(Vol.66 No.1 2002)
Review
Moonlighting Functions of Polypeptide Elongation Factor 1: From Actin
Bundling to Zinc Finger Protein R1-Associated Nuclear Localization
Shin-ichiro EJIRI p.1
Inhibitory Effect of Isoflavones on Peroxynitrite-mediated Low-density
Lipoprotein Oxidation
Hsi-Huai LAI and Gow-Chin YEN† p.22
New Antioxidative Phenolic Glycosides Isolated from Kokuto Non-centrifuged
Cane Sugar
Kensaku TAKARA,† Daigo MATSUI, Koji WADA, Toshio ICHIBA,＊ and Yoko NAKASONE
p.29
Effect of Carnosine and Related Compounds on the Inactivation
of Human
Cu,Zn-Superoxide Dismutase by Modification of Fructose and Glycolaldehyde
Hiroyuki UKEDA,† Yuko HASEGAWA, Yumi HARADA and Masayoshi SAWAMURA p.36
Cloning and Overexpression of Bacillus cereus Penicillin-binding
Protein
3 gene in Escherichia coli
Takahisa MIYAMOTO, Md. Abu SAYED, Ryo SASAHARA, Kouji SUKIMOTO, Akiko UMEZAKI,
Ken-ichi HONJOH, Masayaoshi IIO, Shoji HATANO＊ p.44
Leaf-closing Substance in Leucaena leucocephala
Yoshihiro SOHTOME, Takashi TOKUNAGA, Katsuhiro UEDA,＃ Shosuke YAMAMURA,†
and Minoru UEDA† p.51
Purification and Properties of Membrane-bound D-Sorbitol Dehydrogenase
from Gluconobacter suboxydans IFO 3255＊
Teruhide SUGISAWA† and Tatsuo HOSHINO p.57
Polyphenol Increases in Safflower and Cucumber Seedlings Exposed
to Strong Visible Light with Limited Water
Satoe YAGINUMA,1 Takuo SHIRAISHI,1 Hiroaki OHYA,1,2 and Kiharu IGARASHI3 p.65
Kinetic Expression for the Oxidation of Linoleic
and Arachidonic Acid Esters in Their Mixed System
Eiichiro ISHIDO,1 Shuji ADACHI,1,† Yasumasa MINEMOTO,2 and Ryuichi MATSUNO1
p.73
Diverse Bacteria Related to the Bacteroides Subgroup of the
CFB Phylum
within the Gut Symbiotic Communities of Various Termites
Moriya OHKUMA,1,2,† Satoko NODA,1 Yuichi HONGOH,2 and Toshiaki KUDO1,2 p.78
Effects of GroESL Coexpression on the Folding of Nicotinoprotein
Formaldehyde Dismutase from Pseudomonas putida F61
Hideshi YANASE,† Keishi MORIYA, Norihiko MUKAI, Yasushi KAWATA,
Kenji OKAMOTO, and Nobuo KATO＊ p.85
Effects of Dietary Protein of Proso Millet on Liver Injury
Induced
by D-galactosamine in Rats
Naoyuki NISHIZAWA,1,† Daiki SATO,1 Yoshiaki ITO,1 Takashi NAGASAWA,1
Yasuko HATAKEYAMA,2 Myeong-Rak CHOI,3 You-Young CHOI,3 and Yi Min WEI4 p.92
A Novel Protein Toxin from the Deadly Box Jellyfish
(Sea Wasp, Habu-kurage) Chiropsalmus quadrigatus
Hiroshi NAGAI,＊,1,2 Kyoko TAKUWA-KURODA,＊ Masahiro NAKAO,†
Naomasa OSHIRO,ffl Setsuko IWANAGA,ffl and Terumi NAKAJIMA＊ p.97
On the Interaction of Ribosomal Protein L5 with 5S rRNA
Kenta IWASAKI,1 Shingo KIKUKAWA,1 Shunsuke KAWAMURA,2 Yoshiaki KOUZUMA,1
Isao TANAKA,3 and Makoto KIMURA1,† p.103
Effects of Amino Acid Alterations on the Transglycosylation
Reaction
of Endoglucanase I from Trichoderma viride HK-75
Il KWON, Keisuke EKINO, Takuji OKA, Masatoshi GOTO, and Kensuke FURUKAWA† p.110
Effects of Protein Deprivation on α1(I) and α1(III) Collagen
and Its Degrading System in Rat Skin
Yuichi OISHI,<0167> Zhengwei FU, Yoshinari OHNUKI, Hisanori KATO,† and
Tadashi NOGUCHI<0167> p.117
Substrate Specificity at the P1′ Site of Escherichia coli
OmpT under Denaturing Conditions
Kazuaki OKUNO,1,2,＊ Masayuki YABUTA,1 Kouji KAWANISHI,1 Kazuhiro OHSUYE,1
Toshihiko OOI,2 and Shinichi KINOSHITA2 p.127
3-(4-Methyl-3-pentenyl)-2(5H)-furanone, α,α-Acariolide
and 4-(4-Methyl-3-pentenyl)-2(5H)-furanone, α,β-Acariolide:
New Monoterpene Lactones from the Astigmatid Mites,
Schwiebea araujoae and Rhizoglyphus sp. (Astigmata: Acaridae)†
Hirokazu TARUI, Naoki MORI, Ritsuo NISHIDA, Kimiko OKABE＊ and Yasumasa KUWAHARA††
p.135
Differential Effect of Walnut Oil and Safflower Oil on the
Serum Cholesterol
Level and Lesion Area in the Aortic Root of Apolipoprotein E-deficient Mice
Masako IWAMOTO, Misaki KONO, Daisuke KAWAMOTO, Hiroko TOMOYORI,
Masao SATO, and Katsumi IMAIZUMI† p.141
The Primary Structure of Cassowary (Casuarius casuarius)
Goose Type Lysozyme
Sompong THAMMASIRIRAK, Takao TORIKATA, Kazutoshi TAKAMI,1
Koichi MURATA,2 and Tomohiro ARAKI† p.147
Amino Acid Sequence and Carbohydrate-binding Analysis
of the N-acetyl-D-galactosamine-specific C-Type Lectin, CEL-I,
from the Holothuroidea, Cucumaria echinata
Tomomitsu HATAKEYAMA,1,† Noriaki MATSUO,1 Kouhei SHIBA,1 Shoichi NISHINOHARA,2
Nobuyuki YAMASAKI,2 Hajime SUGAWARA,3 and Haruhiko AOYAGI1 p.157
Note
Purification and Some Properties of a Keratinolytic Enzyme
from an Alkaliphilic Nocardiopsis sp. TOA-1
Shinji MITSUIKI,1,3 Masashi SAKAI,1 Yasushi MORIYAMA,2 Masatoshi GOTO,3
and Kensuke FURUKAWA3 p.164
Note
Geraniol-inducible Glutathione S-Transferase in Cultured Soybean Cells
Yoshiyuki ASHIDA, Akihito MATSUSHIMA, Tomoko HIROTA,
Junko WATANABE, and Toshifumi HIRATA＊ p.168
Note
A Variant of Orpinomyces joyonii 1,3-1,4-β-Glucanase with Increased
Thermal Stability Obtained by Random Mutagenesis and Screening
Suk-Am KIM, Kuo-Joan CHENG, and Jin-Hao LIU† p.171
Note
Inhibition of Specific Degradation of 57-kDa Protein in Royal Jelly
during Storage by Ethylenediaminetetraacetic Acid
Masaki KAMAKURA† and Makoto FUKUSHIMA p.175
Note
Anti-inflammatory Compounds from the Bitter Mushroom, Sarcodon scabrosus
Mitsuru HIROTA,† Keiji MORIMURA, and Hisao SHIBATA p.179
Note
Inhibition of Translation and Progesterone-induced Maturation
of Xenopus Oocytes by Expressing the Amino-terminal Portion
of the Eukaryotic Translation Initiation Factor 4G
Motoaki WAKIYAMA and Kin-ichiro MIURA p.185
Note
Isolation of Yeast Kurtzmanomyces sp. I-11, Novel Producer
of Mannosylerythritol Lipid
Koji KAKUGAWA,1,2,† Masahiro TAMAI,3 Kunihiko IMAMURA,2,†† Keiko MIYAMOTO,3
Shozo MIYOSHI,2,††† Yuki MORINAGA,2,†††† Osamu SUZUKI,1 and Tokichi MIYAKAWA1
p.188
Note
Changes in N-Acetyl-β-D-Glucosaminidase Activity in the Urine
and Urinary Albumin Excretion in Magnesium Deficient Rats
Hiroshi MATSUZAKI,1 Junko OHDACHI,1 Masaaki FUCHIGAMI,1 Ritsuko MASUYAMA,2
Mariko UEHARA,2 Kahoru NAKAMURA,1 and Kazuharu SUZUKI2 p.192
Note
Uptake and physiological function of vitamin B12 in a photosynthetic
unicellular coccolithophorid alga, Pleurochrysis carterae
Emi MIYAMOTO,1 Fumio WATANABE,1,† Hiroyuki TAKENAKA,2 and Yoshihisa NAKANO3
