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Effects of His-tag on Catalytic Activity and Enantioselectivity of Recombinant Transaminases

  • Lijun Meng
  • Yayun Liu
  • Xinjian Yin
  • Haisheng Zhou
  • Jianping Wu
  • Mianbin Wu
  • Lirong YangEmail author
Article
  • 95 Downloads

Abstract

Recombinant proteins were often expressed with His-tag to simplify the purification process. Among them, transaminase was mostly expressed with fusion tags and widely used in the production of numerous amino moieties. However, the existence of the His-tag has been reported to affect various properties of different recombinant enzymes, while the effect on transaminase was rarely studied. In this paper, we investigated the effect of His-tag on transaminase based on the various activities of 4-aminobutyrate-2-oxoglutarate transaminase (GabT) when it was expressed in vector pETDuet-1. We found that His-tag did not affect the enantioselectivity, but decreased the catalytic activity to different extents according to its existence and location. Native GabT maintained the highest catalytic activity; GabT with C-terminal His-tag showed slightly lower activity than native GabT but about 2.2-fold higher than GabT with N-terminal His-tag. Besides, other fusion tags like T7-tag and S-tag inserted between N-His-tag and GabT can relieve the decreasing effect of His-tag on GabT activity. Furthermore, whole cell catalytic activity of several transaminases was improved by deleting the N-terminal His-tag. This study provided a strategy for the efficient expression of recombinant transaminase with improved catalytic activity and might attract attention to the effect of His-tag on other enzymatic properties.

Keywords

His-tag Transaminase Catalytic activity Protein expression l-Phosphinothricin 

Notes

Funding information

This work was supported by the National Natural Science Foundation of China (No. 21476199).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12010_2019_3117_MOESM1_ESM.doc (109 kb)
ESM 1 (DOC 109 kb)

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Authors and Affiliations

  1. 1.Institute of Biological Engineering, College of Chemical and Biological EngineeringZhejiang UniversityHangzhouChina

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