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

HAN Chang;SU Ling-qia;WU Jing;State Key Laboratory of Food Science and Technology,Jiangnan University,School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education,Jiangnan University;

maltooligosyltrehalose synthase;;Bacillus subtilis;;recombinant expression;;fermentation optimization

To obtain the recombinant expression of gene tre Y for maltooligosyltrehalose synthase(MTSase)from Sulfolobusacidocaldarius ATCC 33909 in Bacillus subtilis,the target gene was PCR-amplified using plasmid p ET-24a(+)-tre Y as template,andligated with the expression vector pHY300 PLK,then transformed into the expression host Bacillus subtilis CCTCC M 2016536. The activityof MTSasereached 17.5 U/m L after cultivated in TB culture for 48 h. By single factor test(nitrogen source,nitrogen proportion,nitrogenconcentration,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 nitrogensource(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/m L,which was 2.4 times of that before optimization.