p.195
Note
Use of Molecular-genetically Bred Coprinus cinereus Strains
for an Efficient Isolation of Cellulose from Rice Straw
Madoka KIKUCHI,† Kei-ichiro OGAWA, Takashi YAMAZAKI, Susumu KAJIWARA,
and Kazuo SHISHIDO p.199
Note
Effects of Heat and High-Pressure Treatments on Antigenicity of Beef Extract
Gi Dong HAN,a Masatomo MATSUNO,b Yoshihide IKEUCHI,c and Atsushi SUZUKId,†
p.202
Note
A Major Decomposition Product, Citrinin H2, from Citrinin
on Heating with Moisture
Mitsuru HIROTA,a,† Alka B. MENTA,b Keiko YONEYAMA,a and Naofumi KITABATAKEb
p.206
Note
TMSCl as a Mild and Effective Source of Acidic Catalysis in Fischer
Glycosidation and Use of Propargyl Glycoside for Anomeric Protection
Minoru IZUMI, Koichi FUKASE,† and Shoichi KUSUMOTO p.211
Note
Increased Antibody Production by Retinoids Is Related
to the Fusion Partner of Human Hybridomas
Yuichi INOUE and Sanetaka SHIRAHATA＊ p.215
Note
Decrease in Cytoplasmic pH-Homeostastatic Activity
of the Alkaliphile Bacillus Lentus C-125 by a Cell Wall Defect
Masahiro ITO1,＊ and Rikizo AONO1,† p.218
Preliminary Communication
Preparation and Catalytic Performance of Lipases Encapsulated
in Sol-Gel Materials
Katsuya KATO,† Yuefa GONG, Takao SAITO, and Yoshiyuki YOKOGAWA p.221
-1-
Review
Moonlighting Functions of Polypeptide Elongation Factor 1: From Actin
Bundling to Zinc Finger Protein R1-Associated Nuclear Localization
Shin-ichiro EJIRI
Cryobiosystem Research Center (CRC), Faculty of Agriculture, Iwate University, Morioka 020-8550, Japan
Eukaryotic polypeptide elongation factor EF-1 is not only a major translational factor, but also one of the most important multifunctional (moonlighting) proteins.
EF-1 consists of four different subunits collectively termed EF-1αββ′γ and EF-1αβγδ in plants and animals, respectively. EF-1α・GTP catalyzes the binding of aminoacyl-tRNA to the A-site of the ribosome. EF-1ββ′γ (EF-1β and EF-1β′), catalyzes GDP/GTP exchange on EF-1α・GDP to regenerate EF-1α・GTP. EF-1γ has recently been shown to have glutathione S-transferase activity.
EF-2 catalyzes the translocation of peptidyl-tRNA from the A-site to the P-site on the ribosome. Recently, molecular mimicry among tRNA, elongation factors, releasing factor (RF), and ribosome recycling factor (RRF) has been demonstrated and greatly improved our understanding of the mechanism of translation.
Moreover, eukaryotic elongation factors have been shown to be concerned or likely to be concerned in various important cellular processes or serious diseases, including translational control, signal transduction, cytoskeletal organization, apoptosis, adult atopic dermatitis, oncogenic transformation, nutrition, and nuclear processes such as RNA synthesis and mitosis.
This article aims to overview the recent advances in protein biosynthesis, concentrating on the moonlighting functions of EF-1.
Key words: protein biosynthesis; translation; elongation factor 1; cancer; apoptosis
-2-
Inhibitory Effect of Isoflavones on Peroxynitrite-mediated Low-density
Lipoprotein Oxidation
Hsi-Huai LAI and Gow-Chin YEN†
Department of Food Science, National Chung-Hsing University, 250 Kuokuang Road, Taichung 40227,
Taiwan, Republic of China
Received May 31, 2001; Accepted July 30, 2001
Peroxynitrite, a potent oxidant formed in vivo from the reaction of nitric oxide with superoxide, can mediate low-density liprotein (LDL) oxidation which is thought to increase the risk of atherosclerosis. This study investigates the inhibitory effect of the isoflavones, genistein and daidzein, together with their glycosidic forms, genistin and daidzin, on the peroxynitrite-mediated LDL oxidation and nitration of tyrosine. Genistein and daidzein were observed to dose-dependently inhibit peroxynitrite-mediated LDL oxidation, while their glucoside conjugates showed less activity. Moreover, all the isoflavones used in this study were found to be potent peroxynitrite scavengers, preventing the nitration of tyrosine. The ability of the isoflavones at 50 μM to decrease the tyrosine nitration induced by peroxynitrite (1 mM) was in the ratios of genistein (49％), daidzein (40％), daidzin (41％) and genistin (42％) when compared to the control (tyrosine incubated only with peroxynitrite). These results suggest that an intake of isoflavones could contribute to protecting against cardiovascular diseases and chronic inflammatory diseases.
Key words: isoflavone; peroxynitrite; LDL; tyrosine nitration
-3-
New Antioxidative Phenolic Glycosides Isolated from Kokuto Non-centrifuged
Cane Sugar
Kensaku TAKARA,† Daigo MATSUI, Koji WADA, Toshio ICHIBA,＊ and Yoko NAKASONE
Laboratory of Applied Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture,
University of the Ryukyus, Senbaru 1, Nishihara-cho, Okinawa 903-0213, Japan
＊Okinawa Industrial Technology Center, Suzaki 12-2, Gushikawa-shi, Okinawa, 904-2234, Japan
Received June 7, 2001; Accepted September 5, 2001
Nine compounds, 3-hydroxy-4,5-dimethoxyphenyl-β-D-glucopyranoside (1),β-D-fructfuranosyl-α-D-(6-vanilloyl)-glucopyranoside (2), β-D-fructfuranosyl-α-D-(6-syringyl)-glucopyranoside (3), 3-hydroxy-1-(4-hydroxy-
3-methoxyphenyl)-2-[4-(3-hydroxy-1-(E)-propenyl)-
2-methoxyphenoxy]propyl-β-D-glucopyranoside(4), 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-2-[4-(3-
hydroxy-1-(E)-propenyl)-2,6-dimethoxyphenoxy]
propyl-β-D-glucopyranoside (5), dehydrodiconiferyl
alcohol-9′-β-D-glucopyranoside (6), 4-[ethane-2-[3-(4-
hydroxy-3-methoxyphenyl)-2-propen]oxy]-2,6-dimethoxyphenyl-β-D-glucopyranoside (7), 4-[ethane-2-[3-(4-hydroxy-3-methoxyphenyl)-2-propen]oxy]-2-methoxyphenyl-β-D-glucopyranoside (8), and 3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-2-[4-(3-hydroxy-1-(E)-propenyl)-2,6-dimethoxyphenoxy]propyl-β-D-glucopyranoside (9), were isolated from Kokuto non-
centrifuged cane sugar. Their structures were elucidated by spectroscopic evidence, mainly based on the NMR technique. Among them, seven new glycosides were identified. The 2-deoxyribose oxidation method was used to measure their antioxidative activity. All of these compounds showed antioxidative activities.
