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31.
Takayuki Uwajima Hitoshi Yagi Shizuko Nakamura Osamu Terada 《Bioscience, biotechnology, and biochemistry》2013,77(10):2345-2350
Among about 500 strains tested, a newly isolated soil bacterium, Brevibacterium sterolicum nov. sp. KY 3463 (ATCC 21387) showed the highest potency in production of 3β-hydroxysteroid oxidase in the culture fluid.The 3β-hydroxysteroid oxidase was purified from the culture filtrate by a procedure involving ammonium sulfate fractionation, DEAE-cellulose and hydroxyapatite column chromatographies and Sephadex G–75 gel filtration. Crystals of the enzyme were obtained from solutions of the purified preparation by the addition of ammonium sulfate. The crystals appeared as fine rods, with a bright yellow color.The enzyme is homogeneous by disc gel electrophoresis and ultracentrifugation. Sedimentation velocity yields a value of . It exhibits a typical flavoprotein spectrum of absorption maxima at 280, 390, and 470 mμ. 相似文献
32.
Takayuki Uwajima Hitoshi Yagi Osamu Terada 《Bioscience, biotechnology, and biochemistry》2013,77(6):1149-1156
3β-Hydroxysteroid oxidase (3β-hydroxysteroid: oxygen oxidoreductase, EC 1.1.3.6.) from the culture supernatant of Brevibacterium sterolicum ATCC 21387 has a molecular weight of 32,500 and an isoelectric point of 8.9. The enzyme contained 258 amino acid residues and the composition revealed a distinctive feature of a relatively high amount of proline and the absence of alanine and tryptophan. The crystalline enzyme exhibited an absorption spectrum characteristic of a flavoprotein with absorption maxima at 280, 390, and 470 nm with a shoulder at 490 nm. Anaerobic addition of dehydro-epi-androsterone as well as sodium dithionite to the enzyme produced a disappearance of the peaks at 390 and 470 nm. The flavin moiety of the enzyme was isolated and identified as flavin adenine dinucleotide, 1 mole of which was found per mole of protein. The enzyme is sulfhydryl dependent and was inactivated by silver and mercury compounds. Analysis of the enzyme protein by atomic absorption spectrophotometry failed to detect any significant quantity of heavy metals.Various 3β-hydroxysteroids were oxidized and the relative rates of the oxidation were cholesterol, 100; dehydro-epi-androsterone, 41; pregnenolone, 22; and β-sitosterol, 20. The oxidation product of cholesterol by the enzyme was crystallized and identified as 4-cholesten-3-one by melting point, elementary analysis, optical rotation, UV, IR and NMR spectra. The oxidation of cholesterol proceeded as follows: The enzyme would be used for some analytical and preparative purposes in the field of steroid chemistry, e.g., microdetermination of cholesterol in serum. 相似文献
33.
Shunya Takahashi Takayuki Oritani Kyohei Yamashita 《Bioscience, biotechnology, and biochemistry》2013,77(10):2711-2718
The total synthesis of ( + )-methyI phaseate (2b) and its epimer (25) is described. The known β- ketoester (8), which was prepared from ( — )-/f-pinene (4), was converted to a key intermediate (5) via a 1, 4-dioxoester (7). The reaction of 5 with a lithium reagent of the acetylene TBDMS ether (6) in THF-HMPA at — 70°C afforded the desired acetylene alcohol (17) and its epimer (18) in high yields. 17 was transformed into ( + )-methyl phaseate (2b). From this synthetic work, the absolute configuration of natural ( — )-phaseic acid (2a) was confirmed. 相似文献
34.
