对用于全细胞生物催化剂的米根霉菌株产酶条件的优化及脂肪酶的酶学性质进行了研究。结果表明，在250 mL三角瓶液体培养中，米根霉适宜的产酶条件为：氮源为2%胰蛋白胨与0.1%NaNO3复合，碳源为0.3%的大豆油，培养基初始pH=5.5，培养温度28℃，培养时间约58 h。酶学性质研究表明，所产脂肪酶的最适pH=7.5，最适温度为35℃，该酶在pH为7~9、温度低于40℃的范围内表现稳定，Mg2+对酶活力有一定促进作用。
Abstract
The wholecell biocatalyst is a novel technology for biodiesel production. It can not only overcome the needs of highenergy and environmental pollution in traditional chemical process, but also avoid the high cost and short lifetime of enzymes in immobilizedenzyme catalyzing system. Therefore, the wholecell biocatalyst attracts extensive attention in recent years. In this paper the production conditions of lipase from Rhizopus oryzae were optimized and its enzymatic properties were studied. The results indicated that the suitable conditions for lipase production from R.oryzae were as the follows: nitrogen source was 2% tryptone and 0.1% NaNO3, carbon source was 0.3% soybean, initial pH was 5.5, incubation temperature was 28℃, and incubation time was 58 h, respectively. Furthermore, the properties of the lipase were also characterized. The optimal pH and temperature of lipase for catalysis were 7.5 and 35℃. The lipase was relatively stable in pH range of 7-9 and at 40℃, and Mg2+ had a stimulative effect on the lipase activity.
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1]李香春. 脂肪酶的研究进展[J]. 肉类工业,2003(4):45-48.
[2]颜兴和,王栋,徐岩. 根霉脂肪酶的研究进展[J]. 工业微生物,2005,35(3):45-49.
[3]Jaeger K E, Eggert T. Lipase for biotechnology[J]. Curr Opin Biotechnol, 2002, 13: 390-397.
[4]李燕,潘运国,连毅. 微生物脂肪酶催化及其性质研究进展[J]. 粮食与油脂,2005,10:15-17.
[5]谭天伟,王芳,邓立,等. 生物柴油及应用[J]. 新材料工业,2004,123(2):41-44.
[6]Fukuda H, Kondo A, Noda H. Biodiesel fuel production by transesterification of oils[J]. Journal of Bioscience and Bioengineering, 2001, 92(5): 405-416.
[7]杨继国,林炜铁,吴军林. 酶法合成生物柴油的研究进展[J]. 化工环保,2004,24(2):37-42.
[8]杨学昊,谭天伟. 固定化根霉发酵生产脂肪酶[J]. 生物工程学报,2004,20(2):284-286.
[9]Ban K, Kaieda M, Matsumoto T, et al. Whole cell biocatalyst for biodiesel fuel production utilizing Rhizopus oryzae cells immobilized within biomass support particles[J]. Biochemical Engineering Journal, 2001, 8(1): 39-43.
[10]李迅,李治林,杨杨,等. 制备生物柴油用脂肪酶产生菌的筛选和诱变[J]. 林产化学与工业,2007,27(6):85-89.
[11]钱存柔,黄仪秀. 微生物学实验教程[M]. 北京:北京大学出版社,1999.
[12]李建武. 生物化学实验原理与方法[M]. 北京:北京大学出版社,1994.
[13]Salleh A B, Musani R, Basri M, et al. Extraand intracellular lipases from a thermophilic Rhizopus oryzae and factors affecting their production[J]. Canadian Journal of Microbiology, 1993, 39: 978-981.
[14]Essamri M, Deyris V, Comeau L. Optimization of lipase production by Rhizopus oryzae and study on the stability of lipase activity in organic solvents[J]. Journal of Biotechnology, 1998, 60: 97-103.
[15]王蓉,王远亮,陈国平,等. 米根霉发酵生产L(+)乳酸研究进展[J]. 重庆大学学报:自然科学版,2004,27(12):95-97.
基金
收稿日期：2008-11-03修回日期：2009-09-01基金项目：国家林业局“948”项目(2009-4-63)；国家高技术研究发展计划(2009AA05Z437)；国家林业局“948”创新项目(2006-4-C05); 江苏省研究生科研创新计划项目(164030146)作者简介：李治林(1982—)，博士生。*王飞(通讯作者)，教授，研究方向为生物质能源与化学品、天然产物化学。Email: feiwang@njfu.com.cn。引文格式：李治林,李迅,王丽丽,等. 米根霉产脂肪酶的条件及酶学性质[J]. 南京林业大学学报:自然科学版,2009,33(6):103-107.
