Enhancement of cell viability and alkaline polygalacturonate lyase production by sorbitol co-feeding with methanol in Pichia pastoris fermentation.
Alkaline polygalacturonate lyase (PGL) production by Pichia pastoris GS115 was used as a model to study the mechanism and strategy for enhancing heterologous protein production. In order to enhance cell viability and volumetric recombinant protein productivity, sorbitol, which had been confirmed to be a non-repressive carbon source, was added together with methanol during the induction phase. The resultant PGL activity was up to 1593 U mL(-1), which was enhanced 1.85-fold compared to the control (863 U mL(-1)) cultured with sorbitol added at a constant rate of 3.6 g h(-1)L(-1) after an induction period of 100 h. Further results revealed that an appropriate sorbitol co-feeding strategy not only decreased the cell mortality to 8.8% (the control is about 23.1%) in the end of fermentation, but also reduced the proteolytic degradation of PGL.
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122 Jiangsu, China.
Zhihao
Z
Wang
Yun
Y
Wang
Dongxu
D
Zhang
Jianghua
J
Li
Zhaozhe
Z
Hua
Guocheng
G
Du
Jian
J
Chen
D000429:Alcohol Oxidoreductases / Q000378:metabolism; D000468:Alkalies / Q000378:metabolism; D018533:Biomass; D005285:Fermentation / Q000187:drug effects*; D042541:Intracellular Space / Q000187:drug effects / Q000201:enzymology; D000432:Methanol / Q000494:pharmacology*; D050296:Microbial Viability / Q000187:drug effects*; D010447:Peptide Hydrolases / Q000378:metabolism; D010843:Pichia / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D011133:Polysaccharide-Lyases / Q000096:biosynthesis*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D013012:Sorbitol / Q000494:pharmacology*; D013696:Temperature; D013997:Time Factors
D000468:Alkalies; D013012:Sorbitol; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D010447:Peptide Hydrolases; D011133:Polysaccharide-Lyases; C023911:pectate lyase; D000432:Methanol