胆固醇诱导甾短杆菌发酵产胆固醇氧化酶能力的研究

食品科学 ›› 2012, Vol. 33 ›› Issue (21): 253-257.

胆固醇诱导甾短杆菌发酵产胆固醇氧化酶能力的研究

傅裕，丁晓雯*，蒋丽施，李凯，罗金凤，任美燕

西南大学食品科学学院，重庆市农产品加工技术重点实验室

收稿日期:2011-09-25 修回日期:2012-09-23 出版日期:2012-11-15 发布日期:2012-11-09
通讯作者: 丁晓雯 E-mail:xiaowend2001@yahoo.com

Optimization of Cholesterol-Induced Cholesterol Oxidase Production by Brevibacterium sp.

Received:2011-09-25 Revised:2012-09-23 Online:2012-11-15 Published:2012-11-09
Contact: xiaowen Ding E-mail:xiaowend2001@yahoo.com

摘要/Abstract

摘要： 利用胆固醇诱导甾短杆菌发酵来提高胆固醇氧化酶产量；通过对单因素的研究，选取影响比较显著的3个因素：胆固醇质量浓度、吐温-80添加量、超声波功率，并利用Box-Behnken试验设计和响应曲面分析法进一步研究得到结果：甾短杆菌产酶的最佳条件是当超声时间为20min时，胆固醇质量浓度为3.30g/L、吐温-80添加量为3.72mL/L、超声波功率为80W，在此条件下测得的胆固醇氧化酶产量为731.967U/L。比未使用胆固醇诱导产酶时，酶产量提高了1.17倍。

关键词: 胆固醇氧化酶, 胆固醇, 吐温-80, 超声波功率

Abstract: In this study, Brevibacterium sp. was cultured in a fermentation medium supplemented with cholesterol to enhance the production of cholesterol oxidase. Through one-factor-at-a-time analysis, three experimental conditions including cholesterol and Tween-80 concentration in the fermentation medium and ultrasonic power (for promoting the release of intracellular cholesterol oxidase) were found to significantly influence the activity of crude cholesterol oxidase solution. The optimal cholesterol and Tween-80 concentration as well as ultrasonic power and treatment time were 3.30 g/L, 3.72 mL/L, 80 W and 20 min, respectively, as optimized using Box-Behnken experimental design and response surface methodology. The resulting cholesterol oxidase activity was 731.967 U/L, which was increased by 1.17-fold when compared with that obtained in the absence of cholesterol.

Key words: cholesterol oxidase, cholesterol, Tween-80, ultrasonic power

中图分类号:

TS201.3

傅?裕，丁晓雯--*，蒋丽施，李?凯，罗金凤，任美燕. 胆固醇诱导甾短杆菌发酵产胆固醇氧化酶能力的研究[J]. 食品科学, 2012, 33(21): 253-257.

参考文献

[1] LEBOVICS V K,MAGDOLNA A,ODON G.Enzymatic determination of cholesterol oxidase[J].J Sci Food Agric,1996,71:21~26 .
[2]MACLACHLAN J,WOTHERSP.OON A T L,et al.Cholesterol oxidase:source,physical properties and analytical applications[J].Steriod Biochem Molecular Biology,2007,72:169-195.
[3] VOLKER F W, ALBERT A D, HERMANN S, et al. Quantitative determination of metabolic fluxes during coutilization of two carbon sources: comparative analysis with Corynebacterium glutamicum during growth on acetate and glucose [J].Journal of Biotechnology.2000,3088-3096.
[4]CORBIN D R,GREENPLATE J T,WONG E Y,et al.Cloning of an insecticidal cholesterol oxidase gene and its expression in bacteria and in plant protoplasts[J].Applied and Environmental Microbiology,1994,60(12):4239-4244.
[5]PURCELL J P, GREENP T. Cholesterol oxidase: a potent insecticidal p rotein active against boll weevil larvae [J]. Biochem Biophys Res Commun,1993,196 (3):1406-1413.
[6] SUZUKI K.Antiobesity agent 4-Cholesten-3-one,US. 5610152 [P].1995-1-11.
[7] SUZUKI K.4-eholesten-3-one for the control of obesity.World Patent.1993,12:794-798.
[8] KASHIMA M,KINOSHITA T,INAOKA A,et al.Cholestanones for treatment of liver diseases,JP. 0769898[P],1995-03-14.
[9]CHRISTODOULOU S,HUNG T V,TREWHELL M A ,et al. Journal of Food Protect,1994(57):908-912.
[10] 赵芳.胆固醇氧化酶的研究进展[J].食品科技,2008,12(5):50-52.
[11] 许晓燕，张锐，谭晓晶等.胆固醇氧化酶的发酵条件及酶学性质研究[J].食品发酵与工业,2006,32(2):41-45.
[12]季文明，陈毅力，张和春等.比色法测定胆固醇氧化酶酶活[J].无锡轻工大学学报,2000,19(3):251-254.
[13]钟学敏，孙艳，张玲等.胆固醇氧化酶分离纯化及应用的研究进展[J].生物技术通报,2009,4:44-49.
[14] 杨胜利，张慧，石轩峰等.响应曲面分析应用于短杆菌产酶工艺的优化[J].中国粮油学报,2004,19(6):60-62.
[15] 杨胜利.超声波促进胆固醇的分散对短杆菌产酶的影响[J].工业微生物,2005,35(4):33-35.
[16] CHEN F L,YI X T, LONG G,et al.Bioconversion of yolk cholesterol by extracellular cholesterol oxidase from Brevibacterium sp..Food Chemistry[J].2002,77:457-463.
[17]刘翠平.响应面法优化干酪乳杆菌LC2W合成胞外多糖的培养条件[J].食品发酵与工业,2008,34(12):85-89.