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Takashi NAKAMURA, Shigeru UTSUMI, Tomohiko MORI
1986 年 50 巻 10 号 p.
2429-2435
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Heat-induced gelation and gel properties in a mixed system of soybean 7S and 11S globulins were studied. The gel hardness changed sigmoidally with the protein concentration and relative proportion of the globulins. The gel turbidity was decreased by the coexistence of 7S globulin. The gelling time was either retarded or shortened depending on the amount of 7S globulin. The 7S and 11S globulins interact noncovalently to form soluble aggregates consisting of both globulins in the course of gel formation. The extent of the interaction was affected by the proportions of the globulins. We conclude that such interactions may be the cause of various behaviors of gelation and gel properties observed here.
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Makoto KAKITANI, Ben'ichiro TONOMURA, Keitaro HIROMI
1986 年 50 巻 10 号 p.
2437-2444
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Valyl-tRNA synthetase (L-valine: tRNA
val ligase (AMP forming); EC 6.1.1.9) (VRS) was purified from
Bacillus stearothermophilus NCA 1503 by streptomycin treatment, ammonium sulfate fractionation, and batch adsorption on DEAE-Sephadex A-50, then column chromatographies on hydroxylapatite, DEAE-Toyopearl 650S, BUTYL-Toyopearl 650S, and Sephacryl S-300 superfine. The enzyme preparation, purified approximately 1100-fold, was homogeneous on polyacrylamide gel disc electrophoresis and consisted of a single chain polypeptide. The molecular weight was about 100, 000 and 110, 000 by SDS-polyacrylamide gel electrophoresis and Sephacryl S-300 gel filtration, respectively. Enzyme activity in tRNA aminoacylation reaction increased as the temperature increased up to 52°C at pH 7.6 and pH 8.5. pH-optima of the aminoacylation activity were 8.5 at 30°C and 8.0 at 52°C under our conditions.
Ligand-induced decrease of the enzyme protein fluorescence was observed and used as a probe for studying the binding of substrates (L-valine and ATP) to VRS. Substrate bindings were also studied by equilibrium dialysis; two moles of L-valine or ATP were bound to a mole of enzyme when each substrate was examined separately. These two binding equilibria of L-valine or ATP seemed equivalent when each substrate is bound alone.
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Satoru KUSAMA, Isao KUSAKABE, Yasuyuki NAKAMURA, Shigeru EDA, Kazuo MU ...
1986 年 50 巻 10 号 p.
2445-2451
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
The enzyme system from
Streptomyces sp. W19-1 formed several kinds of transfer products (TPs) when incubated in the presence of both Stevioside (ST) and curdlan. Three of the major TPs (A, B, and C) were separated and purified using HP-20 column chromatography, gel filtration on TOYOPEARL HW-40F, and preparative high-performance liquid chromatography.
The structures of the three were identified by chemical and enzymatic methods; A is 13-O-β-sophorosyl-19-O-β-laminaribiosyl steviol, B is 13-O-β-3
2-β-glucosylsophorosyl-19-O-β-glucosyl steviol, and C is 13-O-β-sophorosyl-19-O-β-laminaritriosyl steviol. The three were obtained for the first time in a pure state.
抄録全体を表示
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Kazuo YAMAGUCHI, Shuichi ISHINO, Kazumi ARAKI, Kunikatsu SHIRAHATA
1986 年 50 巻 10 号 p.
2453-2459
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
13C NMR spectroscopy was applied to studying lysine biosynthesis in
Corynebacterium glutamicum ATCC 21543, a lysine producing mutant. It was cultured in a medium containing [1-
13C]glucose or [6-
13C]glucose as the sole carbon source and the
13C NMR spectrum of the culture filtrate was measured.
