Cloning, expression and properties of trehalase from <i>Pectobacterium cypripedii</i>.

Han Gao; Jinsong Gong; Zikai Wang; Chang SU; Zhenghong XU; Jinsong Shi

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Cloning, expression and properties of trehalase from Pectobacterium cypripedii.

Author: Han GAO ¹ ; Jinsong GONG ¹ ; Zikai WANG ¹ ; Chang SU ¹ ; Zhenghong XU ² ; Jinsong SHI ³
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1. Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
2. National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
3. School of Life Sciences and Health Engineering, Jiangnan University, Wuxi 214122, Jiangsu, China.
Publication Type:Journal Article
Keywords: Pectobacterium cypripedii; ethanol tolerance; trehalase; trehalose
MeSH: Trehalase/metabolism*; Trehalose/metabolism*; Escherichia coli/metabolism*; Ethanol/metabolism*; Cloning, Molecular
From: Chinese Journal of Biotechnology 2022;38(12):4658-4668
CountryChina
Language:Chinese
Abstract: Trehalase is widely used in industrial fermentation, food, medicine and other fields. There is a lack of industrial varieties of trehalase with excellent performance in China. Moreover, the applied research on trehalase was not well conducted. In this study, a strain of Pectobacterium cypripedii was screened from nature, and the gene PCTre encoding an acidic trehalase was cloned and expressed in E. coli BL21(DE3). The highest enzyme activity reached 4130 U/mL after fermenting in a 5 L fermenter for 28 h. The enzymatic properties study showed that PCTre hydrolyzed trehalose specifically. The optimum pH and temperature were 5.5 and 35 ℃, respectively. 80% of the enzyme activity was retained after being treated at pH 4.0, 4.5, and 5.0 for 8 h, showing good acid tolerance. Moreover, it has good tolerance to organic solvents, 60% enzyme activity was retained after being treated with 20% (V/V) ethanol solution for 24 h. Furthermore, trehalose could be completely hydrolyzed within 16 h in a simulated fermentation system containing 20% (V/V) ethanol and 7.5% trehalose, with 500 U/L PCTre added. This indicated a good application potential for industrial ethanol fermentation.