TY - JOUR
T1 - Enzyme fusion for whole-cell biotransformation of long-chain sec-alcohols into esters
AU - Jeon, Eun Yeong
AU - Baek, A. Hyong
AU - Bornscheuer, Uwe T.
AU - Park, Jin Byung
N1 - Funding Information:
This study was supported by the Marine Biomaterials Research Center grant from Marine Biotechnology Program funded by the Ministry of Oceans and Fisheries, Korea.
Publisher Copyright:
© 2015, Springer-Verlag Berlin Heidelberg.
PY - 2015/8/22
Y1 - 2015/8/22
N2 - 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.
AB - 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.
KW - Alcohol dehydrogenase
KW - Baeyer–Villiger monooxygenase
KW - Enzyme fusion
KW - Whole-cell biocatalysis
UR - http://www.scopus.com/inward/record.url?scp=84937639365&partnerID=8YFLogxK
U2 - 10.1007/s00253-015-6392-9
DO - 10.1007/s00253-015-6392-9
M3 - Article
C2 - 25636834
AN - SCOPUS:84937639365
SN - 0175-7598
VL - 99
SP - 6267
EP - 6275
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
IS - 15
ER -