Journal of Pure and Applied MicrobiologyVol. 8 No. 2

Cloning and expression of a Trehalose Synthase from Pseudomonas putida KT2440 in Bacillus subtilis W800N for the production of Trehalose

Tengfei Wang1,2*, Kun Dai2, Hongjuan Liu2, Shiru Jia1 and Ruiming Wang2

1Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, TEDA, Tianjin 300457, PR China. 2Key Laboratory of Shandong Microbial Engineering, QILU University of Technology, Jinan, Shandong 250353, PR China.

Received on 28 February 2014 and accepted on 08 April 2014

 

ABSTRACT

Trehalose synthase which catalyze inexpensive maltose to trehalose by one-step hydrolysis reaction, was considered to be a potential biocatalyst for trehalose production. In our work, the DNA encoding trehalose synthase with 6?his-tag was ligated with pGEM-T easy vector, transformed into E.coli DH5? and screened. After sequence analysis, the PCR products was digested by BamHI and AatII and ligated into pHT01 by using T4 DNA ligase to generate the recombinant vector pHT01-TreS. The purified pHT01-TreS was electrotransformed into B.subtilis W800N. The recombinant TreS protein was expressed and purified by nickel affinity chromatogaraphy. The optimum temperature and pH were 25?C and pH 8.0, respectively. The pure enzyme was stable in pH 6.0 to 9.0. It activity increased by Na+, K+ and Mg2+, but it was greater inhibited by Zn2+ and Ca2+.

Keywords : Trehalose synthase, Gene cloning, Protein expression, Bacillus subtilis W800N, Pseudomonas putida KT2440, Conversion.