13C labeling patterns of L-lysine produced were well explained by the putative metabolic pathways of the bacterium. Fixation of
13CO
2 liberated from the labeled substrates and the operation of the tricarboxylate cycle in the fermentation were obviously observed. The dual operations of the classical diaminopimelate pathway and the diaminopimelate dehydrogenase bypath were supported. Calculation of the contribution ratios of the metabolic pathways was attempted.
抄録全体を表示
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Nozomi HIURA, Mikihiko KOBAYASHI, Tasuku NAKAJIMA, Kazuo MATSUDA
1986 年 50 巻 10 号 p.
2461-2467
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Two cell wall associated (β1, 3)glucanases in
Neurospora crassa, 1, 3-glucanase-I and II, were solubilized by autolysis of the cell walls with 2M sodium chloride, and purified by laminarioligosaccharide AH-sepharose affinity chromatography followed by preparative disc gel electrophoresis. The 1, 3-glucanase-I has a strict specificity for (β1, 3)glucan and produces o-glucose by an exo-type mode of action. The 1, 3-glucanase-II has specificities for β-glucoside in addition to (β1, 3)glucan. The 1, 3-glucanase-I is more resistant to pH change and metal ions than the 1, 3-glucanase-II.
抄録全体を表示
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Hideo OE, Masaaki HIROSE, Etsushiro DOI
1986 年 50 巻 10 号 p.
2469-2475
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Thiol reagents have been shown to induce the gelation of conalbumin [M. Hirose, H. Oe and E. Doi,
Agric. Biol. Chem.,
50, 59 (1986)]. The gelation mechanisms were investigated in relation to the structure of this protein. Following the addition of 2-mercaptoethanol, the sulfhydryl groups of conalbumin increased almost linearly during the first 50 min of incubation. The relative hydrophobicity measured by fluorescent dyes increased concomitantly with increases in the sulfhydryl groups. The circular dichroism spectra revealed that the thiol reagent induces some changes in the tertiary structure but no prominent alterations in the secondary structure. The transmission electron microscopic study showed that at 20min of incubation conalbumin formed rod-like structures and that at 32 min of incubation these structures produced branched aggregates. The SDS-polyacrylamide gel electrophoresis indicated that intermolecular covalent interactions are not involved in the formation of conalbumin gel.
抄録全体を表示
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Mikio KAWAMORI, Yutaka ADO, Seigo TAKASAWA
1986 年 50 巻 10 号 p.
2477-2482
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Mutants with enhanced β-glucosidase production were isolated by the plate method with 4-methylumbelliferyl-β-D-glucoside as a substrate of β-glucosidase in
Trichoderma reesei. Among them, the mutant strain CDU-11 had the highest production of β-glucosidase. In 5-liter jar fermentor culture of CDU-11 with 6% Avicel as a carbon source, β-glucosidase production was significantly increased by keeping the pH at 4.0 during the first 3 days, then keeping the pH at 5.5 for 4 days. CMCase reached 320 U/ml, Avicelase 29 U/ml, and β-glucosidase 21 U/ml. The ratio of β-glucosidase to CMCase in the CDU-11 culture was about three times that in KY 746, the parent strain. The saccharification of Avicel and alkali-treated bagasse by the cellulolytic system of CDU11 became much more effective than with other strains.
抄録全体を表示
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Michihiro SUGANO, Takahiro ISHIDA, Katsuko YOSHIDA, Kazunari TANAKA, M ...
1986 年 50 巻 10 号 p.
2483-2491
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
The oil produced by the mold
Mortierella ramanniana var.
angulispora IFO 8187, containing approximately 6% of γ-linolenic acid (GLA) and 10% linoleic acid (LA), was examined for its effects on blood cholesterol and eicosanoid levels in rats. When rats were fed cholesterol-enriched diets containing 10% fats, the hypocholesterolemic activity of the mold oil was comparable with that of safflower oil containing 75% LA and was significantly greater than in palm olein containing 16% LA. That the hypocholesterolemic effect could be ascribed exclusively to GLA was confirmed by comparing the effects of ethyl-GLA prepared from the mold oil and ethyl-LA. In rats fed the mold oil the aortic production of PGI
2 tended to be higher, whereas the level of plasma TXB
2 was similar to or lower than those in rats fed safflower oil. The specific activity of Δ
6-desaturase in liver microsomes was significantly higher and the ratio of arachidonate/linoleate in liver phosphatidylcholine was much higher in the mold oil than in safflower oil. These results are indicative of the specific function of the
Mortierella oil in the regulation of eicosanoid production as well as cholesterol metabolism.
