期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 62, 期 13, 页码 2913-2918出版社
AMER CHEMICAL SOC
DOI: 10.1021/jf405694n
关键词
Tat pathway; twin-arginine signal peptide; methyl parathion hydrolase; Bacillus subtilis 168
资金
- National High Technology Research and Development Program of China (863 Program) [2013AA06A210]
- National Natural Science Foundation of China [31300032]
- Open Project Funding of the State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University [MMLKF13-06]
- Project of Tianjin, China [13JCQNJC09700]
- Open Project Funding of the Key Laboratory of Original Agro-environment Quality of Ministry of Agriculture and Tianjin Key Laboratory of Agro-environment and Safe-product [2013-KF-06]
The twin-arginine translocation (Tat) pathway exports folded proteins across the cytoplasmic membranes of bacteria and archaea. Two parallel Tat pathways (TatAdCd and TatAyCy systems) with distinct substrate specificities have previously been discovered in Bacillus subtilis. In this study, to secrete methyl parathion hydrolase (MPH) into the growth medium, the twin-arginine signal peptide of B. subtilis YwbN was used to target MPH to the Tat pathway of B. subtilis. Western blot analysis and MPH assays demonstrated that active MPH was secreted into the culture supernatant of wild-type cells. No MPH secretion occurred in a total-tat, mutant, indicating that the observed export in wild-type cells was mediated exclusively by the Tat pathway. Export was fully blocked in a tatAyCy mutant. In contrast, the tatAdCd mutant was still capable of secreting MPH. These results indicated that the MPH secretion directed by the YwbN signal peptide was specifically mediated by the TatAyCy system. The N-terminal sequence of secreted MPH was determined as AAPQVR, demonstrating that the YwbN signal peptide had been processed correctly. This is the first report of functional secretion of a heterologous protein via the B. subtilis TatAyCy system. This study highlights the potential of the TatAyCy system to be used for secretion of other heterologous proteins in B. subtilis.
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