Key words: cane sugar; Saccharum officinarum L.; Kokuto; antioxidant; glycoside
-4-
Effect of Carnosine and Related Compounds on the Inactivation of Human
Cu,Zn-Superoxide Dismutase by Modification of Fructose and Glycolaldehyde
Hiroyuki UKEDA,† Yuko HASEGAWA, Yumi HARADA and Masayoshi SAWAMURA
Department of Bioresources Science, Faculty of Agriculture, Kochi University, Monobe B-200,
Nankoku 783-8502, Japan
Received June 18, 2001; Accepted September 10, 2001
Glycolaldehyde, an intermediate of the Maillard reaction, and fructose, which is mainly derived from the polyol pathway, rapidly inactivate human Cu,Zn-superoxide dismutase (SOD) at the physiological concentration. We employed this inactivation with these carbonyl compounds as a model glycation reaction to investigate whether carnosine and its related compounds could protect the enzyme from inactivation. Of eight derivatives examined, histidine, Gly-His, carnosine and Ala-His inhibited the inactivation of the enzyme by fructose (p＜0.001), and Gly-His, Ala-His, anserine, carnosine, and homocarnosine exhibited a marked protective effect against the inactivation by glycolaldehyde (p＜0.001). The carnosine-related compounds that showed this highly protective effect against the inactivation by glycolaldehyde had high reactivity with glycolaldehyde and high scavenging activity toward the hydroxyl radical as common properties. On the other hand, the carnosine-related compounds that had a protective effect against the inactivation by fructose showed significant hydroxyl radical-scavenging ability. These results indicate that carnosine and such related compounds as Gly-His and Ala-His are effective anti-glycating agents for human Cu,Zn-SOD and that the effectiveness is based not only on high reactivity with carbonyl compounds but also on hydroxyl radical scavenging activity.
Key words: SOD; glycolaldehyde; fructose; glycation; carnosine
-5-
Cloning and Overexpression of Bacillus cereus Penicillin-binding Protein
3 gene in Escherichia coli
Takahisa MIYAMOTO, Md. Abu SAYED, Ryo SASAHARA, Kouji SUKIMOTO, Akiko UMEZAKI,
Ken-ichi HONJOH, Masayaoshi IIO, Shoji HATANO＊
Laboratory of Food Hygienic Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture,
Graduate School, Kyushu University, 6-10-1, Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan
＊Nishi-kyushu University, 4490-9, Ooazaozaki, kanzaki-cho, Kanzaki-gun, Saga 842-8585, Japan
Received June 25, 2001; Accepted September 11, 2001
The pbp3 gene encoding PBP3 of Bacillus cereus was cloned and sequenced. For this purpose, PBP3 was first purified from B. cereus ts-4, and N-terminal amino acid sequences of the peptides obtained from the protease digests of the protein were analyzed. The B. cereus ts-4 pbp3 gene consisted of an open reading frame of 1,986 bp encoding 662 amino acid residues with a calculated molecular mass of 73,044 Da. The active site-motifs SXXK, SXN, and KTG are present at the positions 393, 452, and 590, respectively, in the deduced amino acid sequence. The pbp3 structural gene was ligated into the pET17×b expression vector and pET-pbp3 was constructed. A protein was produced by the cells of E. coli carrying pET-pbp3. The produced protein migrated at about 75 kDa in SDS-polyacrylamide gel and strongly reacted with biotinylated ampicillin.
Key words: Bacillus cereus; PBP; sporulation; sequencing; overproduction
-6-
Leaf-closing Substance in Leucaena leucocephala
Yoshihiro SOHTOME, Takashi TOKUNAGA, Katsuhiro UEDA,＃ Shosuke YAMAMURA,†
and Minoru UEDA†
Laboratory of Natural Products Chemistry, Department of Chemistry, Faculty of Science and Technology,
Keio University, Hiyoshi, Yokohama 223-8522, Japan
＃Department of Chemistry, Biology and Marine Science, The University of Ryukyus, Nishihara-cho,
Okinawa 903-0213, Japan
Received July 2, 2001; Accepted September 19, 2001
Potassium (2R,3R)-2,3,4-trihydroxy-2-methylbutanoate
(1) was identified as a leaf-closing substance in the nyctinastic plant, Leucaena leucocephala. Compound 1 showed strong leaf-closing activity toward L. leucocephala and was not effective against other nyctinastic plants. The potassium ion was indispensable for the bioactivity of 1. Compound 1 gradually lost its bioactivity because of the exchange of the counter cation during isolation. A leaf-opening substance was also observed in the same plant.
Key words: nyctinasty; Leucaena leucocephala; leaf-closing substance; potassium (2R,3R)-2,3,4-trihydroxy-2-methylbutanoate
-7-
Purification and Properties of Membrane-bound D-Sorbitol Dehydrogenase
from Gluconobacter suboxydans IFO 3255＊
Teruhide SUGISAWA† and Tatsuo HOSHINO
Department of Applied Microbiology, Nippon Roche Research Center, 200 Kajiwara, Kamakura,
Kanagawa 247-8530, Japan
Received July 5, 2001; Accepted September 11, 2001
D-Sorbitol dehydrogenase was solubilized from the membrane fraction of Gluconobacter suboxydans IFO 3255 with Triton X-100 in the presence of D-sorbitol. Purification of the enzyme was done by fractionation with column chromatographies of DEAE-Cellulose, DEAE-Sepharose, hydroxylapatite, and Sephacryl HR300 in the presence of Triton X-100.
The molecular mass of the enzyme was 800 kDa, consisting of homologous subunits of 80 kDa. The optimum pH of the enzyme activity was 6.0, and the optimum temperature was 30°C.
Western blot analysis suggested the occurrence of the enzyme in all the Gluconobacter strains tested.
Key words: acetic acid bacteria; Gluconobacter strains; D-sorbitol dehydrogenase
-8-
Polyphenol Increases in Safflower and Cucumber Seedlings Exposed
to Strong Visible Light with Limited Water
Satoe YAGINUMA,1 Takuo SHIRAISHI,1 Hiroaki OHYA,1,2 and Kiharu IGARASHI3
1JST Regional Joint Research Project of Yamagata Prefecture, 2-2-1 Matsuei, Yamagata,
Yamagata 990-2473, Japan
2Institute for Life Support Technology, 2-2-1 Matsuei, Yamagata, Yamagata 990-2473, Japan
3Department of Bioresource Engineering, Faculty of Agriculture, Yamagata University, 1-23,
Wakaba-machi, Tsuruoka, Yamagata 997-8555, Japan
Received July 16, 2001; Accepted September 13, 2001
To assess effects of the environmental stress on polyphenol compounds (polyphenols) in plants, the polyphenol contents were investigated in the seedlings of safflower (Carthamus tinctrius L.) and cucumber (Cucumis sativus L.) grown under three types of growth conditions: control; light stress, irradiated with strong light in the visible wavelength range; and light/water stress, irradiated with strong visible light with a limited water supply. The total polyphenol contents and the amounts of the major polyphenols, especially luteolin 7-O-
glucoside in safflower cotyledons, and luteolin 7-O-glucoside and luteolin in safflower foliage leaves, increased in response to both stresses. The polyphenol increasing effect of light/water stress was clearly observed in safflower compared to cucumber, suggesting that plants that are resistant to these stresses can accumulate substantial amounts of polyphenols compared to the plants which respond weakly to the stresses.
