Using inexpensive substrate to achieve high-level lipase A secretion by Bacillus subtilis through signal peptide and promoter screening |
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| Affiliation: | 1. Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin, 300072, People’s Republic of China;2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People’s Republic of China;3. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People’s Republic of China;4. Department of Fermentation Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People’s Republic of China;5. School of Biological Engineering, Dalian Polytechnic University, Dalian, 116034, People’s Republic of China;6. National Engineering Laboratory for Industrial Enzymes, Tianjin, 300308, People’s Republic of China;1. Molecular and Nanomedicine Research Unit, Centre for Nanoscience and Nanotechnology, Sathyabama Institute of Science and Technology, Chennai 600119 Tamil Nadu India;2. Department of Biotechnology, Alagappa University, Science Campus, Karaikudi 630003, Tamil Nadu India;1. College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, Zhejiang, China;2. The Third Affiliated Hospital of Qiqihar Medical College, Qiqihar, Heilongjiang, China;1. College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China;2. Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China;1. School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China;2. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China;3. Suqian Industrial Technology Research Institute of Jiangnan University, Suqian, 223814, China;1. Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Kunming 650500, China;2. School of Life Science, Yunnan Normal University, Kunming 650500, China |
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| 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. |
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| Keywords: | Lipase A Signal peptide Promoter Chicken feather hydrolysate Glycerol |
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