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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2013, Vol. 33 Issue (7): 36-42    DOI:
    
The Optimized Phospholipase A1 Gene Expression of Serratia marcescens PL-06 in E. coli
SU Yan-nan, XUE Zheng-lian, CHEN Tao, MA Qi-ya
Institute of Biologic & Chemical Engineering of Anhui Polytechnic University, Microorganism Fermentation Engineering and Technology Research Center of Anhui Province, Wuhu 241000, China
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Abstract  The phopholipase A1 gene from Serratia marcescens PL-06 was named plaA, and the phospholipase A1 with accessory protein was plaB. The genes plaA and plaB were cloned, ligased with pET-28 a(+)and transferred to BL21(DE3). The engineered stains expressing plaA and plaB was obtained, and named AP28 and BP28. The phospholioase A1 expression level reached peak when the IPTG concentration was 0.2mmol/L, the OD600 was 0.5, the inducing temperature was 37℃ and the inducing time was 4 hours. Relatively the PLA1 protein expression level of AP28 rose from 32% to 46%, and the enzyme activity of refolded inclusion body enhanced from 10.8 U/ml to 12 U/ml. Contrast to BP28, the expression of target protein was low toxic to the host bacterial and easy to purify. Therefore, it is feasible that phospholipase A1 gene plaA expressed in the form of a large number of inclusion bodies by optimizing the inducing conditions, and thereby obtain higher activity of phospholipase A1 and avoid toxicity to the host cell, and it could obtain large amount of phospholipase A1 protein for the subsequent studies.

Key words： Serratia marcescens      Phospholipase A1      Induced expression      Inclusion bodies refolding      Cell toxicity     
Received: 04 December 2012      Published: 25 July 2013
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SU Yan-nan
XUE Zheng-lian
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MA Qi-ya
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SU Yan-nan, XUE Zheng-lian, CHEN Tao, MA Qi-ya. The Optimized Phospholipase A1 Gene Expression of Serratia marcescens PL-06 in E. coli. China Biotechnology, 2013, 33(7): 36-42.

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https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2013/V33/I7/36

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