(Vol.62 No.12 1998)
Review
Relationship between Chemical Structures and Biological Activities of Triterpenoid
Saponins from Soybean
Yumiko YOSHIKI, Sigemitsu KUDOU,＊ and Kazuyoshi OKUBO
Extraction of Water-soluble Soybean Polysaccharides under Acidic
Conditions
Hitoshi FURUTA, Taro TAKAHASHI, Jyunko TOBE, Ryosuke KIWATA, and Hirokazu MAEDA＊
Hydroxyl Radical Generation Depending on O2 or H2O by a Photocatalyzed
Reaction in an Aqueous Suspension of Titanium Dioxide
Hitoshi SHIBATA,・ Yasushi OGURA, Yoshihiro SAWA, and The lateYasuhisa KONO
Purification and Characterization of tert-Butyl Ester-hydrolyzing
Lipase from Burkholderia sp. YY62
Soo-Hwan YEO, Takuya NIHIRA,・ and Yasuhiro YAMADA
Molecular Cloning and Sequencing of Two Phospho-β-galactosidase
I and II
Genes of Lactobacillus gasseri JCM1031 Isolated from Human Intestine
Tadao SAITO,＊,・ Masakatsu SUZUKI,＊ Kei KONNO,＊ Haruki KITAZAWA,＊
Yasushi KAWAI,＊ Takatoshi ITOH,＊ and Yoshiyuki KAMIO＊＊
Improved Bioassay Method for Plant Transformation Inhibitors
Hiroshi KANZAKI,・ Toshihiko KAGEMORI, Satomi ASANO, and Kazuyoshi KAWAZU
Arginine Enhances Induction of T Helper 1 and T Helper 2 Cytokine
Synthesis by Peyer′s Patch αβ T Cells and Antigen-Specific Mucosal Immune Response
Toshiya KOBAYASHI,＃,・,＊ Masafumi YAMAMOTO,＃,・ Takachika HIROI,・ Jerry MCGHEE,＃
Yasuyoshi TAKESHITA,・ and Hiroshi KIYONO＃,・
Purification and Partial Identification of Bacteriocin ISK-1,
a New Lantibiotic
Produced by Pediococcus sp. ISK-1
Hirokazu KIMURA, Hiromi MATSUSAKI, Toshihiro SASHIHARA, Kenji SONOMOTO,
and Ayaaki ISHIZAKI＊
Transformation of the Edible Basidiomycete Lentinus edodes by
Restriction
Enzyme-Mediated Integration of Plasmid DNA
Toshitsugu SATO, Kaori YAEGASHI, Shizuko ISHII, Tatsuya HIRANO,
Susumu KAJIWARA,＊ Kazuo SHISHIDO,＊ and Hitoshi ENEI
Apoptosis Induced by Niacin-related Compounds in HL-60 Cells
Shin OGATA, Masayo TAKEUCHI, Katsuzumi OKUMURA, and Hiroshi TAGUCHI・
Alanine Dehydrogenase from Enterobacter aerogenes: Purification,
Characterization, and Primary Structure
Emran Kabir CHOWDHURY,1 Tetsuya SAITOH,1 Shinji NAGATA,1 Makoto ASHIUCHI,2
and Haruo MISONO1,2,・
Isolation of the creA Gene from the Cellulolytic Fungus Humicola
grisea and
Analysis of CreA Binding Sites Upstream from the Cellulase Genes
Shou TAKASHIMA,・ Akira NAKAMURA, Makoto HIDAKA, Haruhiko MASAKI,
and Takeshi UOZUMI・・
NapA Na＋/H＋ Antiporter as a Sodium Extrusion System Supplementary
to the Vacuolar Na＋-ATPase in Enterococcus hirae
Miyuki KAWANO,＊ Kazuei IGARASHI,＊ Marc SOLIOZ,＊＊ and Yoshimi KAKINUMA＊,・
Cloning, Sequencing, High Expression, and Crystallization
of the Thermophile Thermus aquaticus Glycerol Kinase
Hua-Shan HUANG, Kiyoshi ITO, Chang-Hong YIN, Tsutomu KABASHIMA,
and Tadashi YOSHIMOTO・
Synthesis of Optically Active 1,4-Thiazane-3-carboxylic
Acid via Optical Resolution by Preferential Crystallization
of (RS)-2-Amino-3-[(2-chloroethyl)sulfanyl]propanoic
Acid Hydrochloride
Tadashi SHIRAIWA,・ Kohya TADOKORO, Haruyuki TANAKA, Keiichiro NANBA,
Noriyoshi YOKONO, Katsuyoshi SHIBAZAKI, Motoki KUBO,
and Hidemoto KUROKAWA
A Novel Glycerol Kinase from Flavobacterium meningosepticum:
Characterization, Gene Cloning and Primary Structure
Shin-ichi SAKASEGAWA, Issei YOSHIOKA, Shinzi KOGA, Mamoru TAKAHASHI,
Kunio MATSUMOTO,＊ Hideo MISAKI, and Toshihisa OHSHIMA＊＊
Substrate Specificity of 2-Deoxy-scyllo-inosose Synthase, the
Starter Enzyme
for 2-Deoxystreptamine Biosynthesis, toward Deoxyglucose-6-phosphates
and Proposed Mechanism・
Noriaki IWASE, Fumitaka KUDO, Noriaki YAMAUCHI, and Katsumi KAKINUMA＊
Cloning of the Gene for Inorganic Pyrophosphatase from a Thermoacidophilic
Archaeon, Sulfolobus sp. Strain 7, and Overproduction of the Enzyme
by Coexpression of tRNA for Arginine Rare Codon
Takayoshi WAKAGI,1,・ Tairo OSHIMA,2 Hiromi IMAMURA,1 and Hiroshi MATSUZAWA1
Note
Transglycosylation of Thiamin by Fungal β-N-Acetylhexosaminidases
Vladimir K・REN,1 Zdenka HU・NKOVA,1 Petr HALADA,1 and Yukio SUZUKI2
Note
Nutritional Effects of a D-Methionine-containing Solution
on AH109A Hepatoma-bearing Rats
Taizo SASAMURA, Akihiko MATSUDA, and Yukifumi KOKUBA
Note
Analysis of the Oligosaccharide Units of Xyloglucans by Digestion
with Isoprimeverose-producing Oligoxyloglucan Hydrolase Followed
by Anion-exchange Chromatography
Teruko KONISHI,・ Yasushi MITSUISHI,＊ and Yoji KATO・・
Note
Cloning of Genomic DNA of Rhizopus niveus Lipase and Expression
in the Yeast Saccharomyces cerevisiae
Mitsutaka KOHNO, Makoto ENATSU, and Wataru KUGIMIYA
Note
Simple Synthesis of Both Enantiomers of the C7-C12 Segment of Epothilones
Shinji TANIMORI,＊ Koichi TANIMOTO, and Mitsunori KIRIHATA
Note
Purification of Pumpkin Glutathione S-Transferase Species Specifically Present
in Cultured Cells Treated by Excessive Concentration
of 2,4-Dichlorophenoxyacetic Acid but Absent in Normal Plants
Masayuki FUJITA,＊ Yasuhiko ADACHI, and Nobuo SAKATO
Note
Lipase-catalyzed Asymmetric Synthesis of Enantiomerically Pure
(2S,4aS,8S)-4a,8-Dimethyl-2,3,4,4a,5,6,7,8-octahydro-2-naphthalenol
Akira TANAKA,1,・ Takahito TOKUYAMA,2 Akiko SAITO,1 and Takayuki ORITANI2
Note
Gassericin A; an Uncommon Cyclic Bacteriocin Produced by Lactobacillus gasseri
LA39 Linked at N- and C-Terminal Ends
Yasushi KAWAI,・ Tadao SAITO, Haruki KITAZAWA, and Takatoshi ITOH
Note
Promotion by a Peptidyl Growth Factor, Phytosulfokine, of Chlorophyll
Formation in Etiolated Cotyledon of Cucumber
Seiyei YAMAKAWA,1 Yoshikatsu MATSUBAYASHI,2 Youji SAKAGAMI,2
Hiroshi KAMADA,1 and Shinobu SATOH1
Note
Inhibition by a Capsaicin Antagonist (Capsazepine) of Capsaicin-induced
Swimming Capacity Increase in Mice
Kyung-Mi KIM, Teruo KAWADA, Kengo ISHIHARA, Kazuo INOUE,
and Tohru FUSHIKI・
Note
Cloning and Nucleotide Sequence of the Gene Encoding a Serine Proteinase
Inhibitor Named Marinostatin from a Marine Bacterium,
Alteromonas sp. Strain B-10-31
Katsushiro MIYAMOTO,1 Hiroshi TSUJIBO,1,・ Yumi HIKITA,1 Kazumi TANAKA,1
Satomi MIYAMOTO,1
Mariko HISHIMOTO,1 Chiaki IMADA,2 Kaeko KAMEI,3 Saburo HARA,3 and Yoshihiko
INAMORI1
Note
New Antibacterial Diterpenoids from the Sarcodon scabrosus Fungus＊
