title: High-level extracellular production of alkaline polygalacturonate lyase in Bacillus subtilis with optimized regulatory elements.
abstract: The present work aims to construct a robust recombinant Bacillus subtilis to achieve secretory production of alkaline polygalacturonate lyase (PGL). First, 6 signal peptides (amyX, bpr, vpr, yvgO, wapA and nprE) were screened with a semi-rational approach and comparatively investigated their effects on the production of PGL. The signal peptide bpr directed efficient PGL secretory expression and increased PGL titer to 313.7 U mL(-1). By optimizing and applying strong promoter P43 and Shine-Dalgarno sequence, higher titer of 446.3 U mL(-1) PGL was achieved. Finally, the capacity of the recombinant B. subtilis WB43CB was evaluated with a fed-batch strategy in 3 L fermentor. The PGL titer reached 632.6 U mL(-1) with a productivity of 17.6 U mL(-1) h(-1), which was the highest secretory production of PGL by the B. subtilis system. The recombinant B. subtilis strain WB43CB constructed in the present work has great potential in production of alkaline PGL.
authors: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors: Junjiao
authors: J
authors: Zhang
authors: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China. Electronic address: zkang@jiangnan.edu.cn.
authors: Zhen
authors: Z
authors: Kang
authors: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors: Zhenmin
authors: Z
authors: Ling
authors: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors: Wenlong
authors: W
authors: Cao
authors: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors: Long
authors: L
authors: Liu
authors: School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
authors: Miao
authors: M
authors: Wang
authors: School of Biotechnology, Jiangnan University, Wuxi 214122, China; The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China. Electronic address: gcdu@jiangnan.edu.cn.
authors: Guocheng
authors: G
authors: Du
authors: School of Biotechnology, Jiangnan University, Wuxi 214122, China; National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China. Electronic address: jchen@jiangnan.edu.cn.
authors: Jian
authors: J
authors: Chen
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D019149:Bioreactors; D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D016297:Mutagenesis, Site-Directed; D009841:Oligonucleotides / Q000235:genetics; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011096:Polygalacturonase / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000096:biosynthesis*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D013997:Time Factors
chemical_list: D003470:Culture Media; D009841:Oligonucleotides; D010455:Peptides; D011994:Recombinant Proteins; D011096:Polygalacturonase; D011133:Polysaccharide-Lyases; C023911:pectate lyase
keywords: Alkaline polygalacturonate lyase; Bacillus subtilis; Signal peptide; bpr; promoter P43