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    Recombinant Bacillus subtilis engineered bacterium capable of efficiently expressing keratinase
    4.
    发明授权
    Recombinant Bacillus subtilis engineered bacterium capable of efficiently expressing keratinase 有权

    公开(公告)号：US11384349B2

    公开(公告)日：2022-07-12

    申请号：US17104122

    申请日：2020-11-25

    申请人： Jiangnan University

    发明人： Juan Zhang ,  Zheng Peng ,  Jian Chen ,  Guocheng Du ,  Xinzhe Mao ,  Hengrui Zhou

    IPC分类号： C12N9/54 ,  C12N1/20 ,  C12R1/125

    摘要： The disclosure discloses a recombinant Bacillus subtilis engineered bacterium capable of efficiently expressing keratinase, and belongs to the technical fields of genetic engineering and fermentation engineering. The disclosure successfully constructs a genetically engineered bacterium B. subtilis WB600-pP43NMK-ker capable of efficiently expressing keratinase by using a keratinase gene from Bacillus licheniformis (BBE11-1) as a target gene, pP43NMK as an expression vector and B. subtilis WB600 as an expression host; and meanwhile, the disclosure conducts in-depth research on fermentation media and fermentation conditions when the engineered bacteria are configured as a production strain to produce keratinase to obtain a fermentation medium capable of increasing a yield of keratinase and an optimum process for fermentation production of keratinase.

    Production-line-type High-throughput Screening System
    5.
    发明申请
    Production-line-type High-throughput Screening System 审中-公开

    公开(公告)号：US20190004076A1

    公开(公告)日：2019-01-03

    申请号：US15708706

    申请日：2017-09-19

    申请人： Jiangnan University

    发明人： Jingwen Zhou ,  Jian Chen ,  Jun Fang ,  Meng Ning ,  Guocheng Du ,  Weizhu Zeng ,  Qiuju Zhang ,  Xiaomei Cao

    IPC分类号： G01N35/02 ,  G01N35/04 ,  G01N35/10 ,  G01N35/00

    摘要： The present invention provides a production-line-type high-throughput screening system, which relates to the field of biotechnology and testing equipment. The system comprises of four manipulators, three parallel conveyor belts with fixed slots, 2-DOF slipway and fixed fixtures, 96-channel pipetting system, coloring device, oscillating mixing device, microplate reader, well plates loading platform and well plates recycling platform. Manual operation takes five minutes to detect one 96-well plate, while this system can handle 20 96-well plates per minute. It expands the number of screening targets, making the screening process more clearly and concisely and liberating manual labor. The system makes effective contributions to the development of microbial breeding technology.

    Construction and application of engineering bacteria capable of secreting and expressing diacetylchitobiose deacetylase
    10.
    发明授权
    Construction and application of engineering bacteria capable of secreting and expressing diacetylchitobiose deacetylase 有权

    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.