公开(公告)号：US20230265411A1

公开(公告)日：2023-08-24

申请号：US18187984

申请日：2023-03-22

Applicant: Jiangnan University

Inventor： Tao ZHANG ,  Mengying Hu ,  Bo Jiang ,  Mengli Li

IPC: C12N9/90 ,  C12N15/75

CPC classification number: C12N9/90 ,  C12N15/75

Abstract: The present disclosure discloses a strain producing D-allulose 3-epimerase and application thereof, and belongs to the technical field of bioengineering. The present disclosure provides a method for improving the expression of D-allulose 3-epimerase by screening promoters and optimizing RBS thereof. The recombinant Bacillus subtilis constructed using thevectors pP43NMK-hag and pP43NMK-hag-RBS4 provided by the present disclosure improves the enzyme activity of a target gene D-allulose 3-epimerase, and theenzyme activities in shake flasks upon transformation are 1.30 times and 1.69 times that of an original vector. The present disclosure further provides a non-antibiotic resistance vector and a non-antibiotic resistance recombinant B. subtilis strain. Using the non-antibiotic resistance strain B. subtilis 1A751-dal-/pP43NMK-hag-RBS4-dpe-dal provided by the present disclosure, the highest fermentation enzyme activity in a shake flask is 24.72 U/mL, and the enzyme activity in a fermenter is 714.8 U/mL.

公开(公告)号：US11970723B2

公开(公告)日：2024-04-30

申请号：US18187984

申请日：2023-03-22

Applicant: Jiangnan University

Inventor： Tao Zhang ,  Mengying Hu ,  Bo Jiang ,  Mengli Li

IPC: C12N9/90 ,  C12N15/75

CPC classification number: C12N9/90 ,  C12N15/75

Abstract: The present disclosure discloses a strain producing D-allulose 3-epimerase and application thereof, and belongs to the technical field of bioengineering. The present disclosure provides a method for improving the expression of D-allulose 3-epimerase by screening promoters and optimizing RBS thereof. The recombinant Bacillus subtilis constructed using thevectors pP43NMK-hag and pP43NMK-hag-RBS4 provided by the present disclosure improves the enzyme activity of a target gene D-allulose 3-epimerase, and theenzyme activities in shake flasks upon transformation are 1.30 times and 1.69 times that of an original vector. The present disclosure further provides a non-antibiotic resistance vector and a non-antibiotic resistance recombinant B. subtilis strain. Using the non-antibiotic resistance strain B. subtilis 1A751-dal-/pP43NMK-hag-RBS4-dpe-dal provided by the present disclosure, the highest fermentation enzyme activity in a shake flask is 24.72 U/mL, and the enzyme activity in a fermenter is 714.8 U/mL.