title: Efficient secretion of lipase r27RCL in Pichia pastoris by enhancing the disulfide bond formation pathway in the endoplasmic reticulum.
abstract: The lipase r27RCL from Rhizopus chinensis CCTCC M201021 was heterologously expressed in Pichia pastoris GS115 by simultaneous co-expression with two secretion factors ERO1p and PDI involved in the endoplasmic reticulum (ER). Compared to the expression of the lipase alone (12,500 U/ml), co-expression with these two proteins resulted in the production of larger total quantities of enzymes. The largest increase was seen when the combined ERO1p/PDI system was co-expressed, resulting in approximately 30 % higher enzyme yields (16,200 U/ml) than in the absence of co-expressed secretion factors. The extracellular protein concentration of the recombinant strain Co XY RCL-5 reached 9.39 g/l in the 7-l fermentor. Simultaneously, the fermentation time was also shortened by about 8 h compared to that of the control. The substrate-specific consumption rate (Qs) and the product-specific production rate (Qp) were both investigated in this research. In conclusion, the space-time yield was improved by co-expression with ERO1p and PDI. This is a potential strategy for high level expression of other heterologous proteins in P. pastoris.
authors.affiliation: State Key Laboratory of Food Science and Technology, The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 lihu Avenue, Wuxi, 214122, Jiangsu, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Sha
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Yu
authors.forename: Meng
authors.initials: M
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
mesh_terms: D000818:Animals; D019149:Bioreactors; D004220:Disulfides / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005285:Fermentation; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D008049:Lipase / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D012233:Rhizopus / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D004220:Disulfides; D005656:Fungal Proteins; D006023:Glycoproteins; D008049:Lipase; D019704:Protein Disulfide-Isomerases