Key words: safflower (Carthamus tinctrius L.); cucumber (Cucumis sativus L.); polyphenol; light stress; light/water stress
-9-
Kinetic Expression for the Oxidation of Linoleic
and Arachidonic Acid Esters in Their Mixed System
Eiichiro ISHIDO,1 Shuji ADACHI,1,† Yasumasa MINEMOTO,2 and Ryuichi MATSUNO1
1Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University,
Sakyo-ku, Kyoto 606-8502, Japan
2Department of Chemical and Biochemical Engineering, Toyama National College of Technology,
Hongo 13, Toyama 939-8630, Japan
Received July 18, 2001; Accepted September 6, 2001
Two polyunsaturated fatty acids (PUFAs) or their esters were mixed, and their oxidation processes were measured at 65°C and ca. 0％ relative humidity. Except when a PUFA ester was mixed with a free PUFA, the oxidation of the less-oxidative PUFA was promoted as its content in the mixture decreased, while the oxidation of the more-oxidative PUFA was delayed with a decrease in its content. A kinetic model is proposed whereby a PUFA acts as the diluent for another PUFA, and the oxidation rate of the PUFA is proportional to the product of the unoxidized PUFA concentration and the sum of the concentrations of the oxidized PUFA and the other oxidized PUFA. This model well expressed the oxidation processes of the PUFAs in their mixed system.
Key words: oxidation; polyunsaturated fatty acid; kinetic model
-10-
Diverse Bacteria Related to the Bacteroides Subgroup of the CFB Phylum
within the Gut Symbiotic Communities of Various Termites
Moriya OHKUMA,1,2,† Satoko NODA,1 Yuichi HONGOH,2 and Toshiaki KUD
O1,2 1Microbiology Laboratory, RIKEN (The Institute of Physical and Chemical Research) and
2International Cooperative Research Project, Japan Science and Technology Corporation
(JST-ICORP), Wako, Saitama 351-0198, Japan
Received July 19, 2001; Accepted September 14, 2001
Phylogenetically diverse clones of the partial 16S rDNA (ca. 850 bp) of bacteria belonging to the bacteroides subgroup of the cytophaga-flavobacter-bacteroides phylum were collected from the symbiotic microbial communities in the guts of six termite species without cultivation. Combined with the sequences reported previously, a total of thirty phylotypes of the subgroup were identified and classified into five phylogenetic clusters. One that was comprised of the phylotypes from a single termite species was related to the genus Rikenella. Two were clustered each with some cultured strains, genera of which have not been clearly defined yet. The remaining two clusters had no culturable representatives, suggesting the presence of yet-
uncultivated genera within the termite guts. From these sequence data, we designed a specific primer for the bacteroides subgroup, which was successful in the terminal-restriction fragment length polymorphism analysis to detect the phylotypes of the subgroup in the termite gut.
Key words: symbiosis; termite; the bacteroides subgroup; phylogeny; culture-independent approach
-11-
Effects of GroESL Coexpression on the Folding of Nicotinoprotein
Formaldehyde Dismutase from Pseudomonas putida F61
Hideshi YANASE,† Keishi MORIYA, Norihiko MUKAI, Yasushi KAWATA,
Kenji OKAMOTO, and Nobuo KATO＊
Department of Biotechnology, Faculty of Engineering, Tottori University, Tottori 680-8552, Japan
＊Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8224, Japan
Received July 23, 2001; Accepted August 31, 2001
The overexpression of fdm, which encodes the formaldehyde dismutase from Pseudomonas putida F61, resulted in the formation of inclusion bodies made up of aggregated enzyme, leaving little activity in the soluble fraction of the transformant cells. On the other hand, coexpression of groESL along with fdm facilitated in vivo solubilization of the enzyme protein in its active form. When coexpressed with groESL, formaldehyde dismutase purified from E. coli had the same crystalline form (i.e., a regular octahedron) as the native enzyme, and like the native enzyme, it bound 1 mol of NAD(H) and 2 mol of zinc in each subunit.
Key words: formaldehyde dismutase; GroESL; inclusion body; molecular chaperon; nicotinoprotein
-12-
Effects of Dietary Protein of Proso Millet on Liver Injury Induced
by D-galactosamine in Rats
Naoyuki NISHIZAWA,1,† Daiki SATO,1 Yoshiaki ITO,1 Takashi NAGASAWA,1
Yasuko HATAKEYAMA,2 Myeong-Rak CHOI,3 You-Young CHOI,3 and Yi Min WEI4
1Department of Agro-Bioscience, Faculty of Agriculture, Iwate University, Morioka, Iwate 020-8550, Japan
2Bellecenter Co. Ltd, Yahaba, Iwate 020-0891, Japan
3Division of Biotechnology and Chemical Engineering, Yosu National University, Yosu, Korea
4Northwest Science and Technology University of Agriculture and Forestry, Yangling, China
Received July 31, 2001; Accepted September 11, 2001
In this paper, we examined the effects of dietary protein from proso millet on liver injury induced by D-galactosamine or carbon tetrachloride in rats using serum enzyme activities as indices. D-galactosamine-
induced elevations of serum activities of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase were significantly suppressed by feeding the diet containing 20％ protein of proso millet for 14 days as compared with those of rats fed a 20％ casein diet, but not in the case of carbon tetrachloride. The results showed that proso millet protein is effective at lower dietary protein levels than that of dietary gluten reported previously. Therefore, the findings reported here may suggest that proso millet protein is considered to be another preventive food for liver injury.
Key words: proso millet; liver injury; D-galactosamine
-13-
A Novel Protein Toxin from the Deadly Box Jellyfish
(Sea Wasp, Habu-kurage) Chiropsalmus quadrigatus
Hiroshi NAGAI,＊,1,2 Kyoko TAKUWA-KURODA,＊ Masahiro NAKAO,†
Naomasa OSHIRO,ffl Setsuko IWANAGA,ffl and Terumi NAKAJIMA＊
＊Suntory Institute for Bioorganic Research, 1-1-1 Wakayamadai, Shimamoto, Mishima, Osaka 618-8503, Japan
†Institute for Biomedical Research, Suntory Ltd., 1-1-1 Wakayamadai, Shimamoto, Mishima,
Osaka 618-8503, Japan
fflOkinawa Prefectural Institute of Health and Environment, 2085 Ozato, Ozato-son, Okinawa 901-1202, Japan
Received July 31, 2001; Accepted August 30, 2001
The deadly box jellyfish (Sea Wasp, Habu-kurage in Japanese) Chiropsalmus quadrigatus Haeckel (Cubozoa) is distributed widely in the tropical Pacific region. In Japan, three fatal cases due to stings from this species have been reported officially. We successfully isolated C. quadrigatus toxin-A (CqTX-A, 44 kDa), a major proteinaceous toxin, for the first time, from the nematocysts of C. quadrigatus. CqTX-A showed lethal toxicity to crayfish when administered via intraperitoneal injection (LD50＝80 μg/kg) and hemolytic activity toward 0.8％ sheep red blood cells (ED50＝
160 ng/ml). Furthermore, we sequenced the cDNA encoding CqTX-A. The deduced amino acid sequence of CqTX-A (462 amino acids) showed 25.2％ and 21.6％ sequence similarity to Carybdea rastoni toxins (CrTXs) and Carybdea alata toxin-A (CrTX-A), respectively, which are Cubozoan jellyfish toxins.
