公开(公告)号：US20220204965A1

公开(公告)日：2022-06-30

申请号：US17525283

申请日：2021-11-12

申请人： Qilu University of Technology

发明人： Hongling Liu ,  Tengfei Wang ,  Ruiming Wang ,  Xinyi Zhang

IPC分类号： C12N11/18 ,  C12N15/75 ,  C12N9/10 ,  C12N9/26 ,  C12N9/90

摘要： The present disclosure relates to a method for constructing a trehalose polyenzyme complex in vitro by mediation of an artificial scaffold protein, which mainly comprises the following steps: constructing a recombinant bacterium WB800n-ScafCCR for self-assembled scaffold protein module; constructing a recombinant bacterium WB800n-P43-phoD-treY-Ccdoc for self-assembled catalytic module; constructing a recombinant bacterium WB800n-P43-phoD-treZ-Ctdoc for self-assembled catalytic module; constructing a recombinant bacterium WB800n-P43-phoD-cgt-Rfdoc for self-assembled catalytic module; secretorily expressing the recombinant bacteria and self-assembling in vitro to obtain a recombinant trehalose multi-enzyme complex. The trehalose multi-enzyme complex constructed by the method of the present disclosure has a higher catalytic efficiency in preparing trehalose than that of mixed free enzymes, and the method can be used for production of high quality trehalose after immobilization with cellulose microspheres.

公开(公告)号：US12043834B2

公开(公告)日：2024-07-23

申请号：US17525283

申请日：2021-11-12

申请人： Qilu University of Technology

发明人： Hongling Liu ,  Tengfei Wang ,  Ruiming Wang ,  Xinyi Zhang

IPC分类号： C12N15/52 ,  C12N9/10 ,  C12N9/26 ,  C12N9/90 ,  C12N11/18 ,  C12N15/03 ,  C12N15/75

CPC分类号： C12N15/52 ,  C12N9/1051 ,  C12N9/2408 ,  C12N9/90 ,  C12N11/18 ,  C12N15/03 ,  C12N15/75 ,  C12Y302/01141 ,  C12Y504/99015

摘要： The present disclosure relates to a method for constructing a trehalose polyenzyme complex in vitro by mediation of an artificial scaffold protein, which mainly comprises the following steps: constructing a recombinant bacterium WB800n-ScafCCR for self-assembled scaffold protein module; constructing a recombinant bacterium WB800n-P43-phoD -treY-Ccdoc for self-assembled catalytic module; constructing a recombinant bacterium WB800n-P43-phoD-treZ-Ctdoc for self-assembled catalytic module; constructing a recombinant bacterium WB800n-P43-phoD-cgt-Rfdoc for self-assembled catalytic module; secretorily expressing the recombinant bacteria and self-assembling in vitro to obtain a recombinant trehalose multi-enzyme complex. The trehalose multi-enzyme complex constructed by the method of the present disclosure has a higher catalytic efficiency in preparing trehalose than that of mixed free enzymes, and the method can be used for production of high quality trehalose after immobilization with cellulose microspheres.