title: Expression and display of a novel thermostable esterase from Clostridium thermocellum on the surface of Bacillus subtilis using the CotB anchor protein.
abstract: Esterases expressed in microbial hosts are commercially valuable, but their applications are limited due to high costs of production and harsh industrial processes involved. In this study, the esterase-DSM (from Clostridium thermocellum) was expressed and successfully displayed on the spore surface, and the spore-associated esterase was confirmed by western blot analysis and activity measurements. The optimal temperature and pH of spore surface-displayed DSM was 60 and 8.5 °C, respectively. It also demonstrates a broad temperature and pH optimum in the range of 50-70, 7-9.5 °C. The spore surface-displayed esterase-DSM retained 78, 68 % of its original activity after 5 h incubation at 60 and 70 °C, respectively, which was twofold greater activity than that of the purified DSM. The recombinant spores has high activity and stability in DMSO, which was 49 % higher than the retained activity of the purified DSM in DMSO (20 % v/v), and retained 65.2 % of activity after 7 h of incubation in DMSO (20 % v/v). However, the recombinant spores could retain 77 % activity after 3 rounds of recycling. These results suggest that enzyme displayed on the surface of the Bacillus subtilis spore could serve as an effective approach for enzyme immobilization.
authors: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, Jiangsu Province, China. hyc@ujs.edu.cn.|National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 10090, China. hyc@ujs.edu.cn.|Division of Biological Sciences, University of California at San Diego, La Jolla, CA, 92093-0116, USA. hyc@ujs.edu.cn.
authors: Huayou
authors: H
authors: Chen
authors: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, Jiangsu Province, China.
authors: Tianxi
authors: T
authors: Zhang
authors: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, Jiangsu Province, China.
authors: Jinru
authors: J
authors: Jia
authors: Division of Biological Sciences, University of California at San Diego, La Jolla, CA, 92093-0116, USA.
authors: Ake
authors: A
authors: Vastermark
authors: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, Jiangsu Province, China.
authors: Rui
authors: R
authors: Tian
authors: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, Jiangsu Province, China.
authors: Zhong
authors: Z
authors: Ni
authors: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, Jiangsu Province, China.
authors: Zhi
authors: Z
authors: Chen
authors: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, Jiangsu Province, China.
authors: Keping
authors: K
authors: Chen
authors: Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, Jiangsu Province, China.
authors: Shengli
authors: S
authors: Yang
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D048013:Acetivibrio thermocellus / Q000201:enzymology* / Q000235:genetics; D004121:Dimethyl Sulfoxide; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000235:genetics / Q000378:metabolism; D004950:Esterases / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D013171:Spores, Bacterial / Q000378:metabolism; D013696:Temperature; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; C439051:CotB protein, Bacillus subtilis; D004800:Enzymes, Immobilized; D004950:Esterases; D004121:Dimethyl Sulfoxide
keywords: Bacillus subtilis; CotB; Esterase; Surface display; Thermophilic