Hisao SHIBATA,・ Atsushi IRIE, and Yasufumi MORITA
Note
Thermal Isomerization of All-trans-Lutein in a Benzene Solution
Achmad SUBAGIO,1 Naofumi MORITA,1,・ and Shigeo SAWADA2
Note
Compilation of mRNA Sequences Surrounding the AUG Translation
Initiation Codon in the Green Alga Chlamydomonas reinhardtii
Kazunori IKEDA and Hitoshi MIYASAKA＊,・
Note
Novel Cyclohexene Compound from Lasiodiplodia theobromae IFO 31059
Hideyuki MATSUURA, Naomi OBARA, Noriko CHISAKA, Akitami ICHIHARA,
and Teruhiko YOSHIHARA・
Note
Synthesis of a Novel Phosphate Ester of a Vitamin E Derivative
and Its Antioxidative Activity
Sayuri MIYAMOTO,・・,＊ Takuro KOGA,＊＊ and Junji TERAO・,＊＊＊
Note
Prevention of Initial Supercooling in Progressive Freeze-concentration
Ling LIU,・ Tomoyuki FUJII, Kiro HAYAKAWA,＊ and Osato MIYAWAKI・・
Preliminary Communication
Structural Analysis of Xyloglucan Oligosaccharides by the Post-source
Decay Fragmentation Method of MALDI-TOF Mass Spectrometry:
Influence of the Degree of Substitution by Branched Galactose, Xylose,
and Fucose on the Fragment Ion Intensities
Tohru YAMAGAKI,a,b Yasushi MITSUISHI,a and Hiroshi NAKANISHIa＊
Preliminary Communication
Identification of a Structural Gene Encoding a Metallothionein-like Domain
that Includes a Putative Regulator Protein for Streptomyces
Protease Gene Expression
Seiichi TAGUCHI,＊ Takahiro OGAWA, Takeshi ENDO, and Haruo MOMOSE
Preliminary Communication
Biosynthesis of striatol in cultured cells of liverwort, Ptycanthus striatus
Kenji KATOH,1 Kanae YAMAMOTO,2 Masateru YAMAMOTO,2 Makoto HASHIMOTO,2
-1-
Review
Relationship between Chemical Structures and Biological Activities
of Triterpenoid Saponins from Soybean
Yumiko YOSHIKI, Sigemitsu KUDOU,＊ and Kazuyoshi OKUBO
Department of Environmental Bioremediation, Graduate School of Agricultural Chemistry,
Tohoku University, 1-1 Tsutsumidori Amamiyamachi, Aoba-ku, Sendai, Miyagi 981-8555, Japan
＊Kanesa Company, Ltd., Tamagawa, Aomori 038-0000, Japan
Saponins can be found in more than one hundred plant families and in some marine animals. However, chemical investigation of soybean (Glycine max L. Merrill) saponins has begun only as recently as the 1970s. Here we focus on the chemical structure, the content, and biological activity of soybean saponins in current studies. Especially, we focus on 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) conjugated saponins and define the chemical structure of this as a natural precursor of group B and E saponins.
soybean; saponin; triterpenoid; Leguminosae; soyasaponin
-2-
Extraction of Water-soluble Soybean Polysaccharides under Acidic Conditions
Hitoshi FURUTA, Taro TAKAHASHI, Jyunko TOBE, Ryosuke KIWATA, and Hirokazu MAEDA＊
Novelty Research Institute, Tsukuba R＆D Center, Fuji Oil Co. Ltd., 4-3 Kinunodai, Yawara-mura, Tsukuba-gun, Ibaraki 300-2497, Japan
＊Protein Development Dept. II, Hannan R＆D Center, Fuji Oil Co. Ltd., 1 Sumiyoshi-cho, Osaka 598-0061, Japan
Received June 1, 1998; Accepted August 13, 1998
We investigated the yield, gelation and sugar composition of water-soluble polysaccharides that had been extracted from soybean cotyledons under acidic conditions. SSPS were easily extracted at several pHs. The highest yield was about twice that reported by Morita et al., after extraction in boiling water, or by Kawamura et al., after extraction in hot water (90°C) with the stepwise addition of ammonium oxalate and 0.5％ NaOH. Our results show that the average molecular weight was relatively high for both SSPS extracted in the weakly acidic pH range of 4--6 at 100--120°C and in the strongly acidic pH range of 2--3 at 40--80°C. The water-soluble polysaccharide solutions which were extracted under the former conditions remained fluid, but those which were extracted under the latter conditions gelled. However, the molecular weight distribution and sugar composition were similar for SSPS isolated by using both sets of conditions. Furthermore, water-soluble soybean polysaccharides were found to consist of rhamnogalacturonans, which could endure hydrolysis, and arabinogalactans, which were easily hydrolyzed under acidic conditions.
soybean; polysaccharide; extraction; acidity
-3-
Hydroxyl Radical Generation Depending on O2 or H2O by a Photocatalyzed
Reaction in an Aqueous Suspension of Titanium Dioxide
Hitoshi SHIBATA,・ Yasushi OGURA, Yoshihiro SAWA, and The lateYasuhisa KONO
Department of Life Science and Biotechnology, Faculty of Life and Environmental Science,
Shimane University, Matsue, Shimane 690-8504, Japan
Received June 8, 1998; Accepted September 9, 1998
Photocatalytic production of the electron (e－) and positive hole (h＋) in an aqueous suspension of TiO2 (anatase form) under illumination by near-UV light (295--390 nm) generated the superoxide (O－2) and hydroxyl radical (・OH), which both proceeded linearly with reaction time, while H2O2 accumulated non-linearly. Under anaerobic conditions (introduced Ar gas), the yields of three active species of oxygen were decreased to 10--20％ of those detected in the air-saturated reaction. The electron spin resonance (ESR) signal characteristics of ・OH were obtained when a spin trap of 5,5-dimthyl-1-pyrroline-N-oxide (DMPO) was included in the illuminating mixture. The intensity of the ESR signal was increased by Cu/Zn superoxide dismutase, and decreased under anaerobic conditions, amounting to only 20％ of the intensity detected in the aerobic reaction. The addition of H2O2 to the reaction mixture resulted in about an 8-fold increase of ・OH production in the anaerobic reaction, but only about 1.5-fold in the aerobic reaction, indicating that e－ generated by the photocatalytic reaction reduced H2O2 to produce ・OH plus OH－. On the other hand, D2O lowered the yield of ・OH generation to 18％ under air and 40％ under Ar conditions, indicating the oxidation of H2O by h＋. The addition of Fe(III)-EDTA as an electron acceptor effectively increased ・OH generation, 2.3-fold in the aerobic reaction and 8.4-fold in the anaerobic reaction, the yield in the latter exceeding that in the air-saturated reaction.