抄録全体を表示
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Tomio MORINO, Kenkichi TAKAGI, Tsuneo NAKAMURA, Tomohisa TAKITA, Hiuga ...
1986 年 50 巻 10 号 p.
2493-2497
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Cosmid pR4Cl is a derivative of multicopy plasmid pIJ365 which has an insertion of the
cos (cohesive end site) region of actinophage R4 [T. Morino, H. Takahashi and H. Saito,
Mol. Gen. Genet.,
198, 228 (1985)]. When the donor R4 phage was propagated in
S. lividans carrying the plasmid, the phage lysate contained transducing particles which encapsulated head-to-tail concatemers of the plasmid DNA. These particles could mediate transfer of the plasmid at a high frequency. We examined conditions that gave a maximum transduction frequency of cosmid pR4Cl. Conditions which depress R4 phage propagation, such as incubation of recipient
S. parvulus at a high temperature, improved the frequency. Obviously such conditions minimized the lethal effect of viable phage propogation. The highest transduction frequency obtained so far was around 3×10
-3 transductants per infected phage when
S. lividans was used as the recipient. This was about 30 per cent of the cosmid transducing particles estimated from the cosmid DNA content in the transducing lysate. The significance of cosmid transduction for gene manipulation in
Streptomyces strains is also discussed.
抄録全体を表示
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Hiromichi OHTA, Kazuhiko OZAKI, Gen-ichi TSUCHIHASHI
1986 年 50 巻 10 号 p.
2499-2502
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Reduction of various numbers of 2, 4-alkanediones by fermenting bakers' yeast has been found to be highly regio- and enantioselective, resulting in the formation of 2-hydroxy-4-alkanones. In some cases, optical purities of the products exceeded over 99% as determined by HPLC analysis of MTPA esters. The highest yield was obtained when the reaction was carried out at pH 8 with low substrate concentration. As the representative β-hydroxyketones, the absolute configuration of (+)-2-hydroxy-4-octanone (
2d) and (+)-3-hydroxy-l-phenyl-l-butanone (
2h) was unambiguously determined to be (S) by comparing the specific rotation of diol derivatives with those of authentic specimens.
抄録全体を表示
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Susumu OGUSHI, Makoto ANDO, Daisuke TSURU
1986 年 50 巻 10 号 p.
2503-2507
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Formaldehyde dehydrogenase from
Pseudomonas putida C-83 was found to contain 7 halfcystine residues per subunit monomer, as checked by the method of performic acid oxidation. Approximately 7 sulfhydryl groups per subunil monomer were titrated with 5, 5, -dithiobis(2-nitrobenzoic acid) (DTNB) after denaturation with 8 M urea. In the native enzyme, modification of three sulfhydryl groups per subunit with
p-chloromercuribenzoate (PCMB) led to the complete loss of enzyme activities for both formaldehyde and
n-butanol. Hydrogen-peroxide competitively inhibited the enzyme activity for formaldehyde, while it was only slightly inhibitory to the activity for
n-butanol. Both formaldehyde and hydrogen-peroxide protected one sulfhydryl group per subunit monomer from modification with PCMB. Moreover, hydrogen-peroxide was hardly reactive to the enzyme which was preincubated with formaldehyde.
From these observations, we conclude that one of three PCMB-reactive sulfhydryl groups is essential for the binding of formaldehyde, and hydrogen-peroxide modifies this sulfhydryl group.
