Improving the thermostability and catalytic efficiency of glucose oxidase from Aspergillus niger by molecular evolution.


Abstract

Glucose oxidase (Gox) has many applications in numerous industries. However, thermal instability is a major drawback that prevents its broader use. Here, Gox from Aspergillus niger (GoxA) was selected for laboratory evolution for purposes of enhancing thermostability and catalytic efficiency through random and rational mutagenesis. The most active mutant, M4, accumulated six amino acid substitutions. The T50 of M4, the temperature corresponding to a 50% loss of maximal enzyme activity, increased by 7.5 °C and thermal inactivation half-lives (t1/2) at 60 °C and 70 °C increased 8.4-fold and 5.6-fold, respectively, compared to wild-type GoxA. Concomitantly, M4 demonstrated a 1.86-fold increase in kcat, resulting in a 1.78-fold increase in catalytic efficiency. Molecular dynamics simulation revealed diverse mechanisms underlying the effects of each mutation on thermostability and catalytic efficiency. These results suggest that key properties of glucose oxidase can be modified in vitro by laboratory evolution, which may have remarkable economic importance.

Submission Details

ID: MntBGgaB

Submitter: Shu-Ching Ou

Submission Date: Jan. 25, 2019, 4:13 p.m.

Version: 1

Publication Details
Tu T;Wang Y;Huang H;Wang Y;Jiang X;Wang Z;Yao B;Luo H,Food Chem (2019) Improving the thermostability and catalytic efficiency of glucose oxidase from Aspergillus niger by molecular evolution. PMID:30658743
Additional Information

Data in Table 1 is not included as it mostly includes values from other studies.

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
3QVP 2011-06-01 1.2 Crystal structure of glucose oxidase for space group C2221 at 1.2 A resolution
3QVR 2011-06-01 1.3 Crystal structure of glucose oxidase for space group P3121 at 1.3 A resolution.
5NIW 2017-11-15 1.8 Glucose oxydase mutant A2
5NIT 2017-11-15 1.87 Glucose oxidase mutant A2
1CF3 1999-03-26 1.9 GLUCOSE OXIDASE FROM APERGILLUS NIGER
1GAL 1993-10-31 2.3 CRYSTAL STRUCTURE OF GLUCOSE OXIDASE FROM ASPERGILLUS NIGER: REFINED AT 2.3 ANGSTROMS RESOLUTION

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Glucose oxidase P13006 GOX_ASPNG