Modification Strategy of Enzyme Thermal Stability Based on Sequence and Structure Analysis

doi:10.13523/j.cb.2105021

China Biotechnology

2021, Vol. 41

Issue (10): 100-108 DOI: 10.13523/j.cb.2105021

Modification Strategy of Enzyme Thermal Stability Based on Sequence and Structure Analysis

MING Yue¹,ZHAO Zi-tong¹,WANG Hong-lei²,LIANG Zhi-hong^1,^3,^**(

)

1 College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083, China
2 Yantai Institute of China Agricultural University, Yantai 264670, China
3 Key Laboratory of Safety Assessment of Genetically Modified Organism, Ministry of Agriculture and Rural Affairs,Beijing 100083, China

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Abstract

Functional enzymes have been widely used in the fields of food, chemicals, medicine, etc. However, high temperature reduces the catalytic efficiency of enzymes. The protein engineering technology can be used as a key link to modify enzymes’ structure and function, and to obtain industrial enzymes with thermostability. Traditional directed evolution methods can only rely on random mutations for manual screening with low efficiency. Rational design, as the main method of thermostability modification, can be used to predict potential mutation sites with various computer programs and software, but it requires deep understanding of the catalytic and thermal stability mechanism. For most natural enzymes, it is easy to obtain sequence and crystal structure, and is also an important basis for predicting function. This paper focuses on the modification strategies as follows: common mutation, the mutation based on amino acid preference, trunking of flexible regions, the optimization of intramolecular interactions, the modification of catalytic active regions and computer-aided design with the sequence and crystal structure analysis. These strategies have the advantages of high screening efficiency and modification accuracy, and strong practicability. And it also analyzes the thermostability modification cases of different enzymes, aiming to provide an effective reference for the selection of modification strategies, and also give theoretical support for the heat resistance research of industrial enzymes.

Key words： Sequence analysis Structural analysis Thermostability Modification strategies

Received: 12 May 2021 Published: 08 November 2021

ZTFLH:

TQ033

Corresponding Authors: Zhi-hong LIANG E-mail: lzh105@cau.edu.cn

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	Yue MING
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Cite this article:

MING Yue,ZHAO Zi-tong,WANG Hong-lei,LIANG Zhi-hong. Modification Strategy of Enzyme Thermal Stability Based on Sequence and Structure Analysis. China Biotechnology, 2021, 41(10): 100-108.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2105021 OR https://manu60.magtech.com.cn/biotech/Y2021/V41/I10/100

Table 1 Residual substitution bias of thermophilic and thermophilic proteins in 16 families

Table 2 The position bias of different amino acids for β-turns

Table 3 Improving effect of intramolecular interaction on enzyme thermal stability

Fig.1 RMSF values of E.coli transketolase at different temperatures ( 300 K, 340 K, 370 K)


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