抄録全体を表示
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Yukio KIMURA, Noriko YASUDA, Hiroko TANIGAKI-NAGAE, Toshikatsu NAKABAY ...
1986 年 50 巻 10 号 p.
2509-2516
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Acetoacetate decarboxylase is a valuable tool for clinical analysis of ketone bodies in human plasma. After screening many microorganisms, we found
Bacillus polymyxa A-57 to be a new enzyme source with a good yield of acetoacetate decarboxylase. After purifying the intracellular enzyme by three-stage column chromatography, we identified it as a single protein by SDS-polyacrylamide gel electrophoresis. The enzyme had a molecular weight of approximately 280, 000 and consisted of 10 (±2) identical subunits. It had an optimum pH of 5.9 and was stable up to about 60°C for 30min. The apparent
Km and
Vmax values for lithium acetoacetate were 0.94mM and 296μmol/min/mg, respectively. In addition, the decarboxylase activity was found in the broth. After purification, we found that it was due to an active peptide which we named A-57-9, which we identified as the antibiotic polymyxin M
1. However, the
Vmax value (0.25μmol/min/mg) of the peptide was very much lower and the
Km value (400 mM) was higher than those of real acetoacetate decarboxylase.
抄録全体を表示
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Akira YOKOTA, Ken-ichi SASAJIMA
1986 年 50 巻 10 号 p.
2517-2524
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Cell-free extracts of
Bacillus subtilis contain enzyme activities which catalyze an acyloin-type condensation reaction (carboligase reaction) resulting in the formation of l-deoxy-ketoses. The reactions are deduced to proceed as follows:
pyruvate+aldose→CO
2+1-deoxy-ketose (I)
acetoin+aldose→acetaldehyde+1-deoxy-ketose (II)
methylacetoin + aldose→acetone+1-deoxy-ketose (III)
Experiments with mutants of
B. subtilis defective in pyruvate dehydrogenase (PDH) or acetoin dehydrogenase (AccDH) and with partially purified enzyme preparations revealed that PDH (EC 1.2.4.1) catalyzes reaction (I), and AccDH catalyzes reactions (II) and (III).
That the PDH purified from
Escherichia coli and the PDC purified from bovine heart also catalyzed reaction (I) indicates that 1 -deoxy-ketose-forming activities are widely distributed. One of the reactions catalyzed by these enzymes is the formation of 1-deoxy-D-
threo-pentulose, a precursor of biosynthesis of thiazole ring of thiamine.
抄録全体を表示
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Komakazu GOMI, Yasuhide OTA, Yasuji MINODA
1986 年 50 巻 10 号 p.
2525-2530
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Three types of lipase activators (α, β, γ) were isolated from the culture broth of
Saccharomycopsis lipolytica using high performance liquid chromatography. Activator y was the most active for the lipase reaction. One of them (β) was identified with a mixture of 3, 5-dihydroxy-7-tetradecenoic acid and related compounds by the method of NMR and GC-MS analyses. The free carboxyl group in the compounds was essential for the activation of the lipase reaction.
抄録全体を表示
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Komakazu GOMI, Yasuhide OTA, Yasuji MINODA
1986 年 50 巻 10 号 p.
2531-2536
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Lipase activator activated the reaction by
Saccharomycopsis lipolytica lipase at neutral pH in the presence of calcium ions, and 5 μg of the activators were sufficient to cause the reaction to proceed at maximum activity in the presence of 2 μl of tributyrin and 0.4 units of the lipase in a total volume of 360μl.
To define the roles of the activator and calcium ion, we studied interactions between the activator and the lipase, between the activator and a hydrophobic interface, and between the lipase and the interface. Results suggest that the interfacial adsorption of the lipase is the limiting process of lipolysis and that it is controlled by the activator and by the concentration of calcium ions.
抄録全体を表示
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Emi SAKAMOTO, Etsuro SUGIMOTO, Yasuo KITAGAWA
1986 年 50 巻 10 号 p.
