Expression and secretion of a single-chain sweet protein monellin in <Emphasis Type="Italic">Bacillus subtilis</Emphasis> by <Emphasis Type="Italic">sacB</Emphasis> promoter and signal peptide |
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| Authors: | Zhongjun Chen Cai Heng Zhengying Li Xinle Liang Shangguan Xinchen |
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| Institution: | (1) College of Food Science and Engineering, Inner Mongolia Agricultural University, 010018 Huhhot, People’s Republic of China;(2) College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, 210009 Nanjing, People’s Republic of China;(3) Department of Biology Engineering, Zhejiang Gongshang University, 310035 Hangzhou, People’s Republic of China;(4) College of Food Science and Engineering, Jiangxi Agricultural University, 330045 Nanchang, People’s Republic of China |
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| Abstract: | The sweet protein monellin gene was expressed in Bacillus subtilis under the control of the Bacillus subtilis
sacB promoter and signal peptide sequence. A 294-bp DNA fragment, coding for sweet protein monellin, was ligated into the Escherichia coli/B. subtilis shuttle vector pHPC, producing pHPMS, which was subsequently transformed into B. subtilis QB1098, DB104, and DB403. The peptide efficiently directed the secretion of monellin from the recombinant B. subtilis cells. A maximum yield of monellin of 0.29 g protein l−1 was obtained from the supernatant of B. subtilis DB403 harboring pHPMS. SDS-PAGE confirmed the purity of the recombinant product. |
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| Keywords: | Bacillus subtilis Expression Monellin Proteases deficiency |
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