Institute of Bioengineering, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China.
World J Microbiol Biotechnol. 2013 Mar;29(3):431-40. doi: 10.1007/s11274-012-1195-y. Epub 2012 Oct 21.
In this study, nitriles were used as sole sources of nitrogen in the enrichments to isolate nitrile-converting microorganisms. A novel fungus named ZJB-09150 possessing nitrile-converting enzymes was obtained with 3-cyanopyridine as sole source of nitrogen, which was identified by morphology, biology and 18S rDNA gene sequence as Fusarium proliferatum. It was found that F. proliferatum had ability to convert nitriles to corresponding acids or amides and showed wide substrate specificity to aliphatic nitriles, aromatic nitriles and ortho-substituted heterocyclic nitriles. The nitrile converting enzymes including nitrilase and nitrile hydratase in ZJB-09150 were induced by ε-caprolactam. Nitrilase obtained in this study showed high activity toward 3-cyanopyridine. It was active within pH 3.0-12.0 and temperature ranging from 25 to 65 °C with optimal at pH 9.0 and temperature 50-55 °C. The enzyme was thermostable and its half-life was 12.5 and 6 h at 45 and 55 °C, respectively. Under optimized reaction conditions, 60 mM 3-cyanopyridine was converted to nicotinic acid in 15 min, which indicated ZJB-09150 has potentials of application in large scale production of nicotinic acid.
在这项研究中,腈被用作富集物中氮的唯一来源,以分离能够转化腈的微生物。以 3-氰基吡啶为唯一氮源,获得了一株具有腈转化酶的新型真菌,命名为 ZJB-09150,通过形态学、生物学和 18S rDNA 基因序列鉴定为多枝木霉。研究发现,多枝木霉具有将腈转化为相应酸或酰胺的能力,并且对脂肪族腈、芳香族腈和邻位取代杂环腈表现出广泛的底物特异性。ZJB-09150 中的腈转化酶包括腈酶和腈水合酶,可被ε-己内酰胺诱导。本研究获得的腈酶对 3-氰基吡啶表现出较高的活性。它在 pH3.0-12.0 和 25-65°C 的温度范围内具有活性,最佳 pH 值为 9.0,最佳温度为 50-55°C。该酶具有热稳定性,在 45 和 55°C 下的半衰期分别为 12.5 和 6 h。在优化的反应条件下,60mM 的 3-氰基吡啶在 15 分钟内转化为烟酸,表明 ZJB-09150 具有在烟酸的大规模生产中应用的潜力。