Key words: jellyfish; Chiropsalmus quadrigatus; protein; hemolysis; toxin
-14-
On the Interaction of Ribosomal Protein L5 with 5S rRNA
Kenta IWASAKI,1 Shingo KIKUKAWA,1 Shunsuke KAWAMURA,2 Yoshiaki KOUZUMA,1
Isao TANAKA,3 and Makoto KIMURA1,†
1Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture,
Graduate School, Kyushu University, Fukuoka 812-8581, Japan
2Laboratory of Cell Biology, School of Agriculture, Kyushu Tokai University, Aso, Kumamoto 869-1404, Japan
3Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan
Received August 1, 2001; Accepted September 18, 2001
Ribosomal protein L5, a 5S rRNA binding protein in the large subunit, is composed of a five-stranded antiparallel β-sheet and four α-helices, and folds in a way that is topologically similar to the ribonucleprotein (RNP) domain [Nakashima et al., RNA 7, 692--701, 2001]. The crystal structure of ribosomal protein L5 (BstL5) from Bacillus stearothermophilus suggests that a concave surface formed by an anti-parallel β-sheet and long loop structures are strongly involved in 5S rRNA binding. To identify amino acid residues responsible for 5S rRNA binding, we made use of Ala-scanning mutagenesis of evolutionarily conserved amino acids occurred at β-strands and loop structures in BstL5. The mutation of Lys33 at the β1-strand caused a significant reduction in 5S rRNA binding. In addition, the Arg92, Phe122, and Glu134 mutations on the β2-strand, the α3-β4 loop, and the β4-β5 loop, respectively, resulted in a moderate decrease in the 5S rRNA binding affinity. In contrast, mutation of the conserved residue Pro65 at the β2-strand had little effect on the 5S rRNA binding activity. These results, taken together with previous results, identified Lys33, Asn37, Gln63, and Thr90 on the β-sheet structure, and Phe77 at the β2-β3 loop as critical residues for the 5S rRNA binding. The contribution of these amino acids to 5S rRNA binding was further quantitatively evaluated by surface plasmon resonance (SPR) analysis by the use of BIAcore. The results showed that the amino acids on the β-sheet structure are required to decrease the dissociation rate constant for the BstL5-5S rRNA complex, while those on the loops are to increase the association rate constant for the BstL5-5S rRNA interaction.
Key words: 5S rRNA binding protein; protein-RNA interaction; ribosomal protein L5; RNP domain; surface plasmon resonance
-15-
Effects of Amino Acid Alterations on the Transglycosylation Reaction
of Endoglucanase I from Trichoderma viride HK-75
Il KWON, Keisuke EKINO, Takuji OKA, Masatoshi GOTO, and Kensuke FURUKAWA†
Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University,
Fukuoka 812-8581, Japan
Received August 1, 2001; Accepted September 19, 2001
Endoglucanase I (EGI) from Trichoderma viride HK-75 catalyzes not only hydrolysis but also transglycosylation reactions of cellooligosaccharides. In order to characterize the important amino acid residues in transglycosylation of EGI, three Tyr, one Leu, and two Glu residues of EGI were replaced by Trp or Asp. The seven resulting EGI, except for L200W, had reduced activities toward carboxymethyl-cellulose compared to that of wild type EGI. The results from the mutations in the catalytic residues of E196 and E201 indicate that the space just around the catalytic residues is not directly related to the transglycosylation reactions of EGI. Analyses of the enzymes with mutations in the substrate-binding residues showed that Y146, Y170, and L200 of EGI are closely involved in the mode of transglycosylation and that several amino acid residues within the active site are involved in the transglycosylation reaction of EGI.
Key words: Trichoderma viride; cellulase; endoglucanase; transglycosylation; cellulose
-16-
Effects of Protein Deprivation on α1(I) and α1(III) Collagen
and Its Degrading System in Rat Skin
Yuichi OISHI,<0167> Zhengwei FU, Yoshinari OHNUKI, Hisanori KATO,† and Tadashi NOGUCHI<0167>
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences,
The University of Tokyo, Bunkyo-ku 113-8657, Tokyo, Japan
Received August 3, 2001; Accepted September 7, 2001
Protein malnutrition affects the status of dermal collagen, the major structural protein in the skin. However, the molecular mechanism underlying the alteration of collagen fibers in the skin by protein deficiency remains unknown. In the present study, the effect of dietary protein deprivation on collagen metabolism was studied by analyzing the status of the synthesis and degradation of collagen in the dorsal skin of rats. Feeding on a protein-free diet for 8 days caused a dramatic decrease in both types I and III tropocollagen with a concomitant decrease in their mRNA levels, with type III collagen being more severely affected. The active form of collagenase was significantly decreased by protein deprivation, whereas the latent form was not affected. The mRNA levels of collagenase and its inhibitors (TIMP-1 and 2) were also decreased by protein deprivation. These results suggest that both the synthesis and degradation of types I and III collagen were affected by protein deficiency.
Key words: protein deprivation; type I collagen; type III collagen; lysate RNase protection assay
-17-
Substrate Specificity at the P1′ Site of Escherichia coli
OmpT under Denaturing Conditions
Kazuaki OKUNO,1,2,＊ Masayuki YABUTA,1 Kouji KAWANISHI,1 Kazuhiro OHSUYE,1
Toshihiko OOI,2 and Shinichi KINOSHITA2
1Suntory Institute for Medicinal Research and Development, 2716-1 Kurakake, Akaiwa, Chiyoda-machi,
Ohra-gun, Gunma 370-0503, Japan
2Applied Biochemistry Laboratory, Division of Molecular Chemistry, Graduate School of Engineering,
Hokkaido University, N-13, W-8, Kita-Ku, Sapporo, Hokkaido 060-8628, Japan
Received August 6, 2001; Accepted September 17, 2001
Though OmpT has been reported to mainly cleave the peptide bond between consecutive basic amino acids, we identified more precise substrate specificity by using a series of modified substrates, termed PRX fusion proteins, consisting of 184 residues. The cleavage site of the substrate PRR was Arg140-Arg141 and the modified substrates PRX substituted all 19 natural amino acids at the P1′ site instead of Arg141. OmpT under denaturing conditions (in the presence of 4 M urea) cleaved not only between two consecutive basic amino acids but also at the carboxyl side of Arg140 except for the Arg140-Asp141, -Glu141, and -Pro141 pairs. In addition to Arg140 at the P1 site, similar results were obtained when Lys140 was substituted into the P1 site. In the absence of urea, an aspartic acid residue at the P1′ site was unfavorable for OmpT cleavage of synthetic decapeptides, the enzyme showed a preference for a dibasic site.
Key words: Escherichia coli; outer membrane protein OmpT endoprotease (EC 3.4.21.87); substrate specificity; denaturing conditions
-18-
3-(4-Methyl-3-pentenyl)-2(5H)-furanone, α,α-Acariolide
and 4-(4-Methyl-3-pentenyl)-2(5H)-furanone, α,β-Acariolide:
New Monoterpene Lactones from the Astigmatid Mites,
Schwiebea araujoae and Rhizoglyphus sp. (Astigmata: Acaridae)†
Hirokazu TARUI, Naoki MORI, Ritsuo NISHIDA, Kimiko OKABE＊ and Yasumasa KUWAHARA††
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku,
Kyoto 606-8502, Japan
＊Forest Insect Management Laboratory, Forestry and Forest Products Research Institute, Tsukuba Norin Kenkyu Danchi, P.O. Box 16, Ibaraki 305-8687, Japan
Received August 6, 2001; Accepted September 25, 2001
A new monoterpene lactone from the acarid mite, Schwiebea araujoae, was elucidated without its isolation by GC/FT-IR and GC/MS analyses to be 3-(4-methyl-3-pentenyl)-2(5H)-furanone (1) and tentatively named as α,α-acariolide. The structure of 1 was identified by its synthesis from α-bromo-γ-butyrolactone via 4 reaction steps. The synthesized compound gave the same GC/MS and GC/FT-IR spectra as those of the natural product.
The other monoterpene lactone was likewise elucidated from the unidentified Rhizoglyphus mite to be 4-(4-methyl-3-pentenyl)-2(5H)-furanone (2) and named as α,β-acariolide; it was also identified by its synthesis in 5 reaction steps from the same butyrolactone as the starting material. GC/MS and GC/FT-IR spectra of the preparation were identical to those of the natural product.
