title: Optimization of the production of Aspergillus niger α-glucosidase expressed in Pichia pastoris.
abstract: The α-glucosidase (AGL) from Aspergillus niger has been applied to produce isomaltooligosaccharides. In the present study, various factors which affect the yield of recombinant AGL, produced by engineered Pichia pastoris, were investigated. The expression level reached 5.5 U ml(-1) in bioreactor after optimization of parameters of initial induction cell density, induction temperature and methanol concentration. In addition, it was found that coexpression of protein disulfide isomerase (PDI) inhibited the growth of the engineered P. pastoris strains and had an adverse effect on the production of AGL, while codon optimization of native A. niger α-glucosidase encoding gene (aglu) resulted in a significant enhancement of enzyme production, which reached 10.1 U ml(-1). We believe that yield of AGL is increased by codon optimization as a result of enhanced translation efficiency as well as more stable mRNA secondary structure. In contrast, PDI coexpression under the control of alcohol oxidase promoter (PAOX1) seems to be less efficient in helping disulfide bond formation in AGL while probably induce unfolded protein response, which further leads to cell apoptosis and increased protein degradation.
authors.affiliation: School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, Wuxi 214122, China. liuxu880718@hotmail.com
authors.forename: Xu
authors.initials: X
authors.lastname: Liu
authors.forename: Dan
authors.initials: D
authors.lastname: Wu
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D001709:Biotechnology / Q000379:methods*; D003001:Cloning, Molecular; D003062:Codon / Q000378:metabolism; D005285:Fermentation; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D000520:alpha-Glucosidases / Q000235:genetics / Q000378:metabolism*
chemical_list: D003062:Codon; D011994:Recombinant Proteins; D000520:alpha-Glucosidases; D019704:Protein Disulfide-Isomerases