PMID- 36439831
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20221129
IS  - 1664-302X (Print)
IS  - 1664-302X (Electronic)
IS  - 1664-302X (Linking)
VI  - 13
DP  - 2022
TI  - Directed evolution engineering to improve activity of glucose dehydrogenase by 
      increasing pocket hydrophobicity.
PG  - 1044226
LID - 10.3389/fmicb.2022.1044226 [doi]
LID - 1044226
AB  - Glucose dehydrogenase (GDH) is a NAD(P)(+) dependent oxidoreductase, which is 
      useful in glucose determination kits, glucose biosensors, cofactor regeneration, 
      and biofuel cells. However, the low efficiency of the catalysis hinders the use 
      of GDH in industrial applications. In this study, an analysis of interactions 
      between eight GDH mutants and NADP(+) is powered by AlphaFold2 and Discovery 
      Studio 3.0. The docking results showed that more hydrogen bonds formed between 
      mutants, such as P45A and NADP(+), which indicated that these mutants had the 
      potential for high catalytic efficiency. Subsequently, we verified all the 
      mutants by site-directed mutagenesis. It was notable that the enzyme activity of 
      mutant P45A was 1829 U/mg, an improvement of 28-fold compared to wild-type GDH. 
      We predicted the hydrophobicity of the protein-ligand complexes, which was 
      confirmed by an 8-anilino-1-naphthalenesulphonic acid fluorescent probe. The 
      following order of increasing hydrophobicity index was deduced: 
      GDH < N46E < F155Y < P45A, which suggested that the enzyme activity of GDH is 
      positively related to its pocket hydrophobicity. Furthermore, P45A still showed 
      better catalytic ability in organic solvents, reaching 692 U/mg in 10% 
      isopropanol, which was 19-fold that of the wild-type GDH. However, its substrate 
      affinity was affected by organic solvents. This study provides a good theoretical 
      foundation for further improving the catalytic efficiency of GDH.
CI  - Copyright (c) 2022 Hua, Qianqian, Jianfeng, Yinbiao, Shengyu, Weishi, Yang and 
      Yupeng.
FAU - Hua, Li
AU  - Hua L
AD  - School of Life Sciences, Institute of Microbial Engineering, Henan University, 
      Kaifeng, China.
AD  - Engineering Research Center for Applied Microbiology of Henan Province, Henan 
      University, Kaifeng, China.
FAU - Qianqian, Bao
AU  - Qianqian B
AD  - School of Life Sciences, Institute of Microbial Engineering, Henan University, 
      Kaifeng, China.
AD  - Engineering Research Center for Applied Microbiology of Henan Province, Henan 
      University, Kaifeng, China.
FAU - Jianfeng, Zhao
AU  - Jianfeng Z
AD  - School of Life Sciences, Institute of Microbial Engineering, Henan University, 
      Kaifeng, China.
AD  - Engineering Research Center for Applied Microbiology of Henan Province, Henan 
      University, Kaifeng, China.
FAU - Yinbiao, Xu
AU  - Yinbiao X
AD  - School of Life Sciences, Institute of Microbial Engineering, Henan University, 
      Kaifeng, China.
AD  - Engineering Research Center for Applied Microbiology of Henan Province, Henan 
      University, Kaifeng, China.
FAU - Shengyu, Yang
AU  - Shengyu Y
AD  - School of Life Sciences, Institute of Microbial Engineering, Henan University, 
      Kaifeng, China.
AD  - Engineering Research Center for Applied Microbiology of Henan Province, Henan 
      University, Kaifeng, China.
FAU - Weishi, Xue
AU  - Weishi X
AD  - School of Life Sciences, Institute of Microbial Engineering, Henan University, 
      Kaifeng, China.
AD  - Engineering Research Center for Applied Microbiology of Henan Province, Henan 
      University, Kaifeng, China.
FAU - Yang, Sun
AU  - Yang S
AD  - School of Life Sciences, Institute of Microbial Engineering, Henan University, 
      Kaifeng, China.
AD  - Engineering Research Center for Applied Microbiology of Henan Province, Henan 
      University, Kaifeng, China.
FAU - Yupeng, Liu
AU  - Yupeng L
AD  - School of Life Sciences, Institute of Microbial Engineering, Henan University, 
      Kaifeng, China.
AD  - Engineering Research Center for Applied Microbiology of Henan Province, Henan 
      University, Kaifeng, China.
LA  - eng
PT  - Journal Article
DEP - 20221109
PL  - Switzerland
TA  - Front Microbiol
JT  - Frontiers in microbiology
JID - 101548977
PMC - PMC9681798
OTO - NOTNLM
OT  - AlphaFold2
OT  - glucose dehydrogenase
OT  - hydrophobicity
OT  - organic solvents
OT  - site-directed mutagenesis
COIS- The authors declare that the research was conducted in the absence of any 
      commercial or financial relationships that could be construed as a potential 
      conflict of interest.
EDAT- 2022/11/29 06:00
MHDA- 2022/11/29 06:01
PMCR- 2022/11/09
CRDT- 2022/11/28 04:44
PHST- 2022/09/14 00:00 [received]
PHST- 2022/10/17 00:00 [accepted]
PHST- 2022/11/28 04:44 [entrez]
PHST- 2022/11/29 06:00 [pubmed]
PHST- 2022/11/29 06:01 [medline]
PHST- 2022/11/09 00:00 [pmc-release]
AID - 10.3389/fmicb.2022.1044226 [doi]
PST - epublish
SO  - Front Microbiol. 2022 Nov 9;13:1044226. doi: 10.3389/fmicb.2022.1044226. 
      eCollection 2022.