2537-2544
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Labeling with
35S-methionine of dispersed hepatocytes prepared from neonatal rat liver and successive immunoprecipitation with antiserum against tyrosine aminotransferase (TAT) indicated that increase of TAT activity to a peak about 12 hours after birth and the decrease thereafter are mainly due to changes of TAT synthesis. Similar changes of TAT activity was also observed in the livers of premature neonates which were taken out by Caesarian section and nursed by foster mothers. This indicated that the appearance of TAT activity at this period is not an event programmed along with fetal development but is triggered by birth. The level of glucagon in neonatal plasma increased after birth. Administration of glucagon to neonates caused a great increase of TAT activity whereas the effect of dexamethasone was not so evident. These suggested that glucagon is an important factor affecting the abrupt appearance of TAT after birth.
抄録全体を表示
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Yoshiki TANI, Byung Dae YOON, Hideaki YAMADA
1986 年 50 巻 10 号 p.
2545-2552
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
A KCN-resistant mutant,
Methylomonas sp. YK 56, contained three kinds of soluble cytochromes
c (cytochromes
c-I,
c-II, and
c-III) though the wild type strain contained two kinds (cytochromes c-I and c-III). The proportion of the three cytochromes c of the mutant were 2.4, 71.5, and 26.1%, and that of the two cytochromes c of the wild type strain were 2.1 and 97.9%, respectively.
Cytochromes
c-II and
c-III of the mutant were purified by a procedure involving ammonium sulfate fractionation and DEAE-, CM-cellulose, and Sephadex G-150 column chromatography. Cytochrome
c-II was obtained as crystals with ammonium sulfate. Both absorption peaks of the α-band of the two cytochromes c were at 551.5 nm at room temperature and the β-band of cytochrome
c-II had a shoulder at 530 nm. Molecular weights of the two cytochromes c were 16, 000 and 20, 000, respectively and their isoelectric points were 4.1 and 3.5, respectively.
抄録全体を表示
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Yasuo KONDO, Masuo NAKANO
1986 年 50 巻 10 号 p.
2553-2559
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Free ceramide and cerebroside were isolated from black gram sprouts of all germinating stages. Free ceramide and cerebroside were found to increase during germination.
The major sphingosine bases of free ceramide were 4-hydroxysphingenine and 4-hydroxysphinganine (trihydroxy type) while that of cerebroside was sphinga-4, 8-dienine (dihydroxy type). A change in the component sphingosine base was that 4-hydroxysphingenine in free ceramide and cerebroside increased slightly after germination.
The major fatty acid of free ceramide was α-hydroxylignoceric acid while that of cerebroside was α-hydroxypalmitic acid. Changes in component fatty acid were that α-hydroxylignoceric acid in both sphingolipids increased after germination.
抄録全体を表示
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Rhyuji FURUTA, Shigeo KAWATA, Shunsuke NARUTO, Akira MINAMI, Shozo KOT ...
1986 年 50 巻 10 号 p.
2561-2572
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
The acylated, amidated and esterified derivatives of
N-acetylglucosaminyl-β(1→4)-
N-acetylmuramyl tri- and tetrapeptide were synthesized and examined as to their protective effect on pseudomonal infection in the mouse and pyrogenicity in the rabbit. Modifications of the terminal end function of the peptide moieties in their molecules caused enhancement of resistance to pseudomonal infection and reduction of pyrogenicity. Among the compounds tested, sodium
N-acetylglucosaminyl-β(1→4)-
N-acetylmuramyl-L-alanyl-D-isoglutaminyl-(L)-stearoyl-(D)-
meso-2, 6-diaminopimelic acid-(D)-amide and sodium
N-acetylglucosaminyl-β(1→4)-
N-acetylmuramyl-L-alanyl-D-isoglutaminyl-(L)-stearoyl-(D)-
meso-2, 6-diaminopimelic acid-(o)-amide-(L)-Dalanine were found to be advantageous and conceivably worthwhile for further investigation as immunobiologically active compounds.
