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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2012, Vol. 32 Issue (09): 41-47    
研究报告     
高产(+)γ-内酰胺酶菌株的筛选与发酵产酶研究
陈红干, 倪晔, 孙志浩
江南大学工业生物技术教育部重点实验室 江南大学生物工程学院 无锡 214122
Isolation and Fermentation Optimization of a (+)γ-Lactamase-producing Strain
CHEN Hong-Gan, NI Ye, SUN Zhi-Hao
Key Lab of Industrial Biotechnology, Ministry of Education, Jiangnan University, School of Biotechnology, Wuxi 214122, China
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摘要： 从土壤中筛选获得高产(+)γ-内酰胺酶的微生物菌株,并鉴定和保藏为Delftia sp. CGMCC No. 5755。对该Delftia sp菌株的发酵产酶条件进行了研究,结果表明,最适发酵培养基为:蔗糖30 g/L,蛋白胨30 g/L,牛肉膏25 g/L,乙酰胺5 g/L,MgSO4 1 g/L;最适发酵温度及初始pH分别为32℃和pH 7.0。该菌株在上述条件下发酵培养20 h,菌体生物量为16.0 g/L,(+)γ-内酰胺酶的酶活为692 U/L。采用Delftia sp.静息细胞对100 g/L的外消旋底物2-氮杂二环--庚烷-5-烯-3-酮(简称(±)γ-内酰胺)的水解拆分反应中,产物(-)γ-内酰胺光学纯度大于99.9%e.e.,转化率为53.7%。研究为生物催化法高效制备光学纯(+)γ-内酰胺提供了可行的途径。
关键词： 2-氮杂二环-[2.2.1]-庚烷-5-烯-3-酮((±)γ-内酰胺);  Delftia sp.;  CGMCC No. 5755;  (+)γ-内酰胺酶;  静息细胞;  生物拆分    
Abstract: A (+)γ-lactamase-producing microorganisms with high enantioselectivity was isolated from soil, and was identified and deposited as Delftia sp.CGMCC No.5755. The optimized fermentation medium consists of: 30 g/L sucrose, 30 g/L peptone, 25 g/L beef extract, 5 g/L acetamide, and 1 g/L MgSO4; and the optimum temperature and initial pH are 32℃ and pH 7.0, respectively. Under the optimized conditions, 15.6 g/L of Delftia cells and 687 U/L of (+)γ-lactamase were obtained after 20 h of fermentation. Biotransformation using Delftia resting cells was carried out at 100 g/L of (±)γ-lactam substrate, the optical purify of product (-)γ-lactam reached over 99.9%e.e., and the conversion rate was 53.7% after 12h.The study provides a feasible approach for the biocatalytic preparation of (-)γ-lactam.
Key words: Racemic 2-azabicyclo[2.2.1]hept-5-en-3-one[(±)γ-lactam]    Delftia sp.    CGMCC No. 5755    (+)γ-lactamase    Resting cell    Bioresolution
收稿日期: 2012-06-08 出版日期: 2012-09-25
ZTFLH:  TQ 920.6  
基金资助: 国家"973"计划(2011CB710800);教育部新世纪优秀人才支持计划 (NCET-11-0658);江苏省自然科学基金(BK2011150);高等学校学科创新引智计划 (111-2-06)资助项目
通讯作者: 倪晔     E-mail: yni@jiangnan.edu.cn
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引用本文:

陈红干, 倪晔, 孙志浩. 高产(+)γ-内酰胺酶菌株的筛选与发酵产酶研究[J]. 中国生物工程杂志, 2012, 32(09): 41-47.

CHEN Hong-Gan, NI Ye, SUN Zhi-Hao. Isolation and Fermentation Optimization of a (+)γ-Lactamase-producing Strain. China Biotechnology, 2012, 32(09): 41-47.

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https://manu60.magtech.com.cn/biotech/CN/    或    https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I09/41

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