title: Efficient expression of an alkaline pectate lyase gene from Bacillus subtilis and the characterization of the recombinant protein.
abstract: The gene encoding a novel alkaline pectate lyase (Apel) from Bacillus subtilis was cloned and expressed in B. subtilis WB600. Apel contained an ORF of 1,260 bp, encoding a signal peptide of 21 amino acids and a mature protein of 399 amino acids with a calculated molecular mass of 45497.9 Da. The mature Apel was structurally related to the enzymes in the polysaccharide lyase family 1. After purification, the recombinant Apel had a specific activity of 445 U mg(-1). The enzyme was optimally active at 50°C and pH 9.
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin, 300457, People's Republic of China. lyh@tust.edu.cn
authors.forename: Yihan
authors.initials: Y
authors.lastname: Liu
authors.forename: Guanqun
authors.initials: G
authors.lastname: Chen
authors.forename: Jianling
authors.initials: J
authors.lastname: Wang
authors.forename: Yujie
authors.initials: Y
authors.lastname: Hao
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
authors.forename: Bo
authors.initials: B
authors.lastname: Hu
authors.forename: Fuping
authors.initials: F
authors.lastname: Lu
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004795:Enzyme Stability; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013696:Temperature
chemical_list: D004269:DNA, Bacterial; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D011133:Polysaccharide-Lyases; C023911:pectate lyase