki Nishijima, Mizuo Nonaka,* Masaji Koshioka, Hiroshi Ikeda, Mitsuru Douzono, Hiroko Yamazaki, and Lewis N. Mander**
National Research Institute of Vegetables, Ornamental Plants and Tea,
360 Kusawa, Ano-cho, Age-gun, Mie 514-23, Japan
* Kurume Branch, National Research Institute of Vegetables, Ornamental
Plants and Tea, Mii-cho 1823, Kurume-shi, Fukuoka 839, Japan
** Research School of Chemistry, The Australian National University, G.P.O.
Box 4, Canberra A.C.T. 2601, Australia
Received March 3, 1997
@Role of endogenous gibberellins (GAs) in the thermoperiodic regulation
of stem elongation in Dendranthema grandiflorum was investigated by gas
chromatography-mass spectrometry with deuterated GAs as internal standards.
GA1, GA20, GA19, GA44, and GA53, which all belong to the early 13-hydroxylation
pathway of GA-biosynthesis, and GA9, which belongs to the early nonhydroxylation
pathway, were identified from the stem. The thermoperiodic treatments employed
were a higher day temperature than night temperature (DT>NT) and several
hours of temperature drop from beginning of the day (TD). TD markedly decreased
the stem elongation rate when compared to the effect of DT>NT. Further,
TD decreased the concentrations of stem GA1, GA20, and GA19 more than DT>NT,
while TD increased the concentration of stem GA44 more than DT>NT. Thus,
TD probably retarded the conversion of GA44 to GA19, resulting in a low
concentration of GA1, the probable biologically-active GA. These changes
in endogenous GA concentration are probably an important physiological
factor for the thermoperiodic regulation of stem elongation in D. grandiflorum.
Key words: Dendranthema grandiflorum; DIF; temperature drop; gibberellin;
stem elongation
-26-
Note
Transformation System for Aspergillus oryzae with Double Auxotrophic Mutations,
niaD and sC
Osamu Yamada, Byung Rho Lee, and Katsuya Gomi
National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima 739, Japan
Received November 13, 1996
@We developed a transformation system for Aspergillus oryzae using the
Aspergillus nidulans sC gene encoding ATP sulfurylase as a selectable marker.
The sC- mutants can be readily isolated by positive selection for selenate
resistance, thereby the niaD- mutant strain of A. oryzae was bestowed with
the sC- mutation. Transformation of the A. oryzae host (niaD-, sC-) with
the plasmid carrying A. nidulans sC gave random and multi-copy integrants,
while that with the A. oryzae niaD-carrying plasmid occurred mainly by
single-copy and homologous integration events (more than 50% frequency),
indicating that with this transformation system, the transformation marker
could be selected according to the integration pattern one desires.
Key words: Aspergillus oryzae; transformation; ATP sulfurylase; nitrate
reductase
-27-
Note
NADPH-dependent Reduction of Ethyl Acetoacetate Coupled with Ethanol Oxidation
in Kloeckera magna
Tadashi Kometani, Yukiko Sakai, Hisae Urai, Hidefumi Yoshii, and Ryuichi Matsuno*
Department of Chemical and Biochemical Engineering, Toyama National
College of Technology, Hongo 13, Toyama 939, Japan
* Department of Food Science and Technology, Faculty of Agriculture, Kyoto
University, Kyoto 606, Japan
Received November 18, 1996
@We evaluated the catalytic ability of 29 yeast strains to reduce ethyl
acetoacetate (EA) in the presence of ethanol or glucose. In 18 yeast strains,
the reduction in the presence of ethanol proceeded as well as in the presence
of glucose. Among them, Kloeckera magna (AKU 4704) effectively catalyzed
the NADPH-dependent reduction of EA in the presence of ethanol. In this
reduction, 1 mol of EA was reduced by consuming 1 mol of ethanol. We found
that the NADPH regeneration system responsible for EA reduction in K. magna
was coupled with the oxidation of acetaldehyde to acetic acid catalyzed
by an NADP+-dependent aldehyde dehydrogenase.
