Computational Design of Multisubstrate Enzyme Specificity


The engineering of multi-substrate enzyme specificity is highly desirable to foster the application of biocatalysis in industry. Here, we develop a multistate computational protein design methodology called multi-chemical state analysis (MCSA) that can optimize enzyme sequences on large structural ensembles for productive binding of multiple target substrates. Using MCSA, we redesigned E. coli branched-chain amino acid aminotransferase to accept both α-ketoglutarate and the non-native substrate L-histidine. Screening of a designed combinatorial library comprising 32 mutants for enhanced L-histidine transamination activity yielded four variants displaying up to ≈200-fold improvements to kcat/KM. MCSA opens the door to the design of broad-specificity biocatalysts and multi-substrate enzymes displaying tailored specificity.

Submission Details

ID: DddCjJY34

Submitter: Antony St-Jacques

Submission Date: May 7, 2020, 7:42 a.m.

Version: 1

Publication Details
Antony D. St-Jacques, Marie-Ève C. Eyahpaise, Roberto A. Chica,ACS Catalysis (2019) Computational Design of Multisubstrate Enzyme Specificity
Additional Information

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 UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
300.0 A,B,C Branched-chain-amino-acid aminotransferase P0AB82 ILVE_ECO57
300.0 A,B,C Branched-chain-amino-acid aminotransferase P0AB81 ILVE_ECOL6
300.0 A,B,C Branched-chain-amino-acid aminotransferase P0AB80 ILVE_ECOLI
294.29999999999995 A,B,C Branched-chain-amino-acid aminotransferase P0A1A6 ILVE_SALTI
294.29999999999995 A,B,C Branched-chain-amino-acid aminotransferase P0A1A5 ILVE_SALTY