抄録全体を表示
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Kazuaki NISHIKAWA, Susumu OI
1986 年 50 巻 10 号 p.
2573-2577
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
The bacterium, which was isolated from soil and identified as
Enterobacter sp., was induced by hexachlorophen (HCP) and chlorhexidine (CH), as well as benzalkonium chloride (BC), to produce acidic polysaccharide. HCP is a bisphenol and CH is a bisbiguanido, while BC is a quarternary ammonium compound. The cells produced the maximum amount of the polysaccharide (0.3-0.9mg as total sugar/mg dry weight cells) in a 0.07M potassium phosphate buffer (pH 7.2) containing 0.22 M glucose and approximately 0.1 mm BC or HCP, or 0.06 mM CH. There was no growth of the cells in these conditions. The polysaccharides produced in the presence of each drug were all composed of fucose, glucose, galactose and glucuronic acid. At the optimum concentration for polysaccharide production, a large amount of UV-absorbing material was released from the cells.
抄録全体を表示
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Koji UCHIDA, Shunro KAWAKISHI
1986 年 50 巻 10 号 p.
2579-2583
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Oxidative depolymerization of several polysaceharides induced by oxygen radical generating systems such as ascorbic acid-copper ion was investigated by the increases in their reducing powers and by gel-filtration. All polysaceharides used in these experiments were greatly depolymerized within 24 hr at room temperature. We thought that the main factors in the oxidation of these polysaceharides were the actions of hydroxyl radical (•OH) generated from the autoxidation of ascorbic acid.
On the other hand, dextrans having molecular weights above 10
3-10
4 were more susceptible to the oxidation reaction with •OH than smaller oligosaccharides (MW<10
3), and we estimated that the formation of sugars-copper ion complexes was highly important to the oxidative depolymerization of polysaceharides.
抄録全体を表示
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Mikihiko KOBAYASHI, Ikuko YOKOYAMA, Kazuo MATSUDA
1986 年 50 巻 10 号 p.
2585-2590
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Graphical analysis of inhibition kinetics for dextransucrase from
Leuconostoc mesenteroides was done with typical inhibitors, competitive and noncompetitive. Based on the plots of Yonetani-Theorell and Semenza-Balthazar, mutual competition between the pairs of inhibitors of identical kinetic type was observed, while combination of competitive and noncompetitive inhibitors gave no significant mutual interactions. By the procedure of Nitta
et al., binding sites for competitive and noncompetitive inhibitors were shown to be distant from each other. Moreover, two noncompetitive inhibitors competed with each other for a single binding site on the enzyme. Although biphasic reciprocal plots may suggest rather complicated binding of various inhibitors, the results obtained by the three graphical methods are fully explained when competitive and noncompetitive inhibitors for substrate sucrose bind to the so-called donor- and acceptor-sites of dextransucrase, respectively.
抄録全体を表示
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Katsuji WATANABE, Eiji TANIGUCHI
1986 年 50 巻 10 号 p.
2591-2595
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Sixty-two phenylacetic acid derivatives having phenylpropane skeleton were synthesized. Their physiological effects on the growth of lettuce seedlings were distinguished from those of plant hormones and an antiauxin. The derivatives retarded the growth in a similar manner to the auxin transport inhibitors, NPA and CPD. A few derivatives including 2-[3'-(
m-chlorophenyl)isoxazol-5'-yl]phenylacetic acid were up to 10 times as potent as CPD in inhibition of the hypocotyl growth and IAA transport. The phenylacetic acids did not cause, even at high concentrations, any serious damage such as a herbicidal would. Their remarkable growth-retarding effects may be due, at least in part, to their ability to prevent the polar movement of IAA within tissues.
抄録全体を表示
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Hiroshi TAKAGI, Yasushi MORINAGA, Kiyoshi MIWA, Shigeru NAKAMORI, Kono ...