Key words: Kloeckera magna; asymmetric reduction;
NADPH regeneration; ethanol; aldehyde dehydrogenase
-28-
Note
Inhibition of Sulfur Oxidizing Activity by Nickel Ion in Thiobacillus thiooxidans
NB1-3 Isolated from the Corroded Concrete
Yasuo Nogami, Terunobu Maeda,* Atsunori Negishi,* and Tsuyoshi Sugio
Department of Biological Function and Genetic Resources Science, Faculty
of Agriculture, Okayama University, 1-1-1 Tsushima Naka, Okayama 700, Japan
* Hazama Corporation, Technical Research Institute, 515-1 Nishimukai Karima
Tsukuba, Japan
Received December 16, 1996
@It is of great importance to find ways to protect concrete from corrosion,
to maintain the concrete structure for a long time. A sulfur-oxidizing
bacterium, Thiobacillus thiooxidans, has been known to play a crucial role
in concrete corrosion and the concrete supplemented with nickel was resistant
to corrosion. To obtain biological bases of this Ni protection, the effects
of Ni on sulfur dioxygenase and sulfite oxidase of T. thiooxidans NB1-3
isolated from corroded concrete were studied. Nickel sulfate strongly inhibited
a sulfur dioxygenase that catalyzes the oxidation of elemental sulfur to
shlfite and a sulfite oxidase that catalyzes oxidation of sulfite to sulfate.
Nickel sulfate, antimycin A, 2-n-heptyl-4-hydroxyquinoline-N-oxide, and
myxothiazol inhibited both sulfite oxidase and ubiquinol oxidase. Reduced
mammalian cytochrome c was not oxidized by a cell extract of strain NB1-3.
The b and a-type cytochromes in the plasma membrane was reduced by sulfite
and ubiquinol-2 and these reductions were inhibited by NiSO4. The amounts
of Ni in the plasma membrane with or without 0.1 mM nickel treatment were
32.6 and 2.8 nmol/mg protein, respectively. These results indicate that
nickel binds to the plasma membrane and inhibits sulfur dioxygenase and
sulfite oxidase, and as a result, inhibits cell growth.
Key words: sulfur-oxidizing bacterium;
Thiobacillus thiooxidans; concrete; corrosion; nickel
-29-
Note
Oxidative Stability of Powdery Tridocosahexaenoin Included in Cyclodextrin
and Its Application to Fish Meal Paste
Hidefumi Yoshii, Takeshi Furuta,*, Kenichi Kawasaki,** Hiroshi Hirano,** Yasuhiro Funatsu,** Akira Toyomi,*** and Suguru Nakayama****
Department of Biochemical Engineering, Toyama National College of Technology,
Toyama 939, Japan
* Department of Biotechnology, Tottori University, Tottori 680, Japan
** Food Research Lab., Toyama Food Research Institute, Toyama 939, Japan
*** Kurimoto Co., Ltd., Osaka 559, Japan
**** Food Research Lab., Central Research Institute, Maruha Coorporation,
16-2 Wadai, Tsukuba 300-2, Japan
Received January 16, 1997
@The inclusion complex between DHA oil containing tridocosahexaenoin to
45% and cyclodextrin (CD) was prepared by a twin-screw kneader. The powder
form of DHA oil was examined for its stability against oxidation as such
or in the form of a mixture with fish meal paste (eekamaboko''). The
powdery tridocosahexaenoin exhibited marked resistance against autoxidation
for a long period, during which the peroxide value remained constant at
a marginal level. Above all, POV for the powdery tridocosahexaenoin with
-CD remained virtually unchanged for 20 days without using any antioxidant
during storage at 4C, and seemed to be effective for fortifying fish
meal products.
Key words: tridocosahexaenoin; powdery encapsulation; X-ray diffractogram;
cyclodextrin
-30-
Note
Molecular Cloning and Characterization of a cDNA Encoding Putative Phospholipid
Hydroperoxide Glutathione Peroxidase from Spinach
Manabu Sugimoto, Satoshi Furui, and Yukio Suzuki
Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, Okayama 710, Japan
Received Jaunary 24, 1997
@A cDNA encoding spinach putative phospholipid hydroperoxide glutathione
peroxidase (PHGPX) was cloned and sequenced. The cDNA included an open
reading frame that encoded a polypeptide of 171 amino acid residues. The
deduced amino acid sequence showed about 77 and 50% similarity to plant
putative PHGPXs and mammalian PHGPXs, respectively. PCR products with the
same size as that of the spinach putative PHGPX were obtained from maize,
soybeans, and Arabidopsis, suggesting the expression of putative PHGPX
genes in other plants.
Key words: phospholipid hydroperoxide glutathione peroxidase;
Spinacia oleracea L.; gene cloning; nucleotide sequence
-31-
Note
Subsite Affinities of -Glucosidases from Spinach Seeds
Satoshi Furui, Manabu Sugimoto, and Yukio Suzuki
Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, Okayama 710, Japan
Received February 14, 1997
@The rate parameters for maltooligosaccharides of two -glucosidases
from spinach seeds were examined and subsite affinities were evaluated.