hydroxyl radical; photocatalyst; titanium dioxide; near-UV light; water oxidation
-4-
Purification and Characterization of tert-Butyl Ester-hydrolyzing Lipase
from Burkholderia sp. YY62
Soo-Hwan YEO, Takuya NIHIRA,・ and Yasuhiro YAMADA
Department of Biotechnology, Graduate School of Engineering, Osaka University,
2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
Received June 10, 1998; Accepted August 14, 1998
An intracellular novel lipase which can hydrolyze t-butyl octanoate (TBO) was purified to homogeneity from crude cell-free extracts of Burkholderia (formerly Pseudomonas) sp. YY62 with 9％ overall yield. Seventy-four-fold purification was achieved by ammonium-sulfate precipitation, three consecutive open-column chromatographies (DEAE anion-exchange, Sepharose CL-6B gel-filtration, and the second DEAE anion-exchange columns), and two HPLCs (TSK G2000SWXL gel-filtration and phenyl 5PW hydrophobic interaction columns). Enzymes hydrolyzing p-nitrophenyl acetate were separated into two peaks (peak I and II) on the hydrophobic HPLC, and only peak II was found to have TBO-hydrolyzing activity. The peak preparation showed a single band of 40 kDa on SDS-PAGE and a molecular mass of 39 kDa on gel-filtration under non-denatured conditions, indicating the monomeric nature of the TBO-hydrolyzing lipase. The lipase showed maximum activity at pH 7.0 and 28°C. The N-terminal 15 amino acid residues were determined as Met-Asp-Phe-Tyr-Asp-Ala-Asn-Glu-Thr-Arg-His-Pro-Glu-Gln-Arg, which showed no homology to known proteins, suggesting that the purified enzyme may belong to a novel class of hydrolase.
Burkholderia sp.; t-butyl octanoate; esterase; lipase
-5-
Molecular Cloning and Sequencing of Two Phospho-β-galactosidase I and II
Genes of Lactobacillus gasseri JCM1031 Isolated from Human Intestine
Tadao SAITO,＊,・ Masakatsu SUZUKI,＊ Kei KONNO,＊ Haruki KITAZAWA,＊
Yasushi KAWAI,＊ Takatoshi ITOH,＊ and Yoshiyuki KAMIO＊＊
＊Laboratory of Animal Products Chemistry, ＊＊Laboratory of Applied Microbiology, Graduate School
of Agricultural Science, Tohoku University, Tsutsumidori Amamiyamachi, Aoba-ku, Sendai 981-8555, Japan
Received June 10, 1998; Accepted August 13, 1998
Lactobacillus (Lb.) gasseri JCM1031, which is classified into the B1 subgroup of the Lb. acidophilus group of lactic acid bacteria, characteristically produces two different phospho-β-galactosidases (P-β-gal) I and II in the same cytosol as reported in our previous papers [Biosci. Biotech. Biochem., 60, 139--141, 708--710 (1996)]. To clarify the functional and genetic properties of the two enzymes, the structural genes of P-β-gal I and II were cloned and sequenced. The structural gene of P-β-gal I had 1,446 bp, encoding a polypeptide of 482 amino acid residues. The structural gene of P-β-gal II had 1,473 bp, encoding a polypeptide of 491 amino acid residues. The deduced relative molecular masses of 55,188 and 56,243 agreed well with the previous value obtained from the purified P-β-gal I and II protein, respectively. Multiple alignment of the protein sequence of P-β-gal I and II with those of P-β-gals from 5 microorganisms had 30--35％ identity on the amino acid level, but those with phospho-β-glucosidases from 5 microorganisms had the relatively high identity of about 50％. Considering that this strain grows on lactose medium and shows no β-galactosidase activity, and that purified P-β-gal I and II can obviously hydrolyze o-nitrophenyl-β-D-galactopyranoside 6-phosphate (substrate), and also the conservation of a cysteine residue in the molecule, the P-β-gal I and II were each confirmed as a novel P-β-gal enzyme.
Lactobacillus gasseri; phospho-β-galactosidase; molecular cloning; structural gene
-6-
Improved Bioassay Method for Plant Transformation Inhibitors
Hiroshi KANZAKI,・ Toshihiko KAGEMORI, Satomi ASANO, and Kazuyoshi KAWAZU
Laboratory of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Okayama 700-8530, Japan
Received June 11, 1998; Accepted September 4, 1998
A convenient and quantitative bioassay method for evaluating the efficiency of plant transformation by Agrobacterium tumefaciens is important to search plant transformation inhibitors, possible biochemical probes for study on its mechanism. Our previously reported method, in which the plant transformation had been detected by the expression of β-glucuronidase in transformed plants, was improved. The difference between the previous and the improved methods is the use of suspension-cultured cells of Ageratum conyzoides as the host plant instead of Nicotiana tabacum BY-2; this alteration of the host enabled us to measure the β-glucuronidase activity in plant cells not only fluorometrically but also colorimetrically. The enzyme activity expressed in the cells of A. conyzoides was nearly 100 times higher than that of N. tabacum BY-2, and was enough for detection by colorimetric measurement. The method, therefore, is useful for a convenient determination of inhibitory activity against plant transformation.
Agrobacterium tumefaciens; GUS assay; Ageratum conyzoides; colorimetric measurement; fluorometric measurement
-7-
Arginine Enhances Induction of T Helper 1 and T Helper 2 Cytokine Synthesis
by Peyer′s Patch αβ T Cells and Antigen-Specific Mucosal Immune Response
Toshiya KOBAYASHI,＃,・,＊ Masafumi YAMAMOTO,＃,・ Takachika HIROI,・ Jerry MCGHEE,＃
Yasuyoshi TAKESHITA,・ and Hiroshi KIYONO＃,・
＃Departments of Oral Biology and Microbiology, Immunobiology Vaccine Center, The University of Alabama at Birmingham, Medical Center, Birmingham, Alabama 35294, U.S.A.
・Life Science Research Institute, Snow Brand Milk Products Co. Ltd., Tochigi 329-0512, Japan
・Department of Mucosal Immunology, Research Institute for Microbial Diseases, Osaka University,
Suita, Osaka 565-0871, Japan
Received June 17, 1998; Accepted August 5, 1998
The effects of arginine on cell proliferation and subsequent T helper (Th) 1 and Th 2 cytokine synthesis by murine Peyer′s patch (PP) Th cells in vitro and the influence of arginine on the induction of antigen-specific mucosal and systemic immune responses in vivo were examined. When the PP T cells were stimulated with the anti-αβ T cell receptor (TCR) antibody in the presence of different concentrations of arginine, a higher proliferative response was observed in the culture with an optimal concentration of arginine compared with that with a minimum amount of this amino acid. The concentration of cytokines in the supernatant, the number of cytokine-producing cells and the cytokine-specific mRNA expression of PP T cells were also increased in a dose-dependent fashion. Furthermore, when mice fed on an arginine-supplemented liquid diet were orally immunized with tetanus toxoid plus cholera toxin as a mucosal adjuvant, a higher level of antigen-specific fecal IgA was observed when compared with the response in mice fed on an arginine-free diet. Taken together, these results suggest that arginine enhanced antigen-specific mucosal immune response resulting from the supporting activation of cell proliferation and subsequent cytokine synthesis of PP Th cells.
arginine; Peyer′s patch T cell; cytokine synthesis; antigen-specific immune response
-8-
Purification and Partial Identification of Bacteriocin ISK-1, a New Lantibiotic
Produced by Pediococcus sp. ISK-1
Hirokazu KIMURA, Hiromi MATSUSAKI, Toshihiro SASHIHARA, Kenji SONOMOTO,
and Ayaaki ISHIZAKI＊
Laboratory of Microbial Technology, Department of Food Science and Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan
Received June 24, 1998; Accepted August 25, 1998
Bacteriocin ISK-1 is a proteinaceous inhibitory substance produced by Pediococcus sp. ISK-1 isolated from well-aged Nukadoko. Bacteriocin ISK-1 was purified by acid treatment, ammonium sulfate precipitation, cation-exchange chromatography, and reversed-phase HPLC from the culture supernatant of Pediococcus sp. ISK-1. Purification of bacteriocin ISK-1 resulted in a 30-fold increase in the specific activity and the recovery was 17％. Molecular mass of bacteriocin ISK-1 measured by fast atom bombardment-mass spectrometry was 2,960. The amino acid composition analysis of bacteriocin ISK-1 showed that it contained unusual amino acids such as lanthionine and/or 3-methyllanthionine, which is a characteristic of lantibiotics. The N-terminal amino acid sequence analysis indicated the first seven N-terminal amino acid residues as NH2-K-K-K-S-G-V-I. The primary sequence showed significant similarity to the lantibiotics lacticin 481 from Lactococcus lactis and variacin from Micrococcus varians, which suggests that bacteriocin ISK-1 is a novel lantibiotic belonging to a lacticin-481 type.
