Recombinant Expression and Fermentation Optimization of Sulfolobus acidocaldarius ATCC 33909 Maltooligosyltrehalose Synthase in Bacillus subtilis

doi:10.13560/J.cnki.biotech.bull.1985.2017-0086

Abstract

Abstract: To obtain the recombinant expression of gene treY for maltooligosyltrehalose synthase（MTSase）from Sulfolobus acidocaldarius ATCC 33909 in Bacillus subtilis，the target gene was PCR-amplified using plasmid pET-24a（+）-treY as template，and ligated with the expression vector pHY300PLK，then transformed into the expression host Bacillus subtilis CCTCC M 2016536. The activity of MTSasereached 17.5 U/mL after cultivated in TB culture for 48 h. By single factor test（nitrogen source，nitrogen proportion，nitrogen concentration，carbon source，glucose concentration，initial pH，and induction temperature）and orthogonal test（nitrogen concentration，glucose concentration，initial pH，and induction temperature），the optimal fermentation condition was determined as：48.0 g/L of nitrogen source（industrial peptone∶cottonseed powder=3∶1），10.0 g/L of glucose，initial pH of medium=7.0，and the optimal temperature was 30℃. Under optimal conditions，the production of MTSase reached 41.5 U/mL，which was 2.4 times of that before optimization.

Key words: maltooligosyltrehalose synthase, Bacillus subtilis, recombinant expression, fermentation optimization optimization

HAN Chang ,SU Ling-qia, WU Jing. Recombinant Expression and Fermentation Optimization of Sulfolobus acidocaldarius ATCC 33909 Maltooligosyltrehalose Synthase in Bacillus subtilis[J]. Biotechnology Bulletin, 2017, 33(7): 162-168.

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