1986 年 50 巻 10 号 p.
2597-2603
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Versatile plasmid vectors useful for gene cloning in
Brevibacterium lactofermentum, a glutamic acid-producing bacterium, have been constructed. The trimethoprim (Tp)-resistant dihydrofolate reductase gene derived from chromosomal DNA of the Tp-resistant mutant of
B. lactofermentum was introduced into pAM330, a cryptic plasmid in
B. lactofermentum. The constructed cloning vector pAJ228 (7.6kb) exists in 10 to 20 copies in cells of
B. lactofermentum and donated Tpresistance, which is a useful selective marker of transformants. pAJ228 was further improved to a versatile plasmid vector pAJ224 having some profitable characteristics such as smaller size (3.7 kb), higher copy number (60-80 copies), and additional useful cloning sites (
BamHI,
PstI and
SalI) equipped with two different promoters arranged at both orientations for the expression of passenger DNA without promoter. These plasmids were stably retained in
B. lactofermentum even in the absence of Tp over many generations. Thus, they have been found very powerful vectors for gene cloning in
Brevibacterium and the related bacteria.
抄録全体を表示
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Michiko MORI, Isamu SHIIO
1986 年 50 巻 10 号 p.
2605-2614
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Phosphoenolpyruvate (PEP) carboxylase purified from
Brevibacterium flavum was specifically activated by fructose 1, 6-bisphosphate (FBP). The other intermediates of sugar metabolism or their structural analogues did not influence the activity. FBP decreased the apparent
Km for PEP but did not affect that for another substrate, bicarbonate, or the apparent maximum velocity for PEP. The dissociation constants for FBP from enzyme-FBP and enzyme-PEP-FBP complex were 63 and 32 μM, respectively, being almost equivalent to those for acetyl-CoA. Synergistic activation by FBP and acetyl-CoA was not observed with the
B. flavum enzyme, unlike the
Escherichia coli enzyme. FBP, like acetyl-CoA, was kinetically competitive with aspartate. With respect to another feedback inhibitor, 2-oxoglutarate, acetyl-CoA was non-competitive, whereas FBP was of mixed-type,
i.e., FBP but not acetyl-CoA prevented 2-oxoglutarate from binding to the enzyme to a certain extent. Homotropic cooperativity was observed only with FBP but not with acetyl-CoA in the absence of inhibitors. Cooperativities of FBP and acetyl-CoA were increased by aspartate but not by 2-oxoglutarate. In the aspartate-overproducing mutant enzyme, the Michaelis constant for PEP was decreased, whereas the inhibitor constant for aspartate with or without simultaneous addition of 2-oxoglutarate and the activator constants for FBP and acetyl-CoA were increased. The decreased Michaelis constant for PEP was comparable to the apparent
Km of the wild-type enzyme for PEP in the presence of the saturated concentration of FBP, and would result in a further decrease in the affinity of the mutant enzyme for aspartate.
抄録全体を表示
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Yasuyoshi SAKAI, Yoshiki TANI
1986 年 50 巻 10 号 p.
2615-2620
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
Formaldehyde production was investigated with cells of a mutant, AOU-1, of a methanol yeast,
Candida boidinii S2 grown in methanol-limited chemostat culture. The highest productivity was shown with cells from the culture at a dilution rate of 0.075 hr
-1, when cells had the highest activity of alcohol oxidase and almost minimum activity of formaldehyde dehydrogenase. Under optimal reaction conditions, 950 mM formaldehyde was produced in 10-hr reaction with the cells. By the chemostat culture, not only formaldehyde productivity but also cell productivity was improved in comparison with batch culture. A maximum cell productivity of 0.2 g•liter
-1•hr
-1 and a cell yield of 47% were obtained.
抄録全体を表示
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Yoshimitsu YAMAZAKI, Hidekatsu MAEDA
1986 年 50 巻 10 号 p.