Pediococcus; bacteriocin; lantibiotic; Nukadoko
-9-
Transformation of the Edible Basidiomycete Lentinus edodes by Restriction
Enzyme-Mediated Integration of Plasmid DNA
Toshitsugu SATO, Kaori YAEGASHI, Shizuko ISHII, Tatsuya HIRANO,
Susumu KAJIWARA,＊ Kazuo SHISHIDO,＊ and Hitoshi ENEI
Iwate Biotechnology Research Center, 22-174-4 Narita, Kitakami, Iwate 024-0003, Japan
＊Department of Life Science, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology,
Nagatsuta, Yokohama-shi, Kanagawa 226-8501, Japan
Received July 2, 1998; Acceped August 26, 1998
We have used the restriction enzyme-mediated DNA integration (REMI) method to establish a transformation system in Lentinus edodes using the recombinant plasmid pLC1-hph, which contains the L. edodes transcriptional signals and an Escherichia coli hygromycin B phosphotransferase gene. Protoplasts of L. edodes were treated by the PEG transformation mixture containing 50 units of SalI, which cleaves pLC1-hph at a single site, yielding about 15 transformants per 2.5 μg of DNA. The conventional PEG transformation without SalI, however, yielded only 1.5 transformants per 25 μg of DNA. The optimal amount of SalI for increased transformation was 50 units. In the case of transformation with SphI, which cleaves the plasmid at one site, the optimal amount of the enzyme was 2.5 units. Southern blot analysis of the SphI-derived transformants suggested that 50％ of the plasmid integrations were REMI events.
Lentinus edodes; REMI; transformation; hygromycin B
-10-
Apoptosis Induced by Niacin-related Compounds in HL-60 Cells
Shin OGATA, Masayo TAKEUCHI, Katsuzumi OKUMURA, and Hiroshi TAGUCHI・
Laboratory of Biological Chemistry, Faculty of Bioresouces, Mie University, Tsu, Mie 514-8507, Japan
Received July 6, 1998; Accepted August 14, 1998
We have investigated whether niacin-related compounds act as inducers of apoptosis in HL-60 cells. In this study, we found that picolinic acid, dipicolinic acid, and isonicotinamide strongly induce apoptosis. After treatments with these compounds, apoptosis started within 4 h and was induced in about 50％ of the cells within 8 h. These compounds induced apoptosis at 5--10 mM, but did not at 1 mM. An ICE-like protease inhibitor (Z-Asp-CH2-DCB) completely blocked the apoptosis, but a caspase-1 inhibitor (Ac-YVAD-CHO) and a caspase-3 inhibitor (Ac-DEVD-CHO) did not block the apoptosis, suggesting that other caspases have the critical roles in the execution process of apoptosis induced by niacin-related compounds.
niacin; NAD; poly(ADP-ribose) polymerase; apoptosis; HL-60
-11-
Alanine Dehydrogenase from Enterobacter aerogenes: Purification,
Characterization, and Primary Structure
Emran Kabir CHOWDHURY,1 Tetsuya SAITOH,1 Shinji NAGATA,1 Makoto ASHIUCHI,2
and Haruo MISONO1,2,・
Laboratory of Applied Microbiology, Department of Bioresources Science,1 and Research Institute
of Molecular Genetics,2 Kochi University, Nankoku, Kochi 783-8502, Japan
Received July 6, 1998; Accepted August 17, 1998
Alanine dehydrogenase [EC 1. 4. 1. 1] was purified to homogeneity from a crude extract of Enterobacter aerogenes ICR 0220. The enzyme had a molecular mass of about 245 kDa and consisted of six identical subunits. The enzyme showed maximal activity at about pH 10.9 for the deamination of L-alanine and at about pH 8.7 for the amination of pyruvate. The enzyme required NAD＋ as a coenzyme. Analogs of NAD＋, deamino-NAD＋ and nicotinamide guanine dinucleotide served as coenzymes. Initial-velocity and product inhibition studies suggested that the deamination of L-alanine proceeded through a sequential ordered binary-ternary mechanism. NAD＋ bound first to the enzyme, followed by L-alanine, and the products were released in the order of ammonia, pyruvate, and NADH. The Km were 0.47 mM for L-alanine, 0.16 mM for NAD＋, 0.22 mM for pyruvate, 0.067 mM for NADH, and 66.7 mM for ammonia. The Km for L-alanine was the smallest in the alanine dehydrogenases studied so far. The enzyme gene was cloned into Escherichia coli JM109 cells and the nucleotides were sequenced. The deduced amino acid sequence was very similar to that of the alanine dehydrogenase from Bacillus subtilis. However, the Enterobacter enzyme has no cysteine residue. In this respect, the Enterobacter enzyme is different from other alanine dehydrogenases.
alanine dehydrogenase; Enterobacter aerogenes; kinetic mechanism; primary structure; nucleotide sequence
-12-
Isolation of the creA Gene from the Cellulolytic Fungus Humicola grisea and
Analysis of CreA Binding Sites Upstream from the Cellulase Genes
Shou TAKASHIMA,・ Akira NAKAMURA, Makoto HIDAKA, Haruhiko MASAKI,
and Takeshi UOZUMI・・
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo,
Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan
Received July 7, 1998; Accepted August 28, 1998
A carbon catabolite repressor gene, creA, was isolated from the cellulolytic fungus Humicola grisea by using a portion of the Trichoderma reesei cre1 gene as a probe. The deduced amino acid sequence predicts a zinc finger protein of 419 amino acids in length, and its zinc finger regions show high similarity with those of Aspergillus CreAs, T. reesei Cre1, and Saccharomyces cerevisiae MIG1. Northern blot analysis showed that the H. grisea creA gene was highly transcribed when the mycelia were grown on glucose-containing media, but the transcription of the H. grisea endoglucanase 1 gene (egl1) and the exoglucanase 1 gene (exo1) were repressed under these conditions. Results of binding assays with the maltose-binding protein::CreA(1--166) fusion protein and the egl1 and the exo1 upstream regions showed that some 6-bp sites having an identical or similar sequence to the consensus sequence for CreA binding were protected from DNase I digestion.
carbon catabolite repression; cellulase; creA; Humicola grisea; zinc finger protein
-13-
NapA Na＋/H＋ Antiporter as a Sodium Extrusion System Supplementary
to the Vacuolar Na＋-ATPase in Enterococcus hirae
Miyuki KAWANO,＊ Kazuei IGARASHI,＊ Marc SOLIOZ,＊＊ and Yoshimi KAKINUMA＊,・
＊Faculty of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan
＊＊Department of Clinical Pharmacology, University of Berne, Murtenstrasse 35, 3010 Berne, Switzerland
Received July 9, 1998; Accepted July 30, 1998
Enterococcus hirae has two sodium extrusion systems: the NapA Na＋/H＋ antiporter and the vacuolar Na＋-ATPase. We found that a NapA mutant, WD4, which is deficient in Na＋/H＋ antiporter activity, grew well in the pH range of 6 to 10 up to 200 mM sodium. This was due to active, potential-independent sodium extrusion by the Na＋-ATPase, which was induced under these conditions. The NapA Na＋/H＋ antiporter is thus not a prerequisite for growth of E. hirae in the presence of sodium, but plays a supplementary role in sodium extrusion at acidic pH.