The subsite affinities for subsites 1, 2, 3, 4, 5, 6, and 7 in the active
site of -glucosidase A and B were 1.73, 2.91, 1.10, 0.25, 0.94, 0.03,
and -0.01 kcal/mol, and 0.33, 4.94, 0.26, 0.14, -0.24, -0.52, and 0.14
kcal/mol, respectively. The six and four subsites exist in -glucosidase
A and B, respectively, which maltooligosaccharides or soluble starch could
be bound.
Key words: -glucosidase; spinach seeds;
multiple molecular forms; subsite affinity
-32-
Note
Gibberellin Transport from the Cotyledon to Plumule in the Flowering of
Pharbitis nil
Listyani Wijayanti, Masatomo Kobayashi,*, Shozo Fujioka, and Akira Sakurai
The Institute of Physical and Chemical Research (RIKEN), Wako-shi, Saitama 351-01, Japan
Received February 12, 1997
@Uniconazole applied to cotyledons inhibited the flowering response in
Pharbitis nil. The application of gibberellin A1 (GA1) to cotyledons overcame
the effect of uniconazole and restored the flowering response. The transport
of GA1 from cotyledons to plumules was confirmed by GC-MS analyses. The
results indicate that GAs transported from cotyledons are concerned with
regulating the endogenous level of GAs in plumules.
Key words: flowering; gibberellin transport; Pharbitis nil
-33-
Note
O-Methylation of 2,6-Dimethoxy-4-methylphenol by Aspergillus glaucus and
Their Possible Contribution to Katsuobushi Flavor
Hajime Yamauchi and Mikiharu Doi
Marutomo Co., Ltd., 1696 Kominato, Iyo, Ehime 799-31, Japan
Received February 12, 1997
@Eleven strains of Aspergillus species, isolated from Katsuobushi (dried
bonito), were grown in a liquid medium containing 2,6-dimethoxy-4-methylphenol,
to examine the possibility of production of 1,2,3-trimethoxy-5-methylbenzene,
which has a musty odor, during the molding process in Katsuobushi production.
@In the liquid medium, 2,6-dimethoxy-4-methylphenol was O-methylated by
4 strains of A. glaucus.
Key words: Katsuobushi; Aspergillus species; O-methylation of phenols;
musty odor; molding
-34-
Note
Identification and NH2-terminal Amino Acid Sequences of DnaK and GroEL
Homologues in Moderate Eubacterial Halophiles
Masao Tokunaga, Kenzo Matsuoka, and Hiroko Tokunaga
Laboratory of Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890, Japan
Received February 12, 1997
@We have identified 2 DnaK and 3 GroEL homologues from moderately halophilic
Acinetobacter, Pseudomonas, and Planococcus species by partial purification
using an ATP-agarose column and by the analysis and similarity search of
these NH2-terminal amino acid sequences. Although these bacteria required
1 to 2 M NaCl for growth, these DnaK and GroEL homologues did not require
high salt to bind to the ATP column, thus suggesting that these chaperones
did not require high salts for their biochemically activities.
Key words: DnaK; GroEL; moderate eubacterial halophiles; NH2-terminal amino
acid sequence
-35-
Note
Ester Synthesis by NAD+-dependent Dehydrogenation of Hemiacetal: Production
of Methyl Formate by Cells of Methylotrophic Yeasts
Agung Primanto Murdanoto, Yasuyoshi Sakai, Langkah Sembiring, Yoshiki Tani, and Nobuo Kato
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa Sakyo-ku, Kyoto 606-01, Japan
Received February 14, 1997
@A water-soluble ester, methyl formate, was detected as a metabolite in
the culture medium of methylotrophic yeasts. Methyl formate synthase, which
catalyses NAD+-dependent dehydrogenation of the hemiacetal adduct of methanol
and formaldehyde, catalyses the ester synthesis. The enzyme activity was
induced on a methanol medium and was increased further by the addition
of formaldehyde. In the reaction system using intact cells of Pichia methanolica
AKU 4262, 135 mM (8.1 g/liter) methyl formate was produced from 2 M methanol.
This is a new biological process for ester synthesis that couples spontaneous
formation of hemiacetal and alcohol dehydrogenase.