Takayuki Aoyama Yasukazu Nakakita Masahira Nakagawa Heiichi Sakai 《Bioscience, biotechnology, and biochemistry》2013,77(9):2369-2371
Asymmetric hydrolysis of (dl)-1-acyloxy-2-halo-1-phenylethanes by lipoprotein lipase Amano P from Pseudomonas fluorescens and the lipase from Chromobacterium viscosum afforded the optically active (R) residual substrates and (S)-2-halo-1-hydroxy-1-phenylethanes in 100% enantiomeric excess (e.e.). The length of acyl residues from acetyl to octanoyl in the substrates did not influence the enantioselectivity.Both enantiomers of optically active styrene oxides were synthesized from the enzymatic products. 相似文献
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So called ambreinolal (IV),* a component of ambergris, was first synthesized by CrO3 oxidation of ambreinolol (III)* which was obtained from ambreinolide (II) by reduction with LiAlH4. Ambreinolol (III) was converted to the C17-saturated oxide (VII) in a good yield through the monotosylate (VIII) by treatment with p-toluenesulfonyl chloride in pyridine.Ambrein (I), a major constituent of ambergris, was easily converted to ambrein-tetrahydropyranylether (II), of which thermal decomposition gave back ambrein (I). The tetrahydropyranylether (II) was oxidized to ambreinolal-tetrahydropyranylether (V) in two steps. Ambreinolal-tetrahydropyranylether (V) was synthesized from ambreinolol (VII) in four steps and converted to the C17-unsaturated oxide (VI) on heating. 相似文献
39.
Paknisa Sirimontree Natchanok Sritho Yuka Kanda Shoko Shinya Takayuki Ohnuma 《Bioscience, biotechnology, and biochemistry》2013,77(12):2014-2021
Enhancing the transglycosylation (TG) activity of glycoside hydrolases does not always result in the production of oligosaccharides with longer chains, because the TG products are often decomposed into shorter oligosaccharides. Here, we investigated the mutation strategies for obtaining chitooligosaccharides with longer chains by means of TG reaction catalyzed by family GH18 chitinase A from Vibrio harveyi (VhChiA). HPLC analysis of the TG products from incubation of chitooligosaccharide substrates, GlcNAcn, with several mutant VhChiAs suggested that mutant W570G (mutation of Trp570 to Gly) and mutant D392N (mutation of Asp392 to Asn) significantly enhanced TG activity, but the TG products were immediately hydrolyzed into shorter GlcNAcn. On the other hand, the TG products obtained from mutants D313A and D313N (mutations of Asp313 to Ala and Asn, respectively) were not further hydrolyzed, leading to the accumulation of oligosaccharides with longer chains. The data obtained from the mutant VhChiAs suggested that mutations of Asp313, the middle aspartic acid residue of the DxDxE catalytic motif, to Ala and Asn are most effective for obtaining chitooligosaccharides with longer chains. 相似文献
40.
Phyu Phyu Khine Zar Satoshi Yano Kozue Sakao Fumio Hashimoto Takayuki Nakano Makoto Fujii 《Bioscience, biotechnology, and biochemistry》2013,77(10):1731-1737
Fresh loquat leaves have been used as folk health herb in Asian countries for long time, although the evidence supporting their functions is still minimal. This study aimed to clarify the chemopreventive effect of loquat tea extract (LTE) by investigating the inhibition on proliferation, and underlying mechanisms in human promyelocytic leukemia cells (HL-60). LTE inhibited proliferation of HL-60 in a dose-dependent manner. Molecular data showed that the isolated fraction of LTE induced apoptosis of HL-60 as characterized by DNA fragmentation; activation of caspase-3, -8, and -9; and inactivation of poly(ADP)ribose polymerase. Moreover, LTE fraction increased the ratio of pro-apoptotic Bcl-2-associated X protein (Bax)/anti-apoptotic myeloid cell leukemia 1 (Mcl-1) that caused mitochondrial membrane potential loss and cytochrome c released to cytosol. Thus, our data indicate that LTE might induce apoptosis in HL-60 cells through a mitochondrial dysfunction pathway. These findings enhance our understanding for chemopreventive function of loquat tea. 相似文献