PMID- 32324965
OWN - NLM
STAT- MEDLINE
DCOM- 20210707
LR  - 20210707
IS  - 1439-7633 (Electronic)
IS  - 1439-4227 (Linking)
VI  - 21
IP  - 18
DP  - 2020 Sep 14
TI  - Evolution of Glucose Dehydrogenase for Cofactor Regeneration in Bioredox 
      Processes with Denaturing Agents.
PG  - 2680-2688
LID - 10.1002/cbic.202000196 [doi]
AB  - Glucose dehydrogenase (GDH) is a general tool for driving nicotinamide (NAD(P)H) 
      regeneration in synthetic biochemistry. An increasing number of synthetic 
      bioreactions are carried out in media containing high amounts of organic 
      cosolvents or hydrophobic substrates/products, which often denature native 
      enzymes, including those for cofactor regeneration. In this work, we attempted to 
      improve the chemical stability of Bacillus megaterium GDH (BmGDH(M0) ) in the 
      presence of large amounts of 1-phenylethanol by directed evolution. Among the 
      resulting mutants, BmGDH(M6) (Q252L/E170K/S100P/K166R/V72I/K137R) exhibited a 
      9.2-fold increase in tolerance against 10 % (v/v) 1-phenylethanol. Moreover, 
      BmGDH(M6) was also more stable than BmGDH(M0) when exposed to hydrophobic and 
      enzyme-inactivating compounds such as acetophenone, ethyl 2-oxo-4-phenylbutyrate, 
      and ethyl (R)-2-hydroxy-4-phenylbutyrate. Coupled with a Candida glabrata 
      carbonyl reductase, BmGDH(M6) was successfully used for the asymmetric reduction 
      of deactivating ethyl 2-oxo-4-phenylbutyrate with total turnover number of 1800 
      for the nicotinamide cofactor, thus making it attractive for commercial 
      application. Overall, the evolution of chemically robust GDH facilitates its 
      wider use as a general tool for NAD(P)H regeneration in biocatalysis.
CI  - (c) 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
FAU - Qian, Wen-Zhuo
AU  - Qian WZ
AD  - State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation 
      Center for Biomanufacturing, East China University of Science and Technology, 130 
      Meilong Road, Shanghai, 200237, P. R. China.
FAU - Ou, Ling
AU  - Ou L
AD  - State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation 
      Center for Biomanufacturing, East China University of Science and Technology, 130 
      Meilong Road, Shanghai, 200237, P. R. China.
FAU - Li, Chun-Xiu
AU  - Li CX
AD  - State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation 
      Center for Biomanufacturing, East China University of Science and Technology, 130 
      Meilong Road, Shanghai, 200237, P. R. China.
FAU - Pan, Jiang
AU  - Pan J
AD  - State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation 
      Center for Biomanufacturing, East China University of Science and Technology, 130 
      Meilong Road, Shanghai, 200237, P. R. China.
FAU - Xu, Jian-He
AU  - Xu JH
AD  - State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation 
      Center for Biomanufacturing, East China University of Science and Technology, 130 
      Meilong Road, Shanghai, 200237, P. R. China.
FAU - Chen, Qi
AU  - Chen Q
AD  - State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation 
      Center for Biomanufacturing, East China University of Science and Technology, 130 
      Meilong Road, Shanghai, 200237, P. R. China.
FAU - Zheng, Gao-Wei
AU  - Zheng GW
AUID- ORCID: 0000-0001-5918-6047
AD  - State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation 
      Center for Biomanufacturing, East China University of Science and Technology, 130 
      Meilong Road, Shanghai, 200237, P. R. China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200604
PL  - Germany
TA  - Chembiochem
JT  - Chembiochem : a European journal of chemical biology
JID - 100937360
RN  - 0 (Benzyl Alcohols)
RN  - 0 (Phenylbutyrates)
RN  - 0 (ethyl 2-oxo-4-phenylbutyrate)
RN  - 25X51I8RD4 (Niacinamide)
RN  - E6O895DQ52 (methylphenyl carbinol)
RN  - EC 1.1.1.47 (Glucose 1-Dehydrogenase)
SB  - IM
MH  - Bacillus megaterium/enzymology
MH  - Benzyl Alcohols/chemistry/metabolism
MH  - Glucose 1-Dehydrogenase/chemistry/genetics/*metabolism
MH  - Molecular Structure
MH  - Mutation
MH  - Niacinamide/chemistry/*metabolism
MH  - Oxidation-Reduction
MH  - Phenylbutyrates/chemistry/metabolism
OTO - NOTNLM
OT  - biocatalysis
OT  - chemical stability
OT  - cofactor regeneration
OT  - directed evolution
OT  - glucose dehydrogenase 
EDAT- 2020/04/24 06:00
MHDA- 2021/07/08 06:00
CRDT- 2020/04/24 06:00
PHST- 2020/03/29 00:00 [received]
PHST- 2020/04/20 00:00 [revised]
PHST- 2020/04/24 06:00 [pubmed]
PHST- 2021/07/08 06:00 [medline]
PHST- 2020/04/24 06:00 [entrez]
AID - 10.1002/cbic.202000196 [doi]
PST - ppublish
SO  - Chembiochem. 2020 Sep 14;21(18):2680-2688. doi: 10.1002/cbic.202000196. Epub 2020 
      Jun 4.