More efficient enzymatic cascade reactions by spatially confining enzymes via the SpyTag/SpyCatcher technology

Journal article (2022)

Authors

Xuanru Zhong South China University of Technology
Yunjian Ma South China University of Technology, Macau University of Science and Technology
Xizhen Zhang South China University of Technology
Jiahui Zhang South China University of Technology
Bin Wu South China University of Technology
F. Hollmann BT/Biocatalysis - AS
Yonghua Wang Guangdong Youmei Institute of Inteligent Bio-manufacturing Co, Guangdong, South China University of Technology
Research Group
BT/Biocatalysis (AS) (TU Delft)
Copyright: © 2022 Xuanru Zhong, Yunjian Ma, Xizhen Zhang, Jiahui Zhang, Bin Wu, F. Hollmann, Yonghua Wang
DOI
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https://doi.org/10.1016/j.mcat.2022.112188
CvFAP Hydrocarbon biofuels Multienzyme complex SpyTag/SpyCatcher TLL
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Published Date

2022

Language

English

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Faculty

Applied Sciences

Department

BT/Biotechnologie

Research Group

BT/Biocatalysis

Abstract

Hydrocarbon synthesis from (waste)oils enabled by a cascade of lipase-catalysed hydrolysis and decarboxylase-catalysed decarboxylation has become an active area of research en route to alternative, biobased fuels. However, Poor substrate transport efficiency is a major issue causing low reaction rates. This study focused on a protein self-assembly strategy based on SpyTag/SpyCatcher to overcome diffusion limitations. For this, two fusion proteins, TLL-Linker-SpyCatcher based on the lipase from Thermomyces lanuginosus and CvFAP-Linker-SpyTag based on the fatty acid photodecarboxylase from Chlorella variabilis were designed. A covalent multi-enzyme complex (TLL-CvFAP) was formed spontaneously by self-assembly of each enzyme. The effects of temperature, pH and molar ratio of self-assembled components on assembly efficiency were investigated. The results showed that the multi-enzyme complex TLL-CvFAP reached about 60% after 12 h of assembly, and the enzyme activity of the multienzyme complex was increased by about 50% compared to that of the corresponding non-assembled enzymes. Under optimized conditions 10 mM soybean oil were converted into 25 mM of the corresponding hydrocarbons, suggesting a good potential of biofuel synthesis.