2621-2631
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
An enzyme that reduces benzoylformate with NADH to form (
R)-mandelate was extracted from cells of
Streptococcus faecalis IFO 12964 and purified to more than 95% purity as evidenced by gel electrophoresis. Physicochemical and enzymic properties were studied. From the substrate specificity, we concluded that the enzyme was a kind of (
R)-2-hydroxyisocaproate dehydrogenase. Optically pure (
R)-(-)-mandelic acid was prepared with the enzyme, NADH, and alcohol, formate or glucose dehydrogenase in 84-93% yield. Five (
R)-2-hydroxyalkanoic acids (C
4-C
6) or their Ba salts, (
R)-(+)-3-phenyllactic acid and (
S)-(-)-3-chlorolactic acid were also prepared with the enzyme.
抄録全体を表示
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Mutsuo KANNO
1986 年 50 巻 10 号 p.
2633-2635
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Masao YOSHIZAWA, Mitsuyoshi UEDA, Sabiha MOZAFFAR, Atsuo TANAKA
1986 年 50 巻 10 号 p.
2637-2638
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Tsutomu KANEKO, Hideki SUZUKI, Jsuyoshi TAKAHASHI
1986 年 50 巻 10 号 p.
2639-2641
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Kikuo SHIMBO
1986 年 50 巻 10 号 p.
2643-2645
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Kazutaka MIYATAKE, Haruhiko SAKURABA, Shozaburo KITAOKA
1986 年 50 巻 10 号 p.
2647-2649
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Kazutaka MIYATAKE, Koji WASHIO, Akiho YOKOTA, Yoshihisa NAKANO, Shozab ...
1986 年 50 巻 10 号 p.
2651-2653
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Hiroshi OHASHI, Tetsu TSURUSHIMA, Tamio UENO, Hiroshi FUKAMI
1986 年 50 巻 10 号 p.
2655-2657
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Teruyoshi MATOBA, Kiyozo HASEGAWA, Keisuke KITAMURA, Makoto KITO
1986 年 50 巻 10 号 p.
2659-2661
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Toshio TANAKA, Gaysorn DHAVISES, Makoto TANIGUCHI, Susumu OI
1986 年 50 巻 10 号 p.
2663-2665
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Horace G. CUTLER, Patsy D. COLE, Richard F. ARRENDALE, Ronald L. BASSF ...
1986 年 50 巻 10 号 p.
2667-2668
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Mitsuru MONMA, Keiji KAINUMA
1986 年 50 巻 10 号 p.
2669-2671
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Takahiro TOBA, Susumu ABE, Keizo ARIHARA, Susumu ADACHI
1986 年 50 巻 10 号 p.
2673-2674
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Eiko MATSUMURA, Satoshi YOMODA, Eiko YAMAMOTO, Tatsu KAWANO, Takashi S ...
1986 年 50 巻 10 号 p.
2675-2677
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Kazuhiro IRIE, Hideo HAYASHI, Motoo ARAI, Koichi KOSHIMIZU
1986 年 50 巻 10 号 p.
2679-2680
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Hideki TATEBA, Satoru MIHARA
1986 年 50 巻 10 号 p.
2681-2683
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Mikie HAYASHIDA, Kenji WATANABE
1986 年 50 巻 10 号 p.
2685-2687
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Yoshiki KONO, Takeshi KINOSHITA, Setsuo TAKEUCHI, J. M. DALY
1986 年 50 巻 10 号 p.
2689-2691
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Michio MURATA, Masaki SANO, Takashi IWASHITA, Hideo NAOKI, Takeshi YAS ...
1986 年 50 巻 10 号 p.
2693-2695
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー
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Masahiro NISHII, Yasuko FUJIHARA, Jin-ichiro INAGAKI, Kazuo OGAWA, Fuj ...
1986 年 50 巻 10 号 p.
2697-2699
発行日: 1986年
公開日: 2006/04/05
ジャーナル
フリー