P450 - RECOMBINANT VECTOR AND METHOD FOR PRODUCING RECONSTITUTED CYTOCHROME P450 OXYGENASE-REDUCTASE FUSION PROTEIN USING THE SAME

• Main Authors: KIM GEUN JOONG, CHEONG DAE EUN, LE THIEN KIM, CHOI HYE JI, YUN CHUL HO, YOO SU KYOUNG
• Format: Patent
• Language: English; Korean
• Published: 13.05.2021
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The present invention relates to a recombinant vector and a method for preparing a reconstituted cytochrome P450 oxygenase-reductase fusion protein using the same, wherein after independently expressing oxygenase and reductase in a host cell into which the recombinant vector is introduced, by reassembling them by protein splicing of the split intein, the content of heme contained in the active site of oxygenase can be increased, and enzyme activity and stability can be improved. 본 발명은 재조합 벡터 및 이를 이용한 재조립 시토크롬 P450 옥시게나아제-리덕타아제 융합 단백질의 제조 방법에 관한 것으로, 재조합 벡터가 도입된 숙주세포에서 옥시게나아제와 리덕타아제를 독립적으로 발현시킨 후, 이들을 스플릿 인테인의 단백질 스플라이싱에 의해 재조립함으로써, 옥시게나아제의 활성 부위에 포함되는 헴 함량을 증가시키고, 효소 활성 및 안정성을 향상시킬 수 있다.