低温脂肪酶的产酶条件优化及其酶学性质

中国生物工程杂志

2010, Vol. 30

Issue (03): 67-73

研究报告

低温脂肪酶的产酶条件优化及其酶学性质

苑博华¹,蔡宇杰¹,廖祥儒^1**,恽丽红¹,吴会广¹,张大兵²

1.江南大学工业生物技术教育部重点实验室无锡 214122
2.江苏汉邦科技有限公司淮安 223001

Optimization of Fermentation Condition for a Cold-active Lipase and Enzymatic Characterization

1.The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China
2.Jiangsu Hanbon Science & Technology Co. Ltd, Huai’an 223001, China

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摘要：

利用单因素筛选和正交试验对Burkholderia sp. SYBC LIP-Y发酵产酶的液体培养基和发酵条件进行了优化，其优化配方为：可溶性淀粉10 g/L、牛肉膏15 g/L、NaNO3 0.252 g/L、橄榄油40ml/L、Triton x-100 10ml/L、初始pH 7.5、接种量10%(V/V)，脂肪酶酶活达到85.23U/ml，是优化前的3.63倍。通过对双水相纯化得到的脂肪酶进行酶学性质研究，确定该酶反应的最适pH为10.0，最适温度为30℃，40℃下保温60min酶活性还有80%以上，该脂肪酶为低温脂肪酶，热稳定性好，具有一定的耐醇性，应用前景广阔。

关键词： 低温脂肪酶; 优化; 酶学性质

Abstract:

The medium and condition of liquid fermentation for the yield of lipase of Burkholderia sp. SYBC LIP-Y were optimized with single factor experiment and orthogonal design. The most suitable medium was formulated with the ingredients: 10 g/L tragantine, 15 g/L beef extract, 0.252 g/L NaNO3, 40 ml/L olive oil, 10 ml/L Triton x-100, pH 7.5, and 10% (V/V) of the inoculum size. The lipase activity reached 83.85 U/ml, which was 3.63 folds of that before optimization. Properties of the lipase after purification by aqueous two-phase system was studied. The optimum pH and temperature of the purified enzyme were 10.0 and 30℃ respectively. About 80% of the original activity is maintained by heating at 40℃ for 60 minutes. The lipase belonged to the cold-active lipase. Good temperature stability and resistant to alcohol were proved. There had wide application fields that can be believed.

Key words: Cold-active lipase Optimization Enzymatic property

收稿日期: 2009-09-15 出版日期: 2010-03-25

基金资助:

长江学者和创新团队发展计划（IRT0532）资助项目

通讯作者: 廖祥儒 E-mail: liaoxiangru@163.com

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引用本文:

苑博华蔡宇杰廖祥儒恽丽红吴会广张大兵. 低温脂肪酶的产酶条件优化及其酶学性质[J]. 中国生物工程杂志, 2010, 30(03): 67-73.

YU Bo-Hua, CA Yu-Jie, LIAO Xiang-Ru, YUN Li-Gong, TUN Hui-An, ZHANG Da-Bing. Optimization of Fermentation Condition for a Cold-active Lipase and Enzymatic Characterization. China Biotechnology, 2010, 30(03): 67-73.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I03/67

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