US 12,084,648 B2
Recombinant bacterium with a high PHA yield and the construction method thereof
Jin Yin, Shenzhen (CN); Yu Wang, Shenzhen (CN); Huayu Zhang, Shenzhen (CN); and Yingzi Li, Shenzhen (CN)
Assigned to Shenzhen Bluepha Biosciences Co., Ltd., Shenzhen (CN)
Filed by Shenzhen Bluepha Biosciences Co., Ltd., Shenzhen (CN)
Filed on Mar. 9, 2023, as Appl. No. 18/181,301.
Claims priority of application No. 202210280731.0 (CN), filed on Mar. 22, 2022.
Prior Publication US 2023/0303966 A1, Sep. 28, 2023
Int. Cl. C12N 1/20 (2006.01); C12N 15/78 (2006.01); C12P 7/625 (2022.01)
CPC C12N 1/205 (2021.05) [C12N 15/78 (2013.01); C12P 7/625 (2013.01)] 4 Claims
 
1. A recombinant Ralstonia eutropha,
wherein a promoter for upregulating a phaJ gene is introduced into a genome of the recombinant Ralstonia eutropha,
wherein a sequence of the promoter for upregulating the phaJ gene is a phaJ43 promoter sequence of SEQ ID NO: 3,
wherein original genes phaC and proC in the genome of the recombinant Ralstonia eutropha are inactivated, a sequence of the original phaC gene is represented by SEQ ID NO: 1, a sequence of the original proC gene is represented by SEQ ID NO: 2,
wherein a plasmid capable of stably existing in Ralstonia eutropha is introduced into the recombinant Ralstonia eutropha, wherein the plasmid is loaded with an introduced phaC mutant gene and an introduced proC gene, wherein a sequence of the introduced phaC mutant gene is represented by SEQ ID NO: 5, the expression of the introduced phaC mutant gene enables the recombinant Ralstonia eutropha to synthesize 3-hydroxybutyrate-co-3-hydroxyhexanoate, a sequence of the introduced proC gene is represented by SEQ ID NO: 4,
wherein an original strain of the recombinant Ralstonia eutropha is Ralstonia eutropha Re0980, and a deposit number for the Ralstonia eutropha Re0980 is GDMCC No: 62177.