Functional analysis of the C-terminal propeptide of keratinase from Bacillus licheniformis BBE11-1 and its effect on the production of keratinase in Bacillus subtilis |
| |
| Institution: | 1. Synergetic Innovation Center of Food Safety and Nutrition, Wuxi 214122, China;2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;3. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China;4. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;5. School of Biotechnology, Jiangnan University, Wuxi 214122, China;1. Department of Microbiology and Biotechnology, Faculty of Natural Sciences, Vilnius University, M. K. Čiurlionio 21/27, LT-03101 Vilnius, Lithuania;2. Mass Spectrometry Laboratory, Institute of Biochemistry and Biophysics, Polish Academy of Science, Pawinskiego 5a, 02-106 Warsaw, Poland;1. Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia;2. Laboratory of Natural Products Chemistry and Biomolecules (LNPCB), University of Blida, 1, Road of Soumaâ, P.O. Box 270, 09000 Blida, Algeria;3. National Centre for Research and Development of Fisheries and Aquaculture (CNRDPA), 11, Bd Amirouche PO Box 67, Bou Ismaïl, 42415 Tipaza, Algeria;4. National Leather and Shoe Center (CNCC), 17 Road of Leather, Z.I. Sidi Rezig, 2033 Mégrine, Ben Arous, Tunisia;1. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;2. Synergetic Innovation Center of Food Safety and Nutrition, Wuxi 214122, China;3. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;4. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China;5. School of Biotechnology, Jiangnan University, Wuxi 214122, China;1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, PR China;2. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, Jiangsu 214122, PR China;3. School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, PR China |
| |
| Abstract: | The keratinase from Bacillus licheniformis BBE11-1 is a serine protease and expressed as a pre-pro-precursor. To produce a mature and active keratinase, the propeptide must be cleaved on the C-terminal via cis or trans. In this study, to enhance the production of keratinase in Bacillus subtilis, single amino acid substitutions, single residue deletions and linkers were introduced at the C-terminus of the propeptide. The results showed that optimizing the residue of cleavage site of propeptide will affect the cleavage efficiency of propeptide, and the mature enzyme yield of Leu(P1)Ala mutant increases 50% compared with the wild-type. In addition, inserting linkers and deleting individual residues at the C-terminal of the propeptide decreases the mature keratinase production. Our results indicated that the primary structure of the C-terminus of propeptide is crucial for the mature keratinase production. Propeptide engineering at C-terminus may be an effective approach to increase the yield of keratinase. |
| |
| Keywords: | Keratinase Propeptide Cleavage efficiency |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
