Biotechnology Bulletin ›› 2017, Vol. 33 ›› Issue (4): 177-184.doi: 10.13560/j.cnki.biotech.bull.1985.2017.04.023

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Recombinant Expression and Characterization of Trehalase Tre F from Escherichia coli str. K-12 substr. MG1655

YU Lin-gang1, SU Ling-qia2, WU Jing1, 2   

  1. 1. State Key Laboratory of Food Science and Technology,Jiangnan University,Wuxi 214122;
    2. School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education,Jiangnan University,Wuxi 214122
  • Received:2016-09-30 Online:2017-04-25 Published:2017-04-25

Abstract: Trehalase is a trehalose hydrolase,which can distinctively catalyze trehalose into two molecules of glucose. In order to have a recombinant expression and application of the trehalase gene Tre F from Escherichia coli str. K-12 substr. MG1655 in E. coliBL21(DE3),the Tre F was amplified by PCR,and a recombinant strain E. coli BL21(DE3)/pET-24a(+)-Tre F was constructed. The highest activity reached 107 U/mL when the strain was cultured in shake flask for 24 h at induction temperature 25℃and IPTG induction concentration 0.4 mmol/L. Further study of the enzymatic properties indicated that the optimal pH and temperature was 7.0 and 50℃,respectively. The crude enzyme was used in the hydrolysis of trehalose. The reaction conditions for the conversion was as the following:initial trehalose concentration 300 g/L,initial pH 7.0,reaction temperature 30℃,enzyme concentration 84 U/g. When the reaction was performed under this specified condition,the glucose yield reached the maximum of 98.4 % at 36 h. This paper is the first report on the recombinant expression of trehalase gene Tre F of E. coli str. K-12 substr. MG1655 in E. coli BL21(DE3).

Key words: trehalase, gene cloning, recombinant expression, enzyme properties