Optimization of Medium Composition for Expression of Recombinant Sucrose Phosphorylase and Effect of Chemical Reagents on Enzyme Activity

ZHANG Hui; ZHOU Xiaomei; ZHANG Biao; SUN Xiao; LI Wenjie; LI Tuoping

doi:10.13386/j.issn1002-0306.2020110137

Volume 42 Issue 15

Jul. 2021

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Science and Technology of Food Industry > 2021 > 42(15): 119-124. > DOI: 10.13386/j.issn1002-0306.2020110137

ZHANG Hui, ZHOU Xiaomei, ZHANG Biao, et al. Optimization of Medium Composition for Expression of Recombinant Sucrose Phosphorylase and Effect of Chemical Reagents on Enzyme Activity[J]. Science and Technology of Food Industry, 2021, 42(15): 119−124. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2020110137.

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Optimization of Medium Composition for Expression of Recombinant Sucrose Phosphorylase and Effect of Chemical Reagents on Enzyme Activity

ZHANG Hui^{1, 2,},
ZHOU Xiaomei³,
ZHANG Biao¹,
SUN Xiao⁴,
LI Wenjie^4, ,,
LI Tuoping^2, ,

1.
Boda College of Jilin Normal University, College of Food Science and Engineering, Siping 136000, China
2.
Shenyang Agricultural University, Food College, Shenyang 110866, China
3.
Jilin Normal University, College of Life Sciences, Siping 136000, China
4.
Shenyang Women’s and Children’s Hospital, Shenyang 110011, China

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Received Date: November 16, 2020
Available Online: June 03, 2021

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Abstract

Abstract

In order to improve the expression level of recombinant sucrose phosphorylase, the effects of different carbon sources, nitrogen sources, protectants and metal ions on the expression of recombinant sucrose phosphorylase in Escherichia coli was studied by single factor test, then the optimal composition of recombinant sucrose phosphorylase expression medium was determined by orthogonal test. In addition, the effects of different chemical reagents and metal ions on the stability of recombinant sucrose phosphorylase were investigated.The results showed that vitamin C and FeCl₂ significantly (P<0.01) inhibited the protein expression, and Cu²⁺ and Zn²⁺ completely killed the microorganisms. On the contrary, starch, K⁺ and Mg²⁺ could promote the expression of recombinant sucrose phosphorylase, and the synergistic effect could greatly improve the expression efficiency of recombinant sucrose phosphorylase. The total enzyme activity of optimal combination, with 0.5% starch, 0.1% K⁺ and 0.05% Mg²⁺ in 50 mL LB medium, was 1207.83 U, which was increased by 3.6 -fold. The results showed that 10 mmol/L Fe²⁺, 10 mmol/L Mg²⁺, 1% bovine serum protein and 0.1% Tween 80 could increase the enzyme activity and stability of sucrose phosphorylase in a certain extent.
- sucrose phosphorylase,
- medium optimization,
- carbon and nitrogen sources,
- chemical reagent,
- metal ion

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