Production of Heterologous Providencia rettgeri Penicillin Acylase in Escherichia coli

於大腸菌中進行Prouidencia rettgeri Penicillin Acylase生產之異型基因表現

周志雄(C. P.Chou) ； 林明毅(M. I. Lin) ； 王文昌(W. C. Wang)

《Journal of the Chinese Institute of Chemical Engineers》 31卷2期 (2000/03) Pp. 135-144

https://doi.org/10.6967/JCICE.200003.0135

摘要

本研究為植株Prouidencia rettgeri 之 pac基因，並於大腸桿菌中進行異型基因表現，並探討各種環境因素對其基因表現之影響。

並列摘要

In this work, we constructed several expression plasmids for the production of Pravidencia rettgeri penicillin acylase (EC 3.5.1. 11; PAC) in Escherichia coli. DNA fragments containing the pac gene from P. rettgeri ATCC31O52 were PCR-amplified and cloned in-to the expression vectors so that the pac gene expression was controlled by the lac or trc pro-mater system. The effects of culture conditions, such as IPTG concentration, temperature, and carbon source, on the native or heterologous expression were investigated. Among a selection of expression systems, JM1O9 harboring pUTKnPAC2601 gave the highest PAC activity and could be of interest for industrial application. Cultivation should be perfonned at a temperature ranging from 28°C to 33°C and the medium could be supplemented with glycerol. The host/vector sys-tern offers an opportunity for high-temperature-oriented PAC production, which is usually con-ducted at a low temperature. Volumetric PAC activity at more than fiftyfold (~820 U/L) that of the native expression in ArCC31O52 (~15 U/L) could be reached by optimization of the host/vector system and culture conditions.

並列關鍵字

Heterologous Expression ； Native Expression ； Penicillin Acylase ； Cloning ； Recombinant DNA Technology ； Catabolite Repression