Heterologous expression and characterization of an unsaturated glucuronyl hydrolase from Alteromonas sp. A321.

Autor: Xiao R; College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China., Du C; College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China., Li H; School of Nursing, Qingdao University, Qingdao 266011, China., Zhang M; College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China., Wu Y; College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China., Xing L; College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China., Bu K; College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China., Wang P; College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China. Electronic address: pengwang@ouc.edu.cn.
Jazyk: angličtina
Zdroj: International journal of biological macromolecules [Int J Biol Macromol] 2024 Dec; Vol. 282 (Pt 4), pp. 137012. Date of Electronic Publication: 2024 Oct 31.
DOI: 10.1016/j.ijbiomac.2024.137012
Abstrakt: Strong promoters and stable mRNAs are essential for the overproduction of heterologous proteins in Bacillus subtilis. To improve the strength of natural promoters and ensure robust protein output, promoter and genetic insulator engineering have been used. A series of plasmids containing single and dual promoters and genetic insulators to express alt3796 were engineered, which encoded an unsaturated glucuronyl hydrolase (UGL). As a first step, we screened the host and deleted the signal peptide (SP ALT ) of alt3796, successfully expressed secreted ALT3796 from B. subtilis WB800. Subsequently, to improve expression, we screened the dual promoter P Hag-spoVG from a collection of 22 promoters, which yielded higher enzymatic activity. Finally, using a recombinant strain carrying a plasmid with the P Hag-spoVG dual promoter and a genetic insulator, we obtained 40.9 U/mL of activity. Purified recombinant ALT3796 exhibited good stability and specifically degraded ulvan. In conclusion, a system for the heterologous expression of ALT3796 was constructed, and the obtained protein exhibited favorable properties, suggesting its potential for preparing novel ulvan oligosaccharides.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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