title: Overproduction of a truncated poly (vinyl alcohol) dehydrogenase in recombinant Pichia pastoris by low-temperature induction strategy and related mechanism analysis.
abstract: Based on the N-terminal sequencing of poly (vinyl alcohol) dehydrogenase (PVADH), a 1,644-bp gene encoding a truncated PVADH (tPVADH) was amplified using the synthetic gene (GenBank accession No. JQ235753) as a template, and was further transformed into Pichia pastoris GS115 with the vector pPIC9K. The maximal tPVADH activity reached 546 U/mL in shake flask. The influence of methanol concentration and induction temperature on tPVADH production was further investigated in 3-L bioreactor. When the methanol concentration and induction temperature were controlled at 15 g/L and 22 °C, respectively, the maximal tPVADH activity reached 8,464 U/mL, which was nearly 10 times that of mature PVADH expressed under the same condition and was the highest level ever reported. The reason responsible for the significant improvement of tPVADH production at low induction temperature was explored in terms of cell viability, extracellular proteases activity, and alcohol oxidase activity.
authors.affiliation: Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
authors.forename: Dongxu
authors.initials: D
authors.lastname: Jia
authors.forename: Long
authors.initials: L
authors.lastname: Liu
authors.forename: Huilin
authors.initials: H
authors.lastname: Wang
authors.forename: Dongxu
authors.initials: D
authors.lastname: Zhang
authors.forename: Jianghua
authors.initials: J
authors.lastname: Li
authors.forename: Guocheng
authors.initials: G
authors.lastname: Du
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
mesh_terms: D000429:Alcohol Oxidoreductases / Q000096:biosynthesis* / Q000737:chemistry; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D003001:Cloning, Molecular; D003080:Cold Temperature; D017931:DNA Primers; D005285:Fermentation; D007218:Industrial Microbiology / Q000379:methods*; D010843:Pichia / Q000378:metabolism*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D013997:Time Factors
chemical_list: D017931:DNA Primers; D000429:Alcohol Oxidoreductases; C048312:polyvinyl-alcohol dehydrogenase (acceptor); C019643:alcohol oxidase