Key words: Schwiebea araujoae; monoterpene lactone; α,α-acariolide; 3-(4-methyl-3-pentenyl)-2(5H)-furanone; α,β-acariolide, 4-(4-methyl-3-pentenyl)-2(5H)-furanone
-19-
Differential Effect of Walnut Oil and Safflower Oil on the Serum Cholesterol
Level and Lesion Area in the Aortic Root of Apolipoprotein E-deficient Mice
Masako IWAMOTO, Misaki KONO, Daisuke KAWAMOTO, Hiroko TOMOYORI,
Masao SATO, and Katsumi IMAIZUMI†
Laboratory of Nutrition Chemistry, Graduate School of Agriculture, Kyushu University,
Fukuoka 812-8581, Japan
Recived August 8, 2001; Accepted September 17, 2001
Walnut oil (WO) is a good source of α-linolenic acid. We compared the effects of WO and high-linoleic safflower oil (HLSO) on the serum lipid level and atherosclerosis development in male and female apolipoprotein (apo) E-deficient mice. The WO diet resulted in a higher level of serum cholesterol than with HLSO. Female mice fed on the WO diet had a greater lesion area in the aortic root than did those on the HLSO diet. There was no diet-dependent difference in the level of cholesterol and its oxidation products in the abdominal and thoracic aorta. These results suggest that the unpleasant effects of the WO diet on apo E-deficient mice may be attributable to α-linolenic acid.
Key words: apolipoprotein E-deficient mice; atherosclerotic lesion; cholesterol; cholesterol oxidation product; linoleic acid
-20-
The Primary Structure of Cassowary (Casuarius casuarius)
Goose Type Lysozyme
Sompong THAMMASIRIRAK, Takao TORIKATA, Kazutoshi TAKAMI,1
Koichi MURATA,2 and Tomohiro ARAKI†
Department of Biochemistry, School of Agriculture, Kyushu Tokai University, Aso, Kumamoto,
869-1404, Japan
1Osaka Tennoji Zoo, 1-108, Chausuyama, Tennoji-ku, Osaka 543-0063, Japan
2Kobe Oji Zoo, 3-1, Oji-cho, Nada-ku, Kobe 657-0838, Japan
Received August 15, 2001; Accepted September 13, 2001
The complete amino acid sequence of cassowary (Casuarius casuarius) goose type lysozyme was analyzed by direct protein sequencing of peptides obtained by cleavage with trypsin, V8 protease, chymotrypsin, lysyl endopeptidase, and cyanogen bromide. The N-terminal residue of the enzyme was deduced to be a pyroglutamate group by analysis with a LC/MS/MS system equipped with the oMALDI ionization source, and then confirmed by a glutamate aminopeptidase enzyme. The blocked N-terminal is the first reported in this enzyme group. The positions of disulfide bonds in this enzyme were chemically identified as Cys4-Cys60 and Cys18-Cys29. Cassowary lysozyme was proved to consist of 185 amino acid residues and had a molecular mass of 20408 Da calculated from the amino acid sequence. The amino acid sequence of cassowary lysozyme compared to that of reported G-type lysozymes had identities of 90％, 83％, and 81％, for ostrich, goose, and black swan lysozymes, respectively. The amino acid substitutions at PyroGlu1, Glu19, Gly40, Asp82, Thr102, Thr156, and Asn167 were newly detected in this enzyme group. The substituted amino acids that might contribute to substrate binding were found at subsite B (Asn122Ser, Phe123Met). The amino acid sequences that formed three α-helices and three β-sheets were completely conserved. The disulfide bond locations and catalytic amino acid were also strictly conserved. The conservation of the three α-helices structures and the location of disulfide bonds were considered to be important for the formation of the hydrophobic core structure of the catalytic site and for maintaining a similar three-dimensional structure in this enzyme group.
Key words: cassowary; G-type lysozyme; amino acid sequence; lysozyme; disulfide bond
-21-
Amino Acid Sequence and Carbohydrate-binding Analysis
of the N-acetyl-D-galactosamine-specific C-Type Lectin, CEL-I,
from the Holothuroidea, Cucumaria echinata
Tomomitsu HATAKEYAMA,1,† Noriaki MATSUO,1 Kouhei SHIBA,1 Shoichi NISHINOHARA,2
Nobuyuki YAMASAKI,2 Hajime SUGAWARA,3 and Haruhiko AOYAGI1
1Department of Applied Chemistry, Faculty of Engineering, Nagasaki University, Bunkyo-machi 1-14,
Nagasaki 852-8521, Japan
2Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University, Hakozaki 6-10-1,
Fukuoka 812-8581, Japan
3Metabolic Function Research Group, RIKEN Plant Science Center, Hirosawa 2-1, Wako,
Saitama 351-0198, Japan
Received August 27, 2001; Accepted September 25, 2001
CEL-I is one of the Ca2＋-dependent lectins that has been isolated from the sea cucumber, Cucumaria echinata. This protein is composed of two identical subunits held by a single disulfide bond. The complete amino acid sequence of CEL-I was determined by sequencing the peptides produced by proteolytic fragmentation of S-pyridylethylated CEL-I. A subunit of CEL-I is composed of 140 amino acid residues. Two intrachain (Cys3-Cys14 and Cys31-Cys135) and one interchain (Cys36) disulfide bonds were also identified from an analysis of the cystine-containing peptides obtained from the intact protein. The similarity between the sequence of CEL-I and that of other C-type lectins was low, while the C-terminal region, including the putative Ca2＋ and carbohydrate-binding sites, was relatively well conserved. When the carbohydrate-binding activity was examined by a solid-phase microplate assay, CEL-I showed much higher affinity for N-acetyl-D-galactosamine than for other galactose-related carbohydrates. The association constant of CEL-I for p-nitrophenyl N-acetyl-β-D-galactosaminide (NP-GalNAc) was determined to be 2.3×104 M－1, and the maximum number of bound NP-GalNAc was estimated to be 1.6 by an equilibrium dialysis experiment.
Key words: C-type lectin; carbohydrate; amino acid sequence; peptide
-22-
Note
Purification and Some Properties of a Keratinolytic Enzyme
from an Alkaliphilic Nocardiopsis sp. TOA-1
Shinji MITSUIKI,1,3 Masashi SAKAI,1 Yasushi MORIYAMA,2 Masatoshi GOTO,3
and Kensuke FURUKAWA3
1Department of Industrial Chemistry, Kyushu Sangyo University, Fukuoka 813-8503, Japan
2Fundamental Research Center, TOTO Ltd., Kanagawa 253-8577, Japan
3Department of Bioscience and Biotechnology, Kyushu University, Fukuoka 812-8581, Japan
Received February 28, 2001; Accepted September 10, 2001
A novel alkaliphilic Nocardiopsis sp., strain TOA-1, was isolated from a tile-joint of a bathroom. Strain TOA-1 produced a variety of alkaline hydrolytic enzymes. An alkaline protease, designated NAPase, was purified and characterized. NAPase had a very high keratinolytic activity and high stability under acidic conditions.
Key words: alkaliphilic actinomycetes; Nocardiopsis sp.; alkaline protease; keratinolytic enzyme
-23-
Note
Geraniol-inducible Glutathione S-Transferase in Cultured Soybean Cells
Yoshiyuki ASHIDA, Akihito MATSUSHIMA, Tomoko HIROTA,
Junko WATANABE, and Toshifumi HIRATA＊
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University,
1-3-1 Kagamiyama, Higashi-Hiroshima 739-8526, Japan
Received May 29, 2001; Accepted September 14, 2001
When the cultured cells of Glycine max (soybean) were treated with 5 mM geraniol as a chemical stress, an mRNA level was elevated in a rapid but transient increase. The mRNA was cloned and sequenced, and found to correspond to the mRNA encoding glutathione S-transferase (GST). The GST mRNA level and GST activity were elevated to maxima at 4--6 h and 8 h, respectively, after treatment of the cultures with geraniol. These indicate that GST is one of the geraniol-responsive factors in soybean cells.
