公开(公告)号：US10934552B2

公开(公告)日：2021-03-02

申请号：US16503699

申请日：2019-07-05

申请人： Jiangnan University ,  SHANDONG RUNDE BIOTECHNOLOGY CO., LTD

发明人： Long Liu ,  Jian Chen ,  Guocheng Du ,  Xueqin Lv ,  Jianghua Li ,  Zhu Jiang ,  Wei Lu ,  Hongzhi Zhang ,  Jianxing Lu ,  Changfeng Liu

IPC分类号： C12N15/75 ,  C12N1/20 ,  C12N9/80 ,  A61K31/7008 ,  A23L29/30

摘要： 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.