title: Secretory production of recombinant protein by a high cell density culture of a protease negative mutant Escherichia coli strain.
abstract: Several protease negative mutant strains including HM114, HM126, and HM130 as well as their parent strain KS272 were compared for their growth and secretory production of a model fusion protein, protein A-beta-lactamase. HM114, a strain deficient in two cell envelope proteases, grew slightly faster and produced more fusion protein than the other strains deficient in more proteases. HM114 was grown to a cell dry weight of 47.86 g/L in 29 h using pH-stat, fed-batch cultivation. The beta-lactamase activity was 11.25 x 10(4) U/L, which was 30% higher than that obtained with its parent strain KS272. Up to 96% of protein A-beta-lactamase fusion protein could be recovered by a simple cold osmotic shock method. The specific beta-lactamase activity obtained with HM114 after fractionation was 4.5 times higher than that obtained with KS272.
authors: Department of Chemical Engineering and BioProcess Engineering Research Center, Korea Advanced Institute of Science and Technology, 373-1 Kusong-dong, Yusong-gu, Taejon 305-701, Korea.
authors: S J
authors: SJ
authors: Park
authors: G
authors: G
authors: Georgiou
authors: S Y
authors: SY
authors: Lee
mesh_terms: D010450:Endopeptidases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development*; D005285:Fermentation; D009154:Mutation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D010450:Endopeptidases; D001618:beta-Lactamases