Key words: Glycine max; geraniol; glutathione S-transferase; chemical stress; defense reaction
-24-
Note
A Variant of Orpinomyces joyonii 1,3-1,4-β-Glucanase with Increased
Thermal Stability Obtained by Random Mutagenesis and Screening
Suk-Am KIM, Kuo-Joan CHENG, and Jin-Hao LIU†
Institute of BioAgricultural Sciences, Academia Sinica, Taipei, Taiwan 115, Republic of China
Received June 29, 2001; Accepted September 22, 2001
A thermostable variant of an Orpinomyces joyonii β-glucanase was identified by screening a mutant library constructed using error-prone PCR products. The mutant, designated 2011D, had one amino acid substitution (Val replaced Asp-70). 2011D showed similar catalytic efficiency to its wild-type enzyme, LicA. The temperature at which 50％ inactivation occurred after heat treatment for 10 min was increased by 14°C for 2011D, in comparison to those of wild-type enzyme.
Key words: β-glucanase; thermal stability; random mutagensis; DNA shuffling
-25-
Note
Inhibition of Specific Degradation of 57-kDa Protein in Royal Jelly
during Storage by Ethylenediaminetetraacetic Acid
Masaki KAMAKURA† and Makoto FUKUSHIMA
POLA R＆D Laboratories, POLA Corporation, 560 Kashio-cho, Totsuka-ku, Yokohama 244-0812, Japan
Received July 2, 2001; Accepted September 2, 2001
We have previously shown that 57-kDa protein in royal jelly (RJ) was specifically degraded in proportion to both storage temperature and storage period, and we suggested that it could be useful as a marker of freshness of RJ (Kamakura, M., Fukuda, T., Fukushima, M. and Yonekura, M., Biosci. Biotechnol. Biochem., 65, 277--284 (2001).). Here, we investigated the effects of various proteinase inhibitors on proteinase activity in RJ and on the specific degradation of 57-kDa protein during storage. Ethylenediaminetetraacetic acid (EDTA), but not other inhibitors, inhibited the proteinase activity in RJ, and dose-dependently suppressed storage-dependent degradation of 57-kDa protein. These results suggest that EDTA inhibits a specific proteinase activity in RJ, thereby suppressing the degradation of 57-kDa protein during storage at high temperature.
Key words: royal jelly; 57-kDa protein; metalloproteinase; ethylenediaminetetraacetic acid; storage-dependent degradation
-26-
Note
Anti-inflammatory Compounds from the Bitter Mushroom, Sarcodon scabrosus
Mitsuru HIROTA,† Keiji MORIMURA, and Hisao SHIBATA
Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University,
8304 Minami-minowa, Kami-ina, Nagano 399-4598, Japan
Received July 6, 2001; Accepted September 7, 2001
A bioassay-guided purification procedure from the methanol extract of Sarcodon scabrosus led to the isolation of several anti-inflammatory compounds: sarcodonin A (1) and G (2), and related compounds (3, 4 and 5). We named these related compounds neosarcodonin A (3), B (4) and C (5) and elucidated their structures on the basis of spectral data. Topical application of each of these compounds to mouse ears suppressed TPA-
induced inflammation. Neosarcodonin C (5) exhibited the highest activity and inhibited the TPA-induced edema on mouse ears by up to 87％ with a 200-μg application.
Key words: anti-inflammation; Sarcodon scabrosus; TPA; neosarcodonin
-27-
Note
Inhibition of Translation and Progesterone-induced Maturation
of Xenopus Oocytes by Expressing the Amino-terminal Portion
of the Eukaryotic Translation Initiation Factor 4G
Motoaki WAKIYAMA and Kin-ichiro MIURA
Institute for Biomolecular Science, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo 171-8588, Japan
Received July 12, 2001; Accepted September 12, 2001
The eukaryotic translation initiation factor 4G (eIF4G) plays a pivotal role in translation. EIF4G interacts with several other factors including eIF4E, which is a cap-binding protein, and the poly(A)-binding protein (PABP). In this work, we demonstrate that the expression of the amino-terminal one-third of eIF4G, which interacts with eIF4E and PABP, in Xenopus oocyte inhibits translation and progesterone-induced maturation.
Key words: eukaryotic translation initiation factor 4G; Xenopus; oocyte; maturation
-28-
Note
Isolation of Yeast Kurtzmanomyces sp. I-11, Novel Producer
of Mannosylerythritol Lipid
Koji KAKUGAWA,1,2,† Masahiro TAMAI,3 Kunihiko IMAMURA,2,†† Keiko MIYAMOTO,3
Shozo MIYOSHI,2,††† Yuki MORINAGA,2,†††† Osamu SUZUKI,1 and Tokichi MIYAKAWA1
1Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter,
Hiroshima University, 1-3-1, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8530, Japan
2Hiroshima Prefectural Institute of Industrial Science and Technology, 3-10-32, Kagamiyama,
Higashi-Hiroshima, Hiroshima 739-0046, Japan
3Hiroshima Prefectural Food Technology Research Center, 12-70, Hijiyama-honmachi, Minami-ku,
Hiroshima 732-0816, Japan
Received July 12, 2001; Accepted August 29, 2001
Yeast strains were screened for producers of glycolipid-type biosurfactants from soybean oil as a sole carbon source. The structure of the glycolipid (MEL-
I-11) produced by strain I-11 was analyzed. The hydrophilic sugar moiety was mannosylerythritol and the fatty acid components were C8:0 (36.4％), C12:0 (11.9％), and C14:2 (25.9％). The MEL-I-11 was identified as 6-O-acetyl-2,3-di-O-alkanoyl-β-D-
mannopyranosyl-(1→4)-O-meso-erythritol. The strain I-11 was identified as a Kurtzmanomyces species, a novel producer of mannosylerythritol lipid.
Key words: biosurfactant; mannosylerythritol lipid; Kurtzmanomyces sp. yeast
-29-
Note
Changes in N-Acetyl-β-D-Glucosaminidase Activity in the Urine
and Urinary Albumin Excretion in Magnesium Deficient Rats
Hiroshi MATSUZAKI,1 Junko OHDACHI,1 Masaaki FUCHIGAMI,1 Ritsuko MASUYAMA,2
Mariko UEHARA,2 Kahoru NAKAMURA,1 and Kazuharu SUZUKI2
1Department of Nutrition, Junior College of Tokyo University of Agriculture, 1-1-1 Sakuragaoka,
Setagaya-ku, Tokyo 156-8502, Japan
2Department of Nutritional Science, Faculty of Applied Bioscience, Tokyo University of Agriculture,
1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan
Received July 17, 2001; Accepted September 13, 2001
To discover the details of the effects of magnesium (Mg) deficiency on kidney function, the course of changes in N-acetyl-β-D-glucosaminidase (NAG) activity in the urine and in urinary albumin excretion were examined in rats fed a Mg-deficient diet. NAG activity in the urine and urinary albumin excretion in rats fed the Mg-deficient diet significantly increased from 7 d until the end of the feeding period. We suggest that Mg-deficient diet rapidly induces kidney function insufficiency.
Key words: magnesium deficiency; N-acetyl-β-D-glucosaminidase activity in urine; urinary albumin excretion; rats
-30-
Note
Uptake and physiological function of vitamin B12 in a photosynthetic
unicellular coccolithophorid alga, Pleurochrysis carterae
Emi MIYAMOTO,1 Fumio WATANABE,1,† Hiroyuki TAKENAKA,2 and Yoshihisa NAKANO3
1Department of Health Science, Kochi Women′s University, Kochi 780-8515, Japan
2MicroAlgae Corporation, Tokyo 104-0061, Japan
3Division of Applied Biochemistry, Graduate School of Agriculture and Biological Sciences,
Osaka Prefecture University, Sakai 599-8531, Japan
Received July 18, 2001; Accepted September 10, 2001
The photosynthetic coccolithophoid alga, Pleurochrysis (Hymenomonas) carterae, could take up and accumulate exogenous vitamin B12, most of which was converted into the coenzyme forms of vitamin B12. Two vitamin B12-dependent enzyme activities (methylmalonyl-CoA mutase, 2.6±0.4 nmol/min/mg protein and methionine synthase, 85.1±38.9 pmol/
min/mg protein) could be found in a cell homogenate of the vitamin B12-supplemented alga. Most of the methylmalonyl-CoA mutase activity and 19.2％ of the vitamin B12 accumulated by the algal cells were recovered in the macromolecular fractions with Mr of 150 kDa, although the remaining vitamin B12 was found only in free vitamin B12 fractions.
