METHOD FOR CONSTRUCTING TREHALOSE MULTI-ENZYME COMPLEX IN VITRO MEDIATED BY ARTIFICIAL SCAFFOLD PROTEIN
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United States of America
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Jun 30, 2022
app pub date -
Nov 12, 2021
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Nov 12, 2021
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Abstract
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.
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