Characterization and overproduction of a thermo-alkaline pectate lyase from alkaliphilic Bacillus licheniformis with potential in ramie degumming |
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| Institution: | 1. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;2. National Engineering Lab for Industrial Enzymes, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;1. Department of Chemical-Biotechnology Engineering, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran;2. Chemical Engineering Department, Engineering Faculty, Razi University, Kermanshah, Iran;3. Research Center of Oils and Fats, Kermanshah University of Medical Science, Kermanshah, Iran;1. Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing 100084, PR China;2. College of Chemistry and Life Science, Shenyang Normal University, Shenyang 110034, PR China;3. Zhejiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, PR China;1. Bioengineering College, Chongqing University, Chongqing 400044, China;2. Research Center for Tobacco Bioengineering and Technology, Chongqing Science and Technology Commission, Yubei District, Chongqing 401147, China;1. Laboratory of Pharmaceutical Engineering, School of Pharmaceutics Science, Jiangnan University, Wuxi 214122, China;2. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;3. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China;1. Department of Protein Chemistry and Technology, CSIR-Central Food Technological Research Institute, Mysuru 570020, India;2. Department of Studies in Biochemistry, University of Mysore, Mysuru, India;1. State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China;2. Agricultural Bio-resource Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350003, PR China |
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| Abstract: | A thermo-alkaline pectate lyase (BliPelA) gene from an alkaliphilic Bacillus licheniformis strain was cloned and overexpressed in Escherichia coli. Mature BliPelA exhibited maximum activity at pH 11 and 70 °C, and demonstrated cleavage capability on a broad range of substrates such as polygalacturonic acid, pectins, and methylated pectins. The highest specific activity, of 320 U mg?1, was towards polygalacturonic acid. Significant ramie (Boehmeria nivea) fiber weight loss (21.5%) was obtained following enzyme treatment and combined enzyme-chemical treatment (29.3%), indicating a high ramie degumming efficiency of BliPelA. The total activity of recombinant BliPelA reached 1450.1 U ml?1 with a productivity of 48.3 U ml?1 h?1 under high-cell-density cultivation with a glycerol exponential feeding strategy for 30 h in 1-l fed-batch fermenter, and 1380.1 U ml?1 with a productivity of 57.5 U ml?1 h?1 after 24 h under constant glucose feeding in a 20-l fermenter using E. coli as the host. The enzyme yields reached 4.5 and 4.3 g l?1 in 1-l and 20-l fed-batch fermenters, respectively, which are higher than those of most reported alkaline Pels. Based on these promising properties and high-level production, BliPelA shows great potential for application in ramie degumming in textile industry. |
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| Keywords: | Alkaline pectate lyase Characterization Overproduction Ramie degumming |
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