Key words: formaldehyde oxidation; methanol; methyl formate; methyl formate
synthase; Pichia methanolica
-36-
Note
Highly Stereoselective Synthesis of (E)-Substituted Allylsilanes via the
Still-Wittig Rearrangement
Kazushige Fujii, Osamu Hara, and Youji Sakagami
Department of Applied Biological Sciences, Nagoya University, Chikusa-ku, Nagoya 464-01, Japan
Received March 6, 1997
@(E)- and (Z)-Vinylsilanes for the Still-Wittig rearrangement were easily
prepared from propargyl alcohol 4. The Still-Wittig rearrangement of (Z)-vinylsilane
1d afforded (E)-allylsilane 3d stereoselectively. The stereoselectivity
of (E)-vinylsilane, however, was unexpectedly low.
Key words: Still-Wittig rearrangement; (Z)-vinylsilane;
(E)-allylsilane
-37-
Note
4-O-Caffeoylshikimic and 4-O-(p-Coumaroyl)shikimic Acids from the Dwarf
Tree Fern, Dicksonia antarctica
Tamio Saito, Hisakazu Yamane,* Noboru Murofushi,** Nobutaka Takahashi, and Bernard O. Phinney***
The Institute of Physical and Chemical Research, Hirosawa 2-1, Wako-shi,
Saitama 351-01, Japan
* Biotechnology Research Center, The University of Tokyo, Yayoi 1-1-1,
Bunkyo-ku, Tokyo 113, Japan
** Department of Applied Biological Chemistry, The University of Tokyo,
Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan
*** Department of Biology, University of California, 405 Hilgard Ave.,
Los Angeles, CA 90024, U.S.A.
Received March 10, 1997
@Two derivatives of shikimic acid were isolated from croziers of the dwarf
tree fern, Dicksonia antarctica, and their structures were elucidated as
4-O-caffeoylshikimic acid and 4-O-(p-coumaroyl)shikimic acid on the basis
of mass spectrometric and NMR spectroscopic evidence.
Key words: caffeoyl ester; p-coumaroyl ester; Dicksonia antarctica; shikimic
acid; tree fern
-38-
Note
Lipid Peroxidation-derived Hepatotoxic Aldehyde, 4-Hydroxy-2-hexenal, in
Fish
Tadashi Sakai, Yoh-ichi Matsushita,* Kazuhiro Sugamoto,* and Koji Uchida**
Faculty of Agriculture, Miyazaki University, Miyazaki-shi, Miyazaki
889-21, Japan
* Faculty of Engineering, Miyazaki University, Miyazaki-shi, Miyazaki 889-21,
Japan
** Laboratory of Food and Biodynamics, Nagoya University School of Agriculture,
Nagoya 464-01, Japan
Received March 13, 1997
@Various samples of fish meat were examined for the formation of the hepatotoxic
aldehyde, 4-hydroxy-trans-2-hexenal (HHE). HHE was detected in all samples
analyzed at the concentration of 1.5-39.3 nmol/g. Yellowtail meat contained
more HHE than 4-hydroxy-trans-2-nonenal (HNE). The HHE and malonaldehyde
concentration increased during 13 days of storage at 0 C in the meat
of yellowtail. On the other hand, no HNE was detected during storage.
Key words: 4-hydroxy-2-hexenal; 4-hydroxy-2-nonenal; malonaldehyde; fish
meat
-39-
Rapid Paper
Evidence for the Periodically Alternating Microfibrillar Structure of Bacterial
Cellulose
Yoshinori Hori, 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 February 21, 1997
@The microfibrillar structure of bacterial cellulose was investigated.
Bacterial cellulose cultured in a jar fermentor was hydrolyzed in 1 N hydrochloric
acid for 2 h at 100C. The residual bacterial cellulose consisted of short
strands after hydrolysis. The lengths of 529 short strands on transmission
electron micrographs were measured with a ruler, and the length distribution
of the short strands was then evaluated by a computer-aided line shape
analysis. The length distribution was deconvoluted into five peaks with
Gaussian distribution, and the center positions of the peaks occurred at
regular intervals. This result demonstrates that the cellulose microfibrils
consisted of regularly repeating units of constant length (ca. 0.8 m)
and suggests that each unit consisted of an acid hydrolysis-susceptible
region (ca. 0.2 m) and an unsusceptible region (ca. 0.6 m).
