title: Enzyme fusion for whole-cell biotransformation of long-chain sec-alcohols into esters.
abstract: Enzyme fusion was investigated as a strategy to improve productivity of a two-step whole-cell biocatalysis. The biotransformation of long-chain sec-alcohols into esters by an alcohol dehydrogenase (ADH) and Baeyer-Villiger monooxygenases (BVMOs) was used as the model reaction. The recombinant Escherichia coli, expressing the fusion enzymes between the ADH of Micrococcus luteus NCTC2665 and the BVMO of Pseudomonas putida KT2440 or Rhodococcus jostii RHA1, showed significantly greater bioconversion activity with long-chain sec-alcohols (e.g., 12-hydroxyoctadec-9-enoic acid (1a), 13-hydroxyoctadec-9-enoic acid (2a), 14-hydroxyicos-11-enoic acid (4a)) when compared to the recombinant E. coli expressing the ADH and BVMOs independently. For instance, activity of the recombinant E. coli expressing the ADH-Gly-BVMO, in which glycine-rich peptide was used as the linker, with 1a was increased up to 22 μmol g dry cells(-1) min(-1). This value is over 40 % greater than the recombinant E. coli expressing the ADH and BVMO independently. The substantial improvement appeared to be driven by an increase in the functional expression of the BVMOs and/or an increase in mass transport efficiency by localizing two active sites in close proximity.
authors: Department of Food Science & Engineering, Ewha Womans University, Seoul, 120-750, Republic of Korea.
authors: Eun-Yeong
authors: EY
authors: Jeon
authors: A-Hyong
authors: AH
authors: Baek
authors: Uwe T
authors: UT
authors: Bornscheuer
authors: Jin-Byung
authors: JB
authors: Park
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D000438:Alcohols / Q000378:metabolism*; D001711:Biotransformation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D004952:Esters / Q000378:metabolism*; D015870:Gene Expression; D016982:Micrococcus luteus / Q000201:enzymology / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012240:Rhodococcus / Q000201:enzymology / Q000235:genetics
chemical_list: D000438:Alcohols; D004952:Esters; D011993:Recombinant Fusion Proteins; D006899:Mixed Function Oxygenases; D000426:Alcohol Dehydrogenase