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Heterologous Expression of Enzymes for Trehalose Synthesis from Sulfolobus acidocaldarius in Bacillus subtilis

WANG Shanying, LI Youran, GU Zhenghua, ZHANG Liang, DING Zhongyang, SHI Guiyang*   

  1. 1. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China;
    2. School of Biotechnology, Jiangnan University, Wuxi 214122, China
  • Online:2016-11-15 Published:2016-11-18

Abstract:

Trehalose has great potential applications in both the medical and food industries due to its unique properties.
Enzymatic transformation is superior to extraction from yeast cells for the industrial production of trehalose. Starch can
be transformed into trehalose via maltooligosylterhalose synthase (MTSase) and maltooligosyltrehalose trehalohydrolase
(MTHase) from Sulfolobus acidocaldarius. In this experiment, we constructed six recombinant expression plasmids
harboring the codon-optimized MTSase and MTHase encoding genes from Sulfolobus acidocaldarius, which could be
expressed under the control of the xylose operons from Bacillus subtilis, Bacillus megaterium and Bacillus licheniformis.
The recombinant plasmids were transformed into Bacillus subtilis, respectively. The effects of different culture conditions
on the production of trehalose was investigated and the optimal culture conditions were determined as follows: glycerol
concentration as the best carbon source, 5 g/L; peptone concentration as the best nitrogen source, 24 g/L; xylose
concentration, 6 g/L; incubation duration, 8 h; and induction temperature, 37 ℃; induction time, 16 h. Experiments carried
out under these conditions gave a conversion rate of trehalose of 33.57%. This study has successfully achieved the functional
expression of MTSase and MTHase in Bacillus subtilis.

Key words: shuttle vector, Sulfolobus acidocaldarius, Bacillus subtilis, inducible expression

CLC Number: