title: Expression levels of chaperones influence biotransformation activity of recombinant Escherichia coli expressing Micrococcus luteus alcohol dehydrogenase and Pseudomonas putida Baeyer-Villiger monooxygenase.
abstract: We demonstrated for the first time that the archaeal chaperones (i.e., γ-prefoldin and thermosome) can stabilize enzyme activity in vivo. Ricinoleic acid biotransformation activity of recombinant Escherichia coli expressing Micrococcus luteus alcohol dehydrogenase and the Pseudomonas putida KT2440 Baeyer-Villiger monooxygenase improved significantly with co-expression of γ-prefoldin or recombinant themosome originating from the deep-sea hyperthermophile archaea Methanocaldococcus jannaschii. Furthermore, the degree of enhanced activity was dependent on the expression levels of the chaperones. For example, whole-cell biotransformation activity was highest at 12 µmol/g dry cells/min when γ-prefoldin expression level was approximately 46% of the theoretical maximum. This value was approximately two-fold greater than that in E. coli, where the γ-prefoldin expression level was zero or set to the theoretical maximum. Therefore, it was assumed that the expression levels of chaperones must be optimized to achieve maximum biotransformation activity in whole-cell biocatalysts.
authors.affiliation: Department of Food Science & Engineering, Ewha Womans University, Seoul, 120-750, Republic of Korea.
authors.forename: A-Hyong
authors.initials: AH
authors.lastname: Baek
authors.forename: Eun-Yeong
authors.initials: EY
authors.lastname: Jeon
authors.forename: Sun-Mee
authors.initials: SM
authors.lastname: Lee
authors.forename: Jin-Byung
authors.initials: JB
authors.lastname: Park
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D001711:Biotransformation; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D063747:Methanocaldococcus / Q000235:genetics* / Q000378:metabolism; D016982:Micrococcus luteus / Q000201:enzymology* / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012277:Ricinoleic Acids / Q000378:metabolism
chemical_list: D018832:Molecular Chaperones; D011994:Recombinant Proteins; D012277:Ricinoleic Acids; C112806:prefoldin; D006899:Mixed Function Oxygenases; D000426:Alcohol Dehydrogenase; C030521:ricinoleic acid
keywords: Baeyer-Villiger monooxygenase; Escherichia coli; alcohol dehydrogenase; chaperones; ricinoleic acid; thermosome; whole-cell biocatalysis; γ-prefoldin