Key words: bacterial cellulose; cellulose microfibril; acid hydrolysis;
periodicity
-40-
Short Communication
Unique Inhibition of Miltpain, a New Cysteine Proteinase from the Milt
of Chum Salmon, by o-Phenanthroline, Phenanthrenequinone, Phenazine, and
Acridine
Choko Kawabata and Eiji Ichishima*
Technical Research Center of T. Hasegawa Co., Ltd., Kariyado, 335 Nakahara-ku,
Kawasaki 211, Japan
* Laboratory of Molecular Enzymology, Department of Applied Biological
Chemistry, Faculty of Agriculture, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi,
Aoba-ku, Sendai 981, Japan
Received March 13, 1997
@Miltpain (EC 3.4.22.-) is a new cysteine proteinase that has been isolated
from the milt of chum salmon (Oncorhynchus keta), and is strongly inhibited
by cysteine proteinase inhibitors such as E-64, iodoacetamide, NEM, and
PCMB. This paper deals with a unique feature, that the enzyme becomes inhibited
by o-phenanthroline, phenanthrenequinone, phenanthrene, phenazine, and
acridine which have some structural resemblance to a hydrophobic planar
shape. Dixon plots showed that the inhibitions of the enzyme by o-phenanthroline
and phenanthrenequinone were noncompetitive.
Key words: cysteine proteinase; miltpain; o-phenanthroline; planar structure;
salmon proteinase
-41-
Short Communication
Pradimicin, a Mannose-binding Antibiotic, Induced Carbohydrate-mediated
Apoptosis in U937 Cells
Toshikazu Oki, Yoko Yamazaki, Tamotsu Furumai, and Yasuhiro Igarashi
Toyama Prefectural University, Biotechnology Research Center, 5180 Kurokawa, Kosugi, Toyama 939-03, Japan
Received April 7, 1997
@Pradimicin (PRM), a mannose-binding antifungal antibiotic, recognizes
a D-mannoside in the presence of calcium. We demonstrated that BMY-28864,
a semi-synthetic analog of PRM, induced apoptosis in U937 cells which had
been incubated with 1-deoxymannojirimycin (DMJ). Characteristic morphological
changes such as formation of apoptotic bodies and DNA fragmentation were
observed in apoptotic cells.
Key words: apoptosis; pradimicin; 1-deoxymannojirimycin
-42-
Short Communication
Structural Analyses of Xyloglucan Heptasaccharide by the Post-source Decay
Fragment Method Using Matrix-assisted Laser Desorption/Ionization Time-of-Flight
Mass Spectrometry
Tohru Yamagaki,*,** Yasushi Mitsuishi,* and Hiroshi Nakanishi*,
* National Institute of Bioscience and Human-Technology, AIST, 1-1 Higashi,
Tsukuba, Ibaraki 305, Japan
** Department of Biochemistry, Saitama University, Urawa, Saitama 338,
Japan
Received April 9, 1996
@In the post-source decay (PSD) fragment spectrum of a reduced xyloglucan
heptasaccharide (XXXGol) from tamarind seeds, eleven sodium-adduct fragment
ions and a precursor ion [M+Na]+ were clearly observed by using matrix-assisted
laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS).
Each fragment ion interval corresponded to the absence of unhydroxylose,
unhydroglucose, and glucitol residues, indicating that PSD fragmentation
cleavage in the sugar compound occurred only at glycosidic linkages close
to the oxygen atom of saccharide ring members, and not in inner sugar ring
bonds. The PSD fragment ions were classified into two series, one involving
the reducing end and the other involving the non-reducing end. Structural
information from both the reducing and non-reducing ends could therefore
be simultaneously obtained from the measurement of the positive ion mode.
Almost all the fragment ions from species larger than trisaccharide residues
could be detected in this PSD fragment experiment. Such fragmentation information
will enable the structural determination of xyloglucan oligosaccharides.
Key words: PSD fragment; MALDI-TOFMS; xyloglucan; oligosaccharide; fine
structural analysis
-43-
Short Communication
Synthesis and Biological Activities of Indolactone-V, the Lactone Analogue
of the Tumor Promoter (-)-Indolactam-V
Yu Nakagawa, Kazuhiro Irie, Yoshimasa Nakamura, Hajime Ohigashi, and Hideo Hayashi*
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto
University, Sakyo-ku, Kyoto 606, Japan
* Department of Applied Biochemistry, Faculty of Agriculture, University
of Osaka Prefecture, Sakai 593, Japan
Received May 1, 1997
@The tumor promoter (-)-indolactam-V (1) exists in two stable conformers
(twist and sofa) due to isomerization of the amide group. Indolactone-V
(2), the lactone analogue of 1, has been synthesized to investigate the
effects of the amide group on its conformation and biological activities.
Indolactone-V (2) existed only as the inactive sofa-like conformer, indicating
that the amide group of 1 plays a critical role in formation of the active
twist conformation.
Key words: conformation; (-)-indolactam-V; indolactone-V; protein kinase
C; teleocidin; tumor promoter