Enhancement of alkaline polygalacturonate lyase production in recombinant Pichia pastoris according to the ratio of methanol to cell concentration.
Polygalacturonate lyase (PGL) production by Pichia pastoris GS115 was used as a model to study the mechanism and strategy for enhancing heterologous protein production. It was found that the ratio of methanol to cell concentration had a significant influence on PGL production. In this study, an advanced glycerol exponential feeding strategy was developed for biomass accumulation in cell growth phase, by which cell concentration reached 140 g L(-1) after 19 h glycerol feeding. In subsequent production phase, a methanol feeding profile was proposed according to the optimal ratio of methanol to cell concentration at a range of 0.163-0.171 g g(-1), and PGL activity and productivity reached 430 U mL(-1) and 4.34 U mL(-1)h(-1), respectively. The strategy for enhancing PGL production by controlling the optimal ratio may provide an alternative approach to enhance heterologous protein production with P. pastoris.
Wang
Yun
Y
Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
Wang
Zhihao
Z
Du
Guocheng
G
Hua
Zhaozhe
Z
Liu
Liming
L
Li
Jianghua
J
Chen
Jian
J
D000468:Alkalies / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods*; D000432:Methanol / Q000378:metabolism*; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011133:Polysaccharide-Lyases / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism*
D000468:Alkalies; D011994:Recombinant Proteins; D011133:Polysaccharide-Lyases; C023911:pectate lyase; D000432:Methanol