Using inexpensive substrate to achieve high-level lipase A secretion by Bacillus subtilis through signal peptide and promoter screening. (December 2020)

• Record Type: Journal Article
• Authors: Wu, Fengyi; Ma, Jiayuan; Cha, Yaping; Lu, Delin; Li, Zhiwei; Zhuo, Min; Luo, Xiaochun; Li, Shuang; Zhu, Mingjun
• Is Part Of: Process biochemistry. Volume 99(2020)
• Page Start: 202
• Page End: 210
• Publication Date: 2020-12
• Subjects: Lipase A -- Bacillus subtilis -- Signal peptide -- Promoter -- Chicken feather hydrolysate -- Glycerol; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2020.08.010
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Identification of favorable signal peptide SPlipA for lipase A secretion. Exploration of dual-promoter (P43 -Phag ) for efficient lipase A production. High-level extracellular production of lipase A (710 U/mL) by B. subtilis . Excellent performance (1165 U/mL) using glycerol & feather hydrolysate as substrate. Abstract: Lipase A exhibits great commercial value due to its applications in the food and paper industry, pharmaceutical chemistry and household chemicals. In this study, we first conducted a comparison of different signal peptides and promoters for the construction of the productive lipase A strain. The maximum extracellular lipase activity was identified as 64.9 U/mL after 12-h fermentation at 37 °C by B. subtilis L25 in which the lipase gene was led by SPlipA and controlled by P43 -Phag . Another aim was to determine whether supplementing a culture medium with a combination of glycerol and Streptomyces sp. SCUT-3 digested chicken feather hydrolysates was beneficial to lipase production. Systematic optimization experiments were designed and carried out. Finally, a satisfactory lipase level of 1164.9 U/mL was accomplished in Terrific-Broth medium at a C/N ratio of 0.5 ( v/v ). This work demonstrates the feasibility of B. subtilis L25 for lipase A industrial production using glycerol and microbes treated chicken feather hydrolysates as inexpensive carbon and nitrogen source.