Citation: Jiaxin Gao, Xiaoyang Ou, Pei Xu, Minhua Zong, Wenyong Lou. Cloning, overexpression, and characterization of a novel organic solvent-tolerant lipase from Paenibacillus pasadenensis CS0611[J]. Chinese Journal of Catalysis, ;2018, 39(5): 937-945. doi: 10.1016/S1872-2067(18)63033-5 shu

Cloning, overexpression, and characterization of a novel organic solvent-tolerant lipase from Paenibacillus pasadenensis CS0611

Jiaxin Gao ^a, ,
Xiaoyang Ou ^a ,
Pei Xu ^a ,
Minhua Zong ^b ,
Wenyong Lou ^a,b

a Laboratory of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China;
b State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou 510640, Guangdong, China

Received Date: 20 November 2017
Revised Date: 19 January 2018

Fund Project: This work was supported by the National Natural Science Foundation of China (21336002, 21376096, 21676104) and the Program of State Key Laboratory of Pulp and Paper Engineering (2017ZD05).

We found a novel lipase gene in the Paenibacillus pasadenensis CS0611 strain. The lipase gene sequence was cloned into the pET-28a expression vector to construct a recombinant lipase protein containing 6×His tags at the C-and N-termini, respectively. High-level expression of the lipase in E.coli BL21 (DE3) was obtained upon induction with IPTG at 20℃. The recombinant lipase activity was approximately 1631-fold higher than the wild type. His-tagged recombinant lipase was purified rapidly and efficiently by using Ni-charged affinity chromatography with 63.5% recovery and a purification factor of 10.78. The purified lipase was stable in a broad range of temperatures and pH values, with the optimal temperature and pH being 50℃ and 7.0, respectively. Its activity was stimulated to different degrees in the presence of metal ions such as Ca²⁺, Mg²⁺, and some non-ionic surfactants. In addition, the purified lipase was activated by a series of water-miscible organic solvents such as some short carbon chain alcohols and was highly tolerant to some water-immiscible organic solvents.
- Lipase,
- Paenibacillus pasadenensis CS0611,
- Expression,
- Characterization,
- Organic solvent tolerant
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