NapA; Na＋/H＋ antiporter; sodium extrusion; vacuolar Na＋-ATPase; Enterococcus hirae
-14-
Cloning, Sequencing, High Expression, and Crystallization
of the Thermophile Thermus aquaticus Glycerol Kinase
Hua-Shan HUANG, Kiyoshi ITO, Chang-Hong YIN, Tsutomu KABASHIMA,
and Tadashi YOSHIMOTO・
School of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-machi,
Nagasaki, Nagasaki 852, Japan
Received July 9, 1998; Accepted August 31, 1998
Glycerol kinase (EC 2.7.1.30) is a key enzyme of glycerol uptake and metabolism in bacteria. Using PCR, we amplified and cloned a glycerol kinase gene, glpK, from Thermus aquaticus. The complete gene has 1488 base pairs, coding for a protein of 496 amino acids with a predicted molecular weight of 54,814. The amino acid sequence deduced from T. aquaticus glpK was found to have identities of 97 and 81％, respectively, with those of Thermus flavus and Bacillus subtilis glpK genes. After overproduction in Escherichia coli, the expressed enzyme was easily purified to homogeneity by DEAE-Toyopearl chromatography. The purified enzyme has been crystallized by the hanging drop vapor diffusion method at 22°C. Comparison of the amino acid sequence with that of the B. subtilis enzyme showed that Ser and Lys are replaced by Ala and Arg, as was seen in mesophile and thermophile enzymes (Biochemistry, 18:5698-5703, 1979).
glycerol kinase; thermostable enzyme; nucleotide sequence; crystallization; Thermus aquaticus
-15-
Synthesis of Optically Active 1,4-Thiazane-3-carboxylic
Acid via Optical Resolution by Preferential Crystallization
of (RS)-2-Amino-3-[(2-chloroethyl)sulfanyl]propanoic
Acid Hydrochloride
Tadashi SHIRAIWA,・ Kohya TADOKORO, Haruyuki TANAKA, Keiichiro NANBA,
Noriyoshi YOKONO, Katsuyoshi SHIBAZAKI, Motoki KUBO,
and Hidemoto KUROKAWA
Chemical Branch, Faculty of Engineering and Kansai University High Technology Research Center,
Kansai University, Yamate-cho, Suita, Osaka 564-8680, Japan
Received July 16, 1998; Accepted September 1, 1998
Optically active 1,4-thiazane-3-carboxylic acid [TCA] was synthesized from cysteine via optical resolution by||preferential crystallization. The intermediate (RS)-2-ami||||no-3-[(2-chloroethyl)sulfanyl]propanoic acid hydrochlo||ride [(RS)-ACS・HCl] was found to exist as a conglomerate based on its melting point, solubility and IR spectrum. (RS)-ACS・HCl was optically resolved by preferential crystallization to yield (R)- and (S)-ACS・HCl. (R)- and (S)-ACS・HCl thus obtained were recrystallized from a mixture of hydrochloric acid and 2-propanol, taking account of the solubility of (RS)-ACS・HCl, efficiently yielding both enantiomers in optically pure forms. (R)- and (S)-TCA were then respectively synthesized by the cyclization of (R)- and (S)-ACS・HCl in ethanol in the presence of triethylamine.
1,4-thiazane-3-carboxylic acid; 2-amino-3-[(2-chloroethyl)sulfanyl]propanoic acid hydrochloride; conglomerate; optical resolution; preferential crystallization
-16-
A Novel Glycerol Kinase from Flavobacterium meningosepticum:
Characterization, Gene Cloning and Primary Structure
Shin-ichi SAKASEGAWA, Issei YOSHIOKA, Shinzi KOGA, Mamoru TAKAHASHI,
Kunio MATSUMOTO,＊ Hideo MISAKI, and Toshihisa OHSHIMA＊＊
Asahi Chemical Industry Co. Ltd., Ohito-Cho, Tagata-gun, Shizuoka 410-2321, Japan
＊Department of Applied Chemistry, Kanagawa Institute of Technology, Kanagawa 243-0292, Japan
＊＊Department of Biological Science and Technology, Faculty of Engineering, The University of Tokushima, Tokushima 770-8506, Japan
Received July 27, 1998; Accepted September 2, 1998
A thermostable glycerol kinase (FGK) was purified 34-fold to homogeneity from Flavobacterium meningosepticum. The molecular masses of the enzyme were 200 kDa by gel filtration and 50 kDa by SDS-PAGE. The Km for glycerol and ATP were 0.088 and 0.030 mM, respectively. The enzyme was stable at 65°C for 10 min and at 37°C for two weeks. The enzyme gene was cloned into Escherichia coli and its complete DNA was sequenced. The FGK gene consists of an open reading frame of 1494-bp encoding a protein of 498 amino acids. The deduced amino acid sequence of the gene had 40--60％ similarity to those of glycerol kinases from other origins and the amino acid sequence of the putative active site residue reported for E. coli GK is identical to the corresponding sequence of FGK except for one amino acid residue.
glycerol kinase; Flavobacterium meningosepticum; thermostable enzyme; DNA sequence; amino acid sequence similarity
-17-
Substrate Specificity of 2-Deoxy-scyllo-inosose Synthase, the Starter Enzyme
for 2-Deoxystreptamine Biosynthesis, toward Deoxyglucose-6-phosphates
and Proposed Mechanism・
Noriaki IWASE, Fumitaka KUDO, Noriaki YAMAUCHI, and Katsumi KAKINUMA＊
Department of Chemistry, Tokyo Institute of Technology, O-okayama, Meguro-ku, Tokyo 152-8551, Japan
Received July 29, 1998; Accepted August 25, 1998
A crucial enzyme in the biosynthesis of the 2-deoxystreptamine aglycon of clinically important aminocyclitol antibiotics is 2-deoxy-scyllo-inosose synthase (DOIS), which||is responsible for the initial carbocycle formation of||||2-deoxy-scyllo-inosose (1) from D-glucose-6-phosphate||||(G-6-P) (2). To get more insight into the mechanism||||and substrate specificity, deoxy-D-glucose-6-phosphates||||(deoxy-G-6-P) were chemically synthesized and subjected||||to the reaction with DOIS. The enzyme appeared to use||||2-deoxy- and 3-deoxy-G-6-P as substrates, both of which||||were converted into the corresponding dideoxy-scyllo-||||inosose products, but 4-deoxy-G-6-P failed in cyclization||||by DOIS. These results clearly support the proposed reac||||tion mechanism involving the initial oxidation at C-4 of||||the G-6-P substrate. Another implication is the potential||||use of DOIS for the preparation of useful dideoxyinososes.||
||aminocyclitol antibiotic; 2-deoxystreptamine||||biosynthesis; 2-deoxy-scyllo-inosose syn||||thase; deoxyglucose-6-phosphates; dide||oxyinososes
-18-
Cloning of the Gene for Inorganic Pyrophosphatase from a Thermoacidophilic
Archaeon, Sulfolobus sp. Strain 7, and Overproduction of the Enzyme
by Coexpression of tRNA for Arginine Rare Codon
Takayoshi WAKAGI,1,・ Tairo OSHIMA,2 Hiromi IMAMURA,1 and Hiroshi MATSUZAWA1
1Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
2Department of Molecular Biology, Tokyo University of Pharmacy and Life Science, Hachioji,
Tokyo 192-03, Japan
Received August 11, 1998; Accepted August 31, 1998
The gene encoding an extremely stable inorganic pyrophosphatase from Sulfolobus sp. strain 7, a thermoacidophilic archaeon, was cloned and sequenced. An open reading frame consisted of 516 base pairs coding for a protein of 172-amino acid residues. The deduced sequence was supported by partial amino acid sequence analyses. All the catalytically important residues were conserved. A unique 17-base-pair sequence motif was found to be repeated four times in frame in the gene, encoding a cluster of acidic amino acids essential for the function. Although the codon usage of the gene was quite different from that of Escherichia coli, the gene was effectively expressed in E. coli. Coexpression of tRNAArg, cognate for the rare codon AGA in E. coli, however, further improved the production of the enzyme, which occupied more than 85％ of the soluble proteins obatined after removal of heat denatured E. coli proteins.
