title: Alleviation of proteolytic sensitivity to enhance recombinant lipase production in Escherichia coli.
abstract: Two amino acids, Leu149 and Val223, were identified as proteolytically sensitive when Pseudozyma antarctica lipase (PalB) was heterologously expressed in Escherichia coli. The functional expression was enhanced using the double mutant for cultivation. However, the recombinant protein production was still limited by PalB misfolding, which was resolved by DsbA coexpression.
authors: Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
authors: Niju
authors: N
authors: Narayanan
authors: C Perry
authors: CP
authors: Chou
mesh_terms: D019943:Amino Acid Substitution; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D007218:Industrial Microbiology / Q000379:methods; D007930:Leucine / Q000235:genetics; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014587:Ustilaginales / Q000201:enzymology / Q000235:genetics; D014633:Valine / Q000235:genetics
chemical_list: D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D008049:Lipase; D010447:Peptide Hydrolases; D019704:Protein Disulfide-Isomerases; C526648:dsbA protein, E coli; D007930:Leucine; D014633:Valine