IPro, PLLC
Construction and application of engineering bacteria capable of secreting and expressing diacetylchitobiose deacetylase
The disclosure herein relates to construction and application of engineering bacteria capable of secreting and expressing diacetylchitobiose deacetylase, and belongs to the technical field of fermentation engineering. Firstly, recombinant B. subtilis capable of heterologously secreting and expressing a diacetylchitobiose deacetylase gene is constructed, and a signal peptide fragment yncM is added into the recombinant vector for the first time. The signal peptide can secrete the target protein diacetylchitobiose deacetylase outside the cells of the recombinant B. subtilis, and a mutant of the 5′-end untranslated region is acquired, thereby significantly increasing the expression level of the target protein, and greatly simplifying the subsequent enzyme separation and purification steps. When the acquired diacetylchitobiose deacetylase is fermented and cultured in a fermentation medium for 50-60 h, the enzyme activity reaches a maximum of 1,548.7 U/mL, and the maximum yield of the diacetylchitobiose deacetylase is about 620 mg/L. Simultaneously, the method has the advantages of low production cost, mild production conditions, simple purification process steps, safe production operation and the like.
SHANDONG RUNDE BIOTECHNOLOGY CO., LTD Jiangnan University
C C C A A C A C C
C12N C12N C12N A23L A61K C12P A23L C12N C12N
additional inventional inventional inventional inventional inventional inventional additional inventional
Wei Jianghua Long Hongzhi Zhu Xueqin Jianxing Guocheng Jian Changfeng
Lu Li Liu Zhang Jiang Lv Lu Du Chen Liu