inorganic pyrophosphatase; thermophile; archaea; gene expression; arginine rare codon
-19-
Note
Transglycosylation of Thiamin by Fungal β-N-Acetylhexosaminidases
Vladimir K・REN,1 Zdenka HU・NKOVA,1 Petr HALADA,1 and Yukio SUZUKI2
1Institute of Microbiology, Laboratory of Biotransformation, Academy of Sciences of the Czech Republic,
Vide・・ nsk´・ a 1083, CZ-142 20 Prague 4, Czech Republic
2Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan
Received February 4, 1998; Accepted July 22, 1998
A new thiamin glycoside, e.g., thiamin β-D-2-deoxy-2-acetamidoglucopyranoside was prepared by transglycosylation by β-N-acetylhexosaminidase from Aspergillus oryzae and characterized spectrally. Series of other fungal β-N-acetyl-hexosaminidases from A. awamori, A. tamari, A. terreus, and Penicillium oxalicum were shown to be able to synthesize the same compound in lower yields.
thiamin; vitamin; β-N-acetylhexosaminidases; transglycosylation; Aspergillus oryzae
-20-
Note
Nutritional Effects of a D-Methionine-containing Solution
on AH109A Hepatoma-bearing Rats
Taizo SASAMURA, Akihiko MATSUDA, and Yukifumi KOKUBA
Infusion Research Department, Medical Information Development Division,
Hoechst Marion Roussel Ltd., 1-3-2 Minamidai, Kawagoe-city, Saitama 350-1165, Japan
Received April 6, 1998; Accepted August 21, 1998
The effects of a D-methionine-containing solution (DMCS) on the nutritional status of AH109A hepatoma-bearing rats receiving total parenteral nutrition were studied. The DMCS solution inhibited the decrease of transferrin in the plasma of tumor-bearing rats when compared with the effect of an L-methionine-containing solution. The survival time was also significantly prolonged in the DMCS-treated rats. These results indicate that DMCS had a beneficial effect on the malnutrition induced in tumor-bearing rats and would be a useful amino acid solution for the nutritional support of cancer patients.
D-methionine; total parenteral nutrition; survival period; nutritional support; hepatoma-bearing rat
-21-
Note
Analysis of the Oligosaccharide Units of Xyloglucans by Digestion
with Isoprimeverose-producing Oligoxyloglucan Hydrolase Followed
by Anion-exchange Chromatography
Teruko KONISHI,・ Yasushi MITSUISHI,＊ and Yoji KATO・・
Laboratory of Food Science, Faculty of Education, Hirosaki University, 1 Bunkyo-cho, Hirosaki 036-8560, Japan
＊National Institute of Bioscience and Human Technology, AIST, 1-1 Higashi, Tsukuba 305-0046, Japan
Received April 27, 1998; Accepted August 18, 1998
A method for rapidly identifying six of the most commonly found xyloglucan oligosaccharide units, XXXG, XLXG, XXLG, XLLG, XXFG, and XLFG was developed by high-performance anion exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) before and after digestion with purified isoprimeverose-producing oligoxyloglucan hydrolase (IPase). Using this method, the compositions of oligosaccharide units of soybean and mung bean xyloglucans were re-examined. Significant amounts of oligosaccharides that have not previously been reported to be oligosaccharide units of soybean and mung bean xyloglucans were found.
xyloglucan; xyloglucan oligosaccharide; isoprimeverose-producing oligoxyloglucan hydrolase
-22-
Note
Cloning of Genomic DNA of Rhizopus niveus Lipase and Expression
in the Yeast Saccharomyces cerevisiae
Mitsutaka KOHNO, Makoto ENATSU, and Wataru KUGIMIYA
Central Research Institute, Tsukuba R＆D Center, Fuji Oil Co., Ltd.,
4-3 Kinunodai, Yawara-Mura, Tsukuba-gun, Ibaraki 300-2497, Japan
Received June 5, 1998; Accepted August 28, 1998
Genomic DNA encoding Lipase I was cloned from Rhizopus niveus strain IFO4759. For expression of this gene in S. cerevisiae, DBY746 was transformed with YEp352PLipS, which had the cloned lipase gene under the control of a PGK promoter. This strain secreted the lipase at a high level (350 U/ml). The strain ND-12B was produced by a mating of DBY746, harboring YEp352PLipS, and NA74-3A, and dissection of asci. This new strain secreted the lipase up to 530 U/ml. Moreover, the lipase was produced most effectively in a medium containing Bacto-yeast extract, soy-peptide, and sucrose.
lipase; Rhizopus niveus; Saccharomyces cerevisiae; secretion
-23-
Note
Simple Synthesis of Both Enantiomers of the C7-C12 Segment of Epothilones
Shinji TANIMORI,＊ Koichi TANIMOTO, and Mitsunori KIRIHATA
Department of Applied Biochemistry, College of Agriculture, Osaka Prefecture University,
1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan
Received June 10, 1998; Accepted August 5, 1998
Both enantiomers of the C7-C12 segment (3 and 4) of antitumor antibiotics epothilones (1and 2) were synthesized from methyl (R)- and (S)-3-hydroxy-2-methylpropionate (5 and 6) in five steps in a fair yield.
epothilone; antitumor antibiotics; segment synthesis; Grignard cross-coupling
-24-
Note
Purification of Pumpkin Glutathione S-Transferase Species Specifically Present
in Cultured Cells Treated by Excessive Concentration
of 2,4-Dichlorophenoxyacetic Acid but Absent in Normal Plants
Masayuki FUJITA,＊ Yasuhiko ADACHI, and Nobuo SAKATO
Department of Plant Sciences, Faculty of Agriculture, Kagawa University,
Miki-cho, Kita-gun, Kagawa 761-0795, Japan
Received June 15, 1998; Accepted September 4, 1998
Pugc is a unique glutathione S-transferase subunit species that is absent in normal pumpkin plants, but GST3c (homodimer of Pugc) accumulates in cultured pumpkin cells treated with 40 ppm 2,4-dichlorophenoxyacetic acid for six days. GST3c was purified by DEAE-cellulose, hydroxylapatite, and S-hexylglutathione-agarose column chromatography, and its homogeneity was confirmed by SDS-PAGE analysis. The specific activity of GST3c was 124 μmol/min/mg protein with 1-chloro-2,4-dinitrobenzene. This was higher than those of the other plant and animal glutathione S-transferases. Mouse antiserum raised against the purified GST3c recognized only Pugc, but not Puga, Pugb, nor Pugd when the reactivity of pumpkin glutathione S-transferase subunits was tested.
