Studies on Biologically Active Substances Formed by Bacilli

The products of several Bacillus strains were investigated on rabbit serum calcium decreasing, oxytocic and toad heart function promoting activities. These products were obtained from the clear supernatant fluid of the culture medium after the cells were removed by centrifugation.

For the production of rabbit serum calcium decreasing substance, Bacillus subtilis K and Bacillus natto No. 8 were found to be usefull, Bacillus megaterium KM, Bacillus cereus var. mycoides and Bacillus subtilis K produced oxytocic principle. Bacillus subtilis K, Bacillus brevis and Bacillus megaterium KM also produced toad heart function promoting factor.

A procedure was developed to obtain the electrophoretically homogenous rabbit serum calcium decreasing substance from culture filtrate of Bacillus subtilis K. The crude substance was obtained as isoelectric precipitate by adjusting the culture filtrate to pH 3.0. The crude substance was purified by gel filtration on a Sephadex G-75 column, isoelectric fractionation and chromatography on DEAE-cellulose column. The purified preparation was shown to be homogenous by Tiselius electrophoresis but was separated into two bands by polyacrylamide electrophoresis. The chemical analysis of this biologically active substance indicated this substance to be a lipoprotein. The substance decreased rabbit serum calcium level about 12% at 6~8hr after intravenous injection (dose; 0.5 mg/kg body weight).     

On the Physiology of Rhizopus Species

Part. V. The preliminary research on the occurrence of zymase and carboxylase in Rhizopus species.

Part. VI. The verification of the occurrence of zymase in Rhizopus species.

Part. VII. On the formation of ethyl alcohol from acetic acid by acetone-Rhizopus (Rhizopus treated by acetone.)

Part. VIII. On the formation of ethyl alcohol from malic acid by Rhizopus species.

Part. IX. On the formation of ethyl alcohol from malic acid by ace’one-Rhizopus.     

Catabolism of Substituted Benzoic Acids by Streptomyces Species

Four thermotolerant actinomycetes from soil, identified as Streptomyces albulus 321, Streptomyces sioyaensis P5, Streptomyces viridosporus T7A, and Streptomyces sp. V7, were grown at 45°C in media containing either benzoic acid or hydroxyl- and methoxyl-substituted benzoic acids as the principal carbon sources. Benzoic acid was converted to catechol; p-hydroxybenzoic, vanillic, and veratric acids were converted to protocatechuic acid; and m-hydroxybenzoic acid was converted to gentisic acid. Catechol, protocatechuic acid, and gentisic acid were cleaved by catechol 1,2-dioxygenase, protocatechuate 3,4-dioxygenase, and gentisate 1,2-dioxygenase, respectively. Dioxygenases appeared only in induced cultures. m-Hydroxybenzoic, m-anisic, and p-anisic acids were gratuitous inducers of dioxygenases in some strains. One strain converted vanillic acid to guaiacol.     

Studies on Amino Acids

The enzymatic resolution of acetyl-dl-methonine previously reported was studied further, in detail. As a result, it was found that metal ion plays an important role on the asymmetric hydrolysis of acyl-dl-methonines by the enzyme preparations of Aspergillus oryzae.     

产朊假丝酵母利用不同有机酸作碳源的研究

分别通过不同有机酸为惟一碳源,研究了产朊假丝酵母（Candida utilis）能够利用的有机酸的种类,旨在为微生物的基础研究和应用基础研究提供部分依据。分别通过对培养时间和菌体在不同有机酸合成培养基中OD值进行统计分析,发现培养时间和柠檬酸、乳酸、琥珀酸、L-苹果酸培养基的OD值极显著相关（P〈0.01）;培养时间和乙酸、酒石酸、富马酸和草酸培养基的OD值无相关性。通过对比菌体培养前后有机酸合成培养基pH值的变化发现乙酸、酒石酸、富马酸和草酸合成培养基的pH值没有明显变化,而苹果酸、乳酸、琥珀酸和柠檬酸为碳源的合成培养基的pH值均明显增大。从而说明产朊假丝酵母能利用L-苹果酸、乳酸、琥珀酸、柠檬酸为碳源,而不能利用乙酸、酒石酸、草酸、富马酸为碳源。     

Isolation and Preliminary Characterization of Hydroxamic Acids Formed by Nitrogen-Fixing Azotobacter chroococcum B-8

The free-living diazotroph Azotobacter chroococcum B-8 responded to iron-limited growth conditions by forming hydroxamic acids and an 85,000-dalton outer membrane protein. The Fe(III)-binding hydroxamate compounds stimulated the growth of Arthrobacter flavescens JG-9 and gave a positive Csaky reaction for bound hydroxylamines. The hydroxamates were isolated from liquid cultures by benzyl alcohol extraction and purified by size exclusion chromatography and high-performance liquid chromatography. Four high-performance liquid chromatography fractions, designated A, B, C, and D, had the characteristic hydroxamate absorption maximum at 420 to 423 nm, which did not shift over a pH range from 3.0 to 9.0. Cyclic voltammograms of the iron-hydroxamate complexes exhibited reduction potentials of −0.426 to −0.442 V for fractions A, B, and D and of −0.56 V for fraction C versus the normal hydrogen electrode at pH 8.0. Based on mass spectra, nominal molecular weights of 800 and 844 were assigned to ferrated compounds A and B, respectively. Reductive hydrolysis of compounds A and B in 57% hydriodic acid yielded ornithine as detected by gas chromatography-mass spectrometry. All of these physiological and chemical data strongly support the hypothesis that the high-affinity iron-binding compounds isolated from A. chroococcum B-8 are hydroxamic acids and probably function as siderophores for this diazotroph.     

Studies on Amino Acids XII.

Although acylase activities in animal tissues, molds, bacteria and yeast have been clarified, nothing has been reported on the acylase activity in plants. Therefore, as a part of a series of studies intending to elucidate the biological significance of the presence of acylase activity, activity in plants was investigated.

As a result, the occurrence of acylase activity in plants such as vegetables, potatoes, pulses, cereals and fruits, has been confirmed for the first time. Besides the above higher plants a relatively higher acylase activity was also found in mushroom.     

Studies on Amino Acids. II

The enzymatic procedures for the resolution of dl-lysine such as asymmetric synthesis of acyl l-lysinc anilide and acyl dl-lysines have been studied. As a result, the procedure consisting in the enzymatic asymmetric hydrolysis of ε-benzoyl-α-acctyl-dl-lysine was found to be the most advantageous for the resolution of dl-lysine.     

静息细胞法对根霉12#产抗生物质机理的研究

利用静息细胞法对中国根霉12#菌株所产生的一种能抑制芽孢杆菌生长的物质的代谢机理进行了研究,结果表明一些碳源类物质对这种抑制剂的产生有一定的促进作用,而且这种抑制剂的合成与菌体蛋白合成系统有关。     

Simulation Studies on the Stabilities of Aggregates Formed by Fibril-Forming Segments of α-Synuclein

Abstract

We performed molecular dynamics simulations for various oligomers with different β-sheet conformations consisting of α-Synuclein 71–82 residues using an all atom force field and explicit water model. Tetramers of antiparallel β-sheet are shown to be stable, whereas parallel sheets are highly unstable due to the repulsive interactions between bulky and polar side chains as well as the weaker backbone hydrogen bonds. We also investigated the stabilities of double antiparallel β-sheets stacked with asymmetric and symmetric geometries. Our results show that this 12 amino acid residue peptide can form stable β-sheet conformers at 320K and higher temperatures. The backbone hydrogen bonds in β-sheet and the steric packing between hydrophobic side chains between β-sheets are shown to give conformational stabilities.