PMID- 27363470
OWN - NLM
STAT- MEDLINE
DCOM- 20170522
LR  - 20170522
IS  - 1738-8872 (Electronic)
IS  - 1017-7825 (Linking)
VI  - 26
IP  - 10
DP  - 2016 Oct 28
TI  - Effects of N-/C-Terminal Extra Tags on the Optimal Reaction Conditions, Activity, 
      and Quaternary Structure of Bacillus thuringiensis Glucose 1-Dehydrogenase.
PG  - 1708-1716
LID - 10.4014/jmb.1603.03021 [doi]
AB  - Glucose dehydrogenase (GDH) is an oxidoreductase enzyme and is used as a 
      biocatalyst to regenerate NAD(P)H in reductase-mediated chiral synthesis 
      reactions. In this study, the glucose 1-dehydrogenase B gene (gdhB) was cloned 
      from Bacillus thuringiensis subsp. kurstaki, and wild-type (GDH-BT(WT)) and 
      His-tagged (GDH-BT(N-His), GDH-BT(C-His)) enzymes were produced in Escherichia 
      coli BL21 (DE3). All enzymes were produced in the soluble forms from E. coli. 
      GDH-BT(WT) and GDH-BT(N-His) showed high specific enzymatic activities of 6.6 
      U/mg and 5.5 U/mg, respectively, whereas GDH-BT(C-His) showed a very low specific 
      enzymatic activity of 0.020 U/mg. These results suggest that the intact 
      C-terminal carboxyl group is important for GDH-BT activity. GDH-BT(WT) was stable 
      up to 65 degrees C, whereas GDH-BT(N-His) and GDH-BT(C-His) were stable up to 45 degrees C. Gel 
      permeation chromatography showed that GDH-BT(WT) is a dimer, whereas 
      GDH-BT(N-His) and GDH-BT(C-His) are monomeric. These results suggest that the 
      intact N- and C-termini are required for GDH-BT to maintain thermostability and 
      to form its dimer structure. The homology model of the GDH-BT(WT) single subunit 
      was constructed based on the crystal structure of Bacillus megaterium GDH (PDB ID 
      3AY6), showing that GDH-BT(WT) has a Rossmann fold structure with its N- and 
      C-termini located on the subunit surface, which suggests that His-tagging 
      affected the native dimer structure. GDH-BT(WT) and GDH-BT(N-His) regenerated 
      NADPH in a yeast reductase-mediated chiral synthesis reaction, suggesting that 
      these enzymes can be used as catalysts in fine-chemical and pharmaceutical 
      industries.
FAU - Hyun, Jeongwoo
AU  - Hyun J
AD  - Division of Biotechnology, The Catholic University of Korea, Bucheon 14662, 
      Republic of Korea.
FAU - Abigail, Maria
AU  - Abigail M
AD  - Division of Biotechnology, The Catholic University of Korea, Bucheon 14662, 
      Republic of Korea.
AD  - Faculty of Biotechnology, Atma Jaya Catholic University of Indonesia, Jakarta 
      12930, Indonesia.
FAU - Choo, Jin Woo
AU  - Choo JW
AD  - Division of Biotechnology, The Catholic University of Korea, Bucheon 14662, 
      Republic of Korea.
FAU - Ryu, Jin
AU  - Ryu J
AD  - Division of Biotechnology, The Catholic University of Korea, Bucheon 14662, 
      Republic of Korea.
FAU - Kim, Hyung Kwoun
AU  - Kim HK
AD  - Division of Biotechnology, The Catholic University of Korea, Bucheon 14662, 
      Republic of Korea.
LA  - eng
PT  - Journal Article
PL  - Korea (South)
TA  - J Microbiol Biotechnol
JT  - Journal of microbiology and biotechnology
JID - 9431852
RN  - 0 (Recombinant Proteins)
RN  - 53-59-8 (NADP)
RN  - EC 1.1.1.47 (Glucose 1-Dehydrogenase)
SB  - IM
MH  - Bacillus thuringiensis/enzymology/*genetics
MH  - Cloning, Molecular
MH  - Enzyme Stability
MH  - Escherichia coli/genetics
MH  - Glucose 1-Dehydrogenase/*chemistry/genetics/*metabolism
MH  - Kinetics
MH  - Models, Molecular
MH  - NADP/metabolism
MH  - Recombinant Proteins/*chemistry/genetics/*metabolism
MH  - Substrate Specificity
OTO - NOTNLM
OT  - Bacillus thuringiensis
OT  - Glucose dehydrogenase
OT  - His-tag
OT  - NADPH regeneration
OT  - homology model
EDAT- 2016/10/27 06:00
MHDA- 2017/05/23 06:00
CRDT- 2016/07/02 06:00
PHST- 2016/10/27 06:00 [pubmed]
PHST- 2017/05/23 06:00 [medline]
PHST- 2016/07/02 06:00 [entrez]
AID - 10.4014/jmb.1603.03021 [pii]
AID - 10.4014/jmb.1603.03021 [doi]
PST - ppublish
SO  - J Microbiol Biotechnol. 2016 Oct 28;26(10):1708-1716. doi: 
      10.4014/jmb.1603.03021.