glutathione S-transferase; pumpkin; 2,4-dichlorophenoxyacetic acid; purification; antiserum
-25-
Note
Lipase-catalyzed Asymmetric Synthesis of Enantiomerically Pure
(2S,4aS,8S)-4a,8-Dimethyl-2,3,4,4a,5,6,7,8-octahydro-2-naphthalenol
Akira TANAKA,1,・ Takahito TOKUYAMA,2 Akiko SAITO,1 and Takayuki ORITANI2
1Division of Environmental Bioremediation, 2Division of Life Science, Graduate School of Agricultural Science,
Tohoku University, Aoba-ku, Sendai 981-8555, Japan
Received June 16, 1998; Accepted August 7, 1998
Enantiomerically pure (2S,4aS,8S)-(＋)-4a,8-dimethyl-2,3,4,4a,5,6,7,8-octahydro-2-naphthalenol (3), a key intermediate in the synthesis of natural geosmin (1), was prepared by enzymatic kinetic resolution. When racemic 3 was submitted to lipase (PS-30)-catalyzed asymmetric acetylation, employing vinyl acetate as the acyl donor, requisite product (＋)-3 with a high enantiomeric excess was attained as the remaining alcohol. Recrystallization resulted in an enantiomerically pure sample.
lipase; kinetic resolution; asymmetric acetylation; geosmin; musty odour component
-26-
Note
Gassericin A; an Uncommon Cyclic Bacteriocin Produced by Lactobacillus gasseri
LA39 Linked at N- and C-Terminal Ends
Yasushi KAWAI,・ Tadao SAITO, Haruki KITAZAWA, and Takatoshi ITOH
Laboratory of Animal Products Chemistry, Biological Resource Science, Graduate School of Agricultural Science, Tohoku University, Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981-8555, Japan
Received June 17, 1998; Accepted August 4, 1998
Gassericin A, a bacteriocin that was produced by Lactobacillus gasseri LA39, was treated with lysylendopeptidase and 3-bromo-3-methyl-2-(2-nitrophenyl-mercapto)-3H-indole. The fragments were recovered by SDS-PAGE and sequenced. All amino acids of gassericin A were distributed by sequence analysis and the bacteriocin did not contain any modified amino acids. The amino acid sequence of gassericin A completely coincided with that found through the cloning of the structural gene. Gassericin A was shown to be a cyclic bacteriocin (class II) which is bound at the N- and C-terminal ends. This is the first example of a cyclic bacteriocin from lactobacilli lactic acid bacteria.
bacteriocin; Lactobacillus gasseri; gassericin A; acidocin B; cyclic structure
-27-
Note
Promotion by a Peptidyl Growth Factor, Phytosulfokine, of Chlorophyll
Formation in Etiolated Cotyledon of Cucumber
Seiyei YAMAKAWA,1 Yoshikatsu MATSUBAYASHI,2 Youji SAKAGAMI,2
Hiroshi KAMADA,1 and Shinobu SATOH1
1Institute of Biological Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan
2Department of Biological Sciences, Nagoya University, Chikusa, Nagoya 464-01, Japan
Received June 19, 1998; Accepted August 18, 1998
We examined the effects of a peptidyl growth factor, phytosulfokine-α (PSK-α), isolated from conditioned medium and known to induce the proliferation of single mesophyll cells of asparagus, and its derivatives on chlorophyll formation in etiolated cotyledons of cucumber after illumination. The chlorophyll content was increased by PSK-α treatment. Chlorophyll-content increases were not observed following either [2--5]PSK or Tyr-SO3H treatment. There was no difference between the PSK-α treatments and water controls on increasing rate of fresh weights of cotyledons. These results suggest that PSK-α has the effect of elevating the chlorophyll content in the cells.
phytosulfokine; peptidyl growth factor; cucumber; chlorophyll formation
-28-
Note
Inhibition by a Capsaicin Antagonist (Capsazepine) of Capsaicin-induced
Swimming Capacity Increase in Mice
Kyung-Mi KIM, Teruo KAWADA, Kengo ISHIHARA, Kazuo INOUE,
and Tohru FUSHIKI・
Laboratory of Nutrition Chemistry, Division of Applied Life Sciences, Graduate School of Agriculture,
Kyoto University, Kyoto 606-8502, Japan
Received June 19, 1998; Accepted August 12, 1998
We investigated the endurance swimming capacity of mice injected with CAP antagonist (capsazepine). The increase of endurance swimming capacity by the administration of CAP was significantly suppressed by the injection of capsazepine. At the same time, serum adrenaline secretion, which was induced by CAP, was depressed by capsazepine. These findings suggested that the increase in endurance swimming capacity by CAP was mediated by the CAP receptor.
capsazepine; capsaicin; swimming capacity; adrenaline
-29-
Note
Cloning and Nucleotide Sequence of the Gene Encoding a Serine Proteinase
Inhibitor Named Marinostatin from a Marine Bacterium,
Alteromonas sp. Strain B-10-31
Katsushiro MIYAMOTO,1 Hiroshi TSUJIBO,1,・ Yumi HIKITA,1 Kazumi TANAKA,1 Satomi MIYAMOTO,1
Mariko HISHIMOTO,1 Chiaki IMADA,2 Kaeko KAMEI,3 Saburo HARA,3 and Yoshihiko INAMORI1
1Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094, Japan
2Department of Food Science and Technology, Tokyo University of Fisheries,
Minato-Ku, Konan 4-5-7, Tokyo 108-8477, Japan
3Department of Chemistry and Materials Technology, Faculty of Engineering and Design,
Kyoto Institute of Technology, Matsugasaki, Sakyo, Kyoto 606-8585, Japan
Received June 24, 1998; Accepted August 12, 1998
The gene (mstI) encoding a serine proteinase inhibitor named marinostatin from marine Alteromonas sp. strain B-10-31 was cloned and its nucleotide sequence was analyzed. A short open reading frame of 192 bp encoded 63 amino acids with a molecular weight of 6,985. Furthermore, the initial product of marinostatin (marinostatin L) was purified and its amino acid sequence was analyzed. These results indicate that marinostatin is produced as a unique precursor consisting of the mature peptide and the leader peptide for an ATP-binding cassette (ABC) transporter, and furthermore the initial product of marinostatin is dehydrated and processed by proteolysis to give homologous forms of marinostatin.
serine protease inhibitor; ABC transporter; double glycine-type leader peptide; secretion; ester-linkage
-30-
Note
New Antibacterial Diterpenoids from the Sarcodon scabrosus Fungus＊
Hisao SHIBATA,・ Atsushi IRIE, and Yasufumi MORITA
Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University,
8304 Minamiminowa-mura, Nagano 399-4598, Japan
Received June 29, 1998; Accepted August 11, 1998
The structures of new antibacterial diterpenoids that had been isolated from Sarcodon scabrosus were established by chemical and spectral means to be sarcodonin L (2) and M (3), both having the cyathane skeleton. Other antibacterial compounds were identified to be allocyathin B2 (1), sarcodonin G (4) and sarcodonin A (5) by comparing their spectral data with those of authentic samples.
Sarcodon scabrosus; Phylacteriaceae; cyathane diterpenoid; antibacterial activity
-31-
Note
Thermal Isomerization of All-trans-Lutein in a Benzene Solution
Achmad SUBAGIO,1 Naofumi MORITA,1,・ and Shigeo SAWADA2
1Laboratory of Food Chemistry, College of Agriculture, Osaka Prefecture University,
Gakuencho 1-1, Sakai, Osaka 599-8531, Japan
2Faculty of Science and Technology, Kinki University, Higashi-Osaka 577-8502, Japan
Received June 29, 1998; Accepted August 17, 1998
Four major cis-lutein isomers could be identified after thermal isomerization of all-trans-lutein in a benzene solution system; namely, 13′-, 13-, 9′- and 9-cis-luteins. Using both all-trans-lutein and a mixture of these purified cis-luteins as starting materials, the quantitative changes determined by HPLC response show that this thermal isomerization was a reversible reaction. The equilibrium constants (kC) of 13′- and 13-cis-luteins were higher than those of 9′-, and 9-cis-luteins, suggesting that the amount formed of the former isomers was higher than that of the latter.
carotenoid; isomerization; lutein
-32-
Note
Compilation of mRNA Sequences Surrounding the AUG Translation
Initiation Codon in the Green Alga Chlamydomonas reinhardtii
Kazunori IKEDA and Hitoshi MIYASAKA＊,・
Kansai Environmental Engineering Center Co. Ltd, 3-5 Adzuchicho 1-Chome Chuo-ku, Osaka,
Osaka 541-0052, Japan
＊The Kansai Electric Power Co., Technical Research Center, 11-20 Nakoji 3-Chome, Amagasaki,
Hyogo 661-0974, Japan
Received June 30, 1998; Accepted August 27, 1998
Sequences of 118 mRNAs of the green alga Chlamydomonas reinhardtii in the GenBank data base were compiled to examine the consensus sequence surrounding the AUG translation initiation codon. The consensus sequence for C. reinhardtii was found to be gc(A/C)A(A/C)(A/C)・・・AUG・GC. The AUG context of chloroplast proteins (nuclear coded) and non-chloroplast proteins were compared by a separate compilation, and some distinctive features in AUG context of chloroplast proteins were found.
