title: Construction and application of recombinant strain for the production of an alkaline protease from Bacillus licheniformis.
abstract: The alkaline protease gene, Apr, from Bacillus licheniformis 2709 was cloned into an expression vector pET - 28b (+), to yield the recombinant plasmid pET-28b (+) - Apr. The pET-28b (+) - Apr was expressed in a high expression strain E. coli BL21. The amino acid sequence deduced from the DNA sequence analysis revealed a 98% identity to that of Bacillus licheniformis 2709. Sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE) was used to access the protein expression. SDS-PAGE analysis indicated a protein of Mr of 38.8 kDa. The medium components and condition of incubation were optimized for the growth state of a recombinant strain. The optimal composition of production medium was composed of glucose 8 g/L, peptone 8 g/L and salt solution 10 mL. The samples were incubated on a rotary shaker of 180 r/min at 37°C for 24 h.
authors.affiliation: Laboratory of Nutrition and Functional Food, Jilin University, Changchun 130062, PR China.
authors.forename: Songyi
authors.initials: S
authors.lastname: Lin
authors.affiliation: Laboratory of Nutrition and Functional Food, Jilin University, Changchun 130062, PR China.
authors.forename: Meishuo
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Laboratory of Nutrition and Functional Food, Jilin University, Changchun 130062, PR China. Electronic address: liujingbo_jlu@163.com.
authors.forename: Jingbo
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Food, Nutrition and Packaging Sciences, Clemson University, Clemson, SC 29634, USA.
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Jones
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics*; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D055162:Biocatalysis / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010461:Peptones / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D017422:Sequence Analysis, DNA; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D010461:Peptones; D011994:Recombinant Proteins; D010450:Endopeptidases; C087822:alkaline protease; D005947:Glucose
keywords: Activity; Alkaline protease; Bacillus licheniformis; Optimization; Recombinant