The engineering of decameric D-fructose-6-phosphate aldolase A by combinatorial modulation of inter- and intra-subunit interactions

Xiaohong Yang; Lian Wu; Aipeng Li; Lidan Ye; Jiahai Zhou; Hongwei Yu

doi:10.1039/D0CC02437F

The engineering of decameric d-fructose-6-phosphate aldolase A by combinatorial modulation of inter- and intra-subunit interactions†

Xiaohong Yang,‡^ab Lian Wu,‡^cd Aipeng Li,^a Lidan Ye,

*^ab Jiahai Zhou

*^c and Hongwei Yu

*^ab

Author affiliations

* Corresponding authors

^a Key Laboratory of Biomass Chemical Engineering (Education Ministry), College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China
E-mail: yuhongwei@zju.edu.cn, yelidan@zju.edu.cn

^b Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China

^c State Key Laboratory of Bioorganic and Natural Products Chemistry, Center for Excellence in Molecular Synthesis, Shanghai Institute of Organic Chemistry, University of Chinese Academy of Sciences, 345 Lingling Road, Shanghai 200032, China
E-mail: jiahai@sioc.ac.cn

^d The Key Laboratory of Synthetic Biology, and CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China

Abstract

The combinatorial modulation of inter- and intra-subunit interactions of decameric D-fructose-6-phosphate aldolase A (FSAA) generated a triple-site variant I31T/Q59T/I195Q FSAA with 27- to 278-fold improvement in activity towards target heteroaromatic aldehydes. X-ray crystallographic data and molecular dynamics simulations ascribed the enhanced activity to the pronounced flexibility of the interface region between subunits, the expanded substrate entrance and binding pocket, and enhanced proton transfer, unambiguously demonstrating the efficiency of this strategy for engineering multimeric enzymes.

This article is part of the themed collection: Chemical Communications HOT Articles

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DOI: https://doi.org/10.1039/D0CC02437F
Article type: Communication
Submitted: 04 Apr 2020
Accepted: 01 May 2020
First published: 07 May 2020

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Chem. Commun., 2020,56, 7561-7564

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The engineering of decameric D-fructose-6-phosphate aldolase A by combinatorial modulation of inter- and intra-subunit interactions

X. Yang, L. Wu, A. Li, L. Ye, J. Zhou and H. Yu, Chem. Commun., 2020, 56, 7561 DOI: 10.1039/D0CC02437F

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Chemical Communications

The engineering of decameric d-fructose-6-phosphate aldolase A by combinatorial modulation of inter- and intra-subunit interactions†

Abstract

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The engineering of decameric D-fructose-6-phosphate aldolase A by combinatorial modulation of inter- and intra-subunit interactions

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