嗜盐芽孢杆菌青霉素G酰化酶交联酶聚集体在6-氨基青霉烷酸生产中的应用。
Application of cross-linked enzyme aggregates of Bacillus badius penicillin G acylase for the production of 6-aminopenicillanic acid.
作者信息
Rajendhran J, Gunasekaran P
机构信息
Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, Madurai, India.
出版信息
Lett Appl Microbiol. 2007 Jan;44(1):43-9. doi: 10.1111/j.1472-765X.2006.02043.x.
AIMS
Optimization of 6-aminopenicillanic acid (6-APA) production using cross-linked enzyme aggregates (CLEA) of Bacillus badius penicillin G acylase (PAC).
METHODS AND RESULTS
CLEA-PAC was prepared using purified/partially purified PAC with phenylacetic acid as active-site blocking agent and glutaraldehyde as cross-linker. Conversion of penicillin G to 6-APA by CLEA-PAC was optimized using response surface methodology (RSM) (central composite rotatable design) consisting of a three-factor-two-level pattern with 20 experimental runs.
CONCLUSION
Nearly, 80% of immobilization yield was obtained when partially purified enzyme was used for the preparation of CLEA-PAC. Quantitative conversion of penicillin G to 6-APA was observed within 60 min and the CLEA-PAC was reusable for 20 repeated cycles with 100% retention of enzyme activity.
SIGNIFICANCE AND IMPACT OF THE STUDY
The faster conversion of penicillin G to 6-APA by CLEA-PAC and efficient reusability holds a strong potential for the industrial application.
目的
利用浅黄芽孢杆菌青霉素G酰化酶（PAC）的交联酶聚集体（CLEA）优化6-氨基青霉烷酸（6-APA）的生产。
方法与结果
以苯乙酸为活性位点阻断剂、戊二醛为交联剂，使用纯化/部分纯化的PAC制备CLEA-PAC。采用响应面法（RSM）（中心复合旋转设计）对CLEA-PAC催化青霉素G转化为6-APA的过程进行优化，该设计为三因素两水平模式，共20次实验。
结论
使用部分纯化的酶制备CLEA-PAC时，固定化产率接近80%。在60分钟内可观察到青霉素G向6-APA的定量转化，且CLEA-PAC可重复使用20个循环，酶活性保持100%。
研究的意义和影响
CLEA-PAC能更快地将青霉素G转化为6-APA，且具有高效的可重复使用性，在工业应用方面具有巨大潜力。
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