title: Bacteriocin release protein-mediated secretory expression of recombinant chalcone synthase in Escherichia coli.
abstract: Flavonoids are secondary metabolites synthesized by plants shown to exhibit health benefits such as anti-inflammatory, antioxidant, and anti-tumor effects. Thus, due to the importance of this compound, several enzymes involved in the flavonoid pathway have been cloned and characterized in Escherichia coli. However, the formation of inclusion bodies has become a major disadvantage of this approach. As an alternative, chalcone synthase from Physcomitrella patens was secreted into the medium using a bacteriocin release protein expression vector. Secretion of P. patens chalcone synthase into the culture media was achieved by co-expression with a psW1 plasmid encoding bacteriocin release protein in E. coli Tuner (DE3) plysS. The optimized conditions, which include the incubation of cells for 20 h with 40 ng/ml mitomycin C at OD(600) induction time of 0.5 was found to be the best condition for chalcone synthase secretion.
authors.affiliation: Faculty of Biotechnology and Biomolecular Sciences, Enzyme and Microbial Technology Research Group, Universiti Putra Malaysia, Serdang, Malaysia. milounited_86@yahoo.com
authors.forename: Iffah Izzati
authors.initials: II
authors.lastname: Zakaria
authors.forename: Raja Noor Zaliha Raja Abdul
authors.initials: RN
authors.lastname: Rahman
authors.forename: Abu Bakar
authors.initials: AB
authors.lastname: Salleh
authors.forename: Mahiran
authors.initials: M
authors.lastname: Basri
mesh_terms: D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001430:Bacteriocins / Q000235:genetics*; D001709:Biotechnology / Q000379:methods*; D019068:Bryopsida / Q000737:chemistry / Q000201:enzymology*; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005110:Extracellular Space / Q000201:enzymology; D015870:Gene Expression / Q000187:drug effects; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D016685:Mitomycin / Q000494:pharmacology; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D053719:Tandem Mass Spectrometry; D014169:Transformation, Bacterial
chemical_list: D001430:Bacteriocins; D010940:Plant Proteins; D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside; D016685:Mitomycin; D000217:Acyltransferases; C020141:flavanone synthetase