title: Increased production of alkaline polygalacturonate lyase in the recombinant Pichia pastoris by controlling cell concentration during continuous culture.
abstract: Recombinant alkaline polygalacturonate lyase (PGL) production by recombinant Pichia pastoris GS115 was selected as a model to study as a continuous culture strategy for enhancing heterologous protein production based on controlling methanol feeding (CCCM culture) or on dual carbon source feeding (CCCD culture). Using the CCCM process with a dry cell weight of 75 g/L regulated by controlling methanol concentration in the induction media, the final PGL activity was 441.9 U/mL. The PGL productivity (Q(v)) and the average specific enzyme production rate (Q(x)) were 4.65 U mL(-1)h(-1) and 84.5 U g(-1)h(-1), an increase of 42.1% and 191.2%, respectively, over what was achieved with traditional fed-batch culture with high cell density. The control strategies also reduced proteolytic degradation by 84.1% in the fermentation broth and increased cell viability by 12.2%.
authors.affiliation: National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
authors.forename: Huilin
authors.initials: H
authors.lastname: Wang
authors.forename: Jianghua
authors.initials: J
authors.lastname: Li
authors.forename: Long
authors.initials: L
authors.lastname: Liu
authors.forename: Xiaoman
authors.initials: X
authors.lastname: Li
authors.forename: Dongxu
authors.initials: D
authors.lastname: Jia
authors.forename: Guocheng
authors.initials: G
authors.lastname: Du
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.forename: Jiangning
authors.initials: J
authors.lastname: Song
mesh_terms: D018533:Biomass; D018929:Cell Culture Techniques; D003470:Culture Media; D006863:Hydrogen-Ion Concentration; D010843:Pichia / Q000235:genetics*; D011133:Polysaccharide-Lyases / Q000096:biosynthesis*; D011995:Recombination, Genetic
chemical_list: D003470:Culture Media; D011133:Polysaccharide-Lyases; C023911:pectate lyase