Key words: cobalamin; coccolithophorid; methylmalonyl-CoA mutase; Pleurochysis carterae; vitamin B12
-31-
Note
Use of Molecular-genetically Bred Coprinus cinereus Strains
for an Efficient Isolation of Cellulose from Rice Straw
Madoka KIKUCHI,† Kei-ichiro OGAWA, Takashi YAMAZAKI, Susumu KAJIWARA,
and Kazuo SHISHIDO
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology,
Nagatuta, Midori-ku, Yokohama 226-8501, Japan
Received July 23, 2001; Accepted September 17, 2001
Molecular-bred Coprinus cinereus monokaryotic strains with high lignin- and xylan-degrading activities were mixed-cultured at 27°C in the liquid medium containing 0.5％(w/v) cut rice straw and 0.025％ MnCl2. After 3 weeks, the culture supernatant was extensively treated with crude cellulase, showing the presence in it of 9.3％ of the total cellulose of rice straw. When rice straw treated with 0.1 N NaOH or cultured with Ganoderma applanatum were used, the recoveries of the cellulose increased up to 29％. The same experiments were done by using a non-bred control strain, showing the recoveries of the cellulose from the treated or cultured rice straw to be 8％.
Key words: basidiomycete mushroom; lignin degradation; molecular breeding; plant biomass use; xylan degradation
-32-
Note
Effects of Heat and High-Pressure Treatments on Antigenicity of Beef Extract
Gi Dong HAN,a Masatomo MATSUNO,b Yoshihide IKEUCHI,c and Atsushi SUZUKId,†
aDoctor′s Program in Functional Biology, Graduate School of Science and Technology,
University of Niigata, Niigata 950-2181, Japan
bDepartment of Pediatrics, Yoshida Hospital, Yoshida 959-9213, Japan
cDivision of Food Animal Science, Graduate School of Agricultural Sciences, Kyushu University,
Fukuoka 812-8581, Japan
dDepartment of Applied Biological Chemistry, Faculty of Agriculture, University of Niigata,
Niigata 950-2181, Japan
Received July 23, 2001; Accepted August 30, 2001
The sera of bovine gamma globulin (BGG) positive beef allergic patients were used in this study in order to investigate changes in IgE-specific binding activity with regard to beef extract altered by heat or high-pressure treatment. In inhibition-ELISA, the sample treated at 60°C did not show any significant changes in the antigenicity of BGG, but the sample treated at 100°C showed a decrease of the antigenicity. In the case of the treatment with heating at 100°C, heat-coagulation occurred in the beef extract. The resulting supernatant and precipitate of the sample by centrifugation were analyzed by immunoblotting. Only the fraction of precipitate showed a specific binding activity with the sera. Based on this result, it was speculated that the persistent antigenicity found even after the treatment at 100°C in inhibition-ELISA remained principally in the heat-coagulated fraction, which indicated the importance of the method of handling the heat-coagulation in heat treatment. High-pressure treatments (200 MPa--
600 MPa) of beef extract did not show any significant changes in the binding with the sera.
Key words: antigenicity/allergenicity; heat treatment; high-pressure treatment; bovine gamma globulin; beef extract
-33-
Note
A Major Decomposition Product, Citrinin H2, from Citrinin
on Heating with Moisture
Mitsuru HIROTA,a,† Alka B. MENTA,b Keiko YONEYAMA,a and Naofumi KITABATAKEb
aDepartment of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University,
8304 Minami-minowa, Kami-ina, Nagano 399-4598, Japan
bResearch Institute for Food Science, Kyoto University, Uji, Kyoto 611-0011, Japan
Received July 27, 2001; Accepted September 7, 2001
Citrinin is one of the mycotoxins produced by Penicillium citrinum. We examined the decomposition products after heating citrinin in water at 140°C and isolated a major product, citrinin H2 (3-(3,5-dihydroxy-2-methylphenyl)-2-formyloxy-butane). Citrinin H2 did not show significant cytotoxicity to HeLa cells up to a concentration of 200 μg/ml (％ cytotoxicity: 39％) in 63 h of incubation, but citrinin showed severe toxicity at a concentration of 25 μg/ml (％ cytotoxicity: 73％). HPLC analysis of citrinin after heating under various conditions indicates that citrinin H2 is mainly yielded from citrinin.
Key words: citrinin; detoxified product; HeLa cells; cytotoxicity
-34-
Note
TMSCl as a Mild and Effective Source of Acidic Catalysis in Fischer
Glycosidation and Use of Propargyl Glycoside for Anomeric Protection
Minoru IZUMI, Koichi FUKASE,† and Shoichi KUSUMOTO
Department of Chemistry, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan
Received August 6, 2001; Accepted September 25, 2001
Practical Fischer glycosidation was effected at room temperature or 60°C by using 5 to 10 equiv. of TMSCl. The anomeric propargyl group formed by this method was found to be a versatile new protecting group, being stable in neat TFA but readily cleaved by treatment with Co2(CO)8 and TFA in CH2Cl2 via the formation of an alkyne--Co complex.
Key words: Fischer glycosidation; protective group; TMSCl; acid catalyst
-35-
Note
Increased Antibody Production by Retinoids Is Related
to the Fusion Partner of Human Hybridomas
Yuichi INOUE and Sanetaka SHIRAHATA＊
Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University,
1-21-24 Korimoto, Kagoshima 890-0065, Japan
＊Graduate School of Genetic Resources Technology, Kyushu University, 6-10-1 Hakozaki, Higashi-ku,
Fukuoka 812-8581, Japan
Received August 10, 2001; Accepted September 14, 2001
An increase of human monoclonal antibody production caused by retinyl acetate and retinoic acid was influenced by the fusion partner rather than the original B lymphocyte used for the human hybridoma generation. Retinoid response of human hybridomas may be at least related to retinoid X receptor-alpha gene expression, which seemed to originate from their fusion partner.
Key words: antibody production; fusion partner; human monoclonal antibody; hybridoma; retinoids
-36-
Note
Decrease in Cytoplasmic pH-Homeostastatic Activity
of the Alkaliphile Bacillus Lentus C-125 by a Cell Wall Defect
Masahiro ITO1,＊ and Rikizo AONO1,†
1Department of Biological Information, Graduate School of Bioscience and Biotechnology,
Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-8501, Japan
Received August 10, 2001; Accepted September 18, 2001
Cytoplasmic pH homeostatic activities of cell wall-defective derivatives of the alkaliphile Bacillus lentus C-125 were assessed using a pH-sensitive fluorescent probe, BCECF. It was shown that the acidic cell wall components took part in maintenance of the cytoplasmic pH neutrality at alkaline pH.
Key words: alkaliphile; alkaliphilic Bacillus lentus; cell wall; cytoplasmic pH; pH homeostasis
-37-
Preliminary Communication
Preparation and Catalytic Performance of Lipases Encapsulated
in Sol-Gel Materials
Katsuya KATO,† Yuefa GONG, Takao SAITO, and Yoshiyuki YOKOGAWA
Bio-functional Ceramics Group, Ceramics Research Institute, National Institute of Advanced Industrial Science
and Technology (AIST), 2266-98 Anagahora, Simoshidami, Moriyama-ku, Nagoya 463-8560, Japan
Received August 28, 2001; Accepted October 16, 2001
Three kinds of lipases (from Candida antarctica, Pseudomonas cepacia, and Pseudomonas fluorescens) were encapsulated in inorganic matrices by the sol-gel method in order to synthesize chiral compounds by kinetic resolution. Sol-gel lipases prepared with vinyltriethoxysilane had higher hydrolysis activity for 2-octyl acetate than those with other silane precursors: tetramethoxysilane, methyltrimethoxysilane, and propyltrimethoxysilane.
Key words: lipase; enantioselectivity; sol-gel; encapsulated; vinyltriethoxysilane<QL>