AUG; translation initiation; consensus sequence; green alga; Chlamydomonas
-33-
Note
Novel Cyclohexene Compound from Lasiodiplodia theobromae IFO 31059
Hideyuki MATSUURA, Naomi OBARA, Noriko CHISAKA, Akitami ICHIHARA,
and Teruhiko YOSHIHARA・
Department of Bioscience and Chemistry, Faculty of Agriculture, Hokkaido University,
Sapporo 060-8589, Japan
Received July 9, 1998; Accepted September 1, 1998
A novel cyclohexene compound (1), which is structurally related to theobroxide (2), was isolated from a culture filtrate of the fungus, Lasiodiplodia theobromae IFO 31059. The potato micro-tuber-inducing activity of this compound was observed at a concentration of 10－3 M in the medium, whereas theobroxide (2) showed its activity at 10－5 M.
Lasiodiplodia theobromae; potato micro-tuber-inducing activity; theobroxide; carbonate cyclohexene
-34-
Note
Synthesis of a Novel Phosphate Ester of a Vitamin E Derivative
and Its Antioxidative Activity
Sayuri MIYAMOTO,・・,＊ Takuro KOGA,＊＊ and Junji TERAO・,＊＊＊
＊National Food Research Institute, The Ministry of Agriculture, Forestry and Fisheries,
Tsukuba, Ibaraki 305-0856, Japan
＊＊Noda Institute for Scientific Research, Noda, Chiba 278-0037, Japan
＊＊＊Department of Nutrition, School of Medicine, The University of Tokushima, Tokushima 770-8503, Japan
Received July 16, 1998; Accepted August 20, 1998
A novel phosphate ester containing a chromanol structure was synthesized from 1,2-diacyl-sn-glycero-3-||phospho - 2′ - hydroxyethyl - 2′,5′,7′,8′ - tetramethyl - 6′ - hydro||xychroman (PCh) by hydrolysis catalyzed by phospholipase C from Bacillus cereus. The structure of the product was found by spectral analyses to be 2-(2′,5′,7′,8′-tetramethyl-6′-hydroxychromanyl)ethylphosphate (Ch-P). Ch-P was highly soluble in the aqueous phase at neutral pH values and exerted higher antioxidative activity than α-tocopherol and PCh in the Fe(III)/ascorbic acid-catalyzed peroxidation of a fish oil emulsion and the autoxidation of a rat brain homogenate.
vitamin E; phospholipase C; antioxidant activity; tocopherol; chromanol
-35-
Note
Prevention of Initial Supercooling in Progressive Freeze-concentration
Ling LIU,・ Tomoyuki FUJII, Kiro HAYAKAWA,＊ and Osato MIYAWAKI・・
Department of Applied Biological Chemistry, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku,
Tokyo 113-8657, Japan
＊Research Institute, Kagome Co. Ltd., 17 Nishitomiyama, Nishinasuno-machi, Nasu-gun,
Tochigi-ken 329-2762, Japan
Received July 16, 1998; Accepted August 31, 1998
A physical method is proposed that uses a cooling plate with many small holes to prevent initial supercooling in progressive freeze-concentration, and thus avoid serious contamination of the ice produced. The higher chance for ice nucleation of the water molecules in the holes due to the temperature gradient in the cooling plate resulted in the initial supercooling being completely prevented. Accordingly, the purity of the ice initially formed was substantially improved when compared with that by the standard vessel without holes in the cooling plate.
freeze concentration; supercooling; purity of ice; progressive freezing
-36-
Preliminary Communication
Structural Analysis of Xyloglucan Oligosaccharides by the Post-source
Decay Fragmentation Method of MALDI-TOF Mass Spectrometry:
Influence of the Degree of Substitution by Branched Galactose, Xylose,
and Fucose on the Fragment Ion Intensities
Tohru YAMAGAKI,a,b Yasushi MITSUISHI,a and Hiroshi NAKANISHIa＊
aNational Institute of Bioscience and Human-Technology, AIST, 1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan
bDepartment of Biochemistry, Saitama University, Urawa, Saitama 338-8570, Japan
Received June 22, 1998; Accepted October 7, 1998
Highly branched xyloglucan oligosaccharides were analyzed by the post-source decay (PSD) fragmentation method of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The ratio of [M-Xyl]＋ and [M-Gal]＋ fragment ion intensities could be used to characterize the degree of Gal substitution at the non-reducing end, because the number of possible chemical species was directly related to their relative ion intensity. The intensity of the [M-Fuc]＋ ion was predominantly strong in the fragment spectrum of fucosyl oligosaccharides as the first fragmentation, indicating the fucosyl linkage to be much weaker than the other glycosidic linkages in the MALDI-PSD fragmentation. Setting fragment ion [M-Fuc]＋ to the pseudo precursor ion [MF]＋, the second fragmentation ions were produced from [MF]＋ in the drift region in PSD fragmentation of fucosyl oligosaccharides.
MALDI-TOFMS; post-source decay (PSD) fragmentation; oligosaccharide; xyloglucan; structural analysis
-37-
Preliminary Communication
Identification of a Structural Gene Encoding a Metallothionein-like Domain
that Includes a Putative Regulator Protein for Streptomyces
Protease Gene Expression
Seiichi TAGUCHI,＊ Takahiro OGAWA, Takeshi ENDO, and Haruo MOMOSE
Department of Biological Science and Technology, Science University of Tokyo, 2641 Yamazaki,
Noda-shi, Chiba 278-8510, Japan
Received August 19, 1998; Accepted September 30, 1998
An open reading frame (termed ORF-PR) encoding a metallothionein-like domain-including protein was found upstream of a previously identified Streptomyces chymotrypsin-type protease gene (sam-P20). Promoter and terminator activities of ORF-PR were detected using the promoterless Streptomyces tyrosinase gene as a reporter gene and expression of ORF-PR was supposed to occur before that of sam-P20 gene. Frameshift mutation anaysis showed that the ORF-PR product might act as a repressive regulator of the sam-P20 gene.
Streptomyces; metallothionein-like domain; sequence comparison; DNA-binding motif; protease gene
-38-
Preliminary Communication
Biosynthesis of striatol in cultured cells of liverwort, Ptycanthus striatus
Kenji KATOH,1 Kanae YAMAMOTO,2 Masateru YAMAMOTO,2 Makoto HASHIMOTO,2
and Kensuke NABETA2＊
1Shionogi Abrahi Laboratory, Kohga-cho, Shiga-ken 520-3423, Japan
2Department of Bioresource Science, Obihiro University of Agriculture and Veterinary Medicine,
Obihiro 080-8555, Japan
Received September 1, 1998; Accepted October 11, 1998
NMR analyses of striatol produced by suspension-cultured cells of liverwort, Ptycanthus striatus, in the presence of [2-13C]- and [4,4-2H2]-MVAs confirmed proton-catalyzed cyclization between the distal and central double bonds in FPP and a concerted series of 1,2-migrations of hydrogen and a methyl group, with subsequent elimination of a proton.
liverwort; Ptycanthus striatus; striatol biosynthesis; cyclization; 1,2-migration