• Journal of Microbiology and Biotechnology
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• 제33권3호
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• pp.378-386
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• 2023

The CYP707A family genes encoding ABA 8'-hydroxylase catabolize abscisic acid (ABA), a plant stress hormone that plays an important role in stress condition, such as drought, heat, cold and salinity. Phaseic acid (PA) is a catabolic product of ABA. Recent studies have shown that PA is important for the physiological functions in plants. It is also a neuroprotective molecule that protects against ischemic brain injury in mice. To obtain enzymes for the PA production, four CaCYP707A genes (CaCYP707A1, CaCYP707A2, CaCYP707A3 and CaCYP707A4) were isolated from hot pepper. They were heterologously expressed in Escherichia coli. Among them, CaCYP707A2 showed significantly higher expression levels in both the membrane fraction and the soluble fraction. Preferred redox partners were investigated to improve the efficiency of CaCYP707A2's catalytic reaction, and NADPH-cytochrome P450 reductase (CPR) from hot pepper (CaCPR) was preferred over other redox partners (i.e., rat CPR and ferredoxin reductase/ferredoxin). The production of 8'-hydroxy ABA and PA by ABA hydroxylation activity was confirmed in CaCYP707A2 from both membrane and soluble fractions. Therefore, CaCYP707A2 is the first identified plant CYP protein that is expressed a soluble form in cytosolic fraction having stable activity. Taken together, we propose a new CYP707A protein with industrial applications for PA production without additional modifications in E. coli heterologous expression.

• 원예과학기술지
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• 제29권3호
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• pp.165-171
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• 2011

식물생장억제물질의 처리는 날개하늘나리의 줄기 신장을 억제시키는데, 여기에 관여하는 식물호르몬을 찾기 위해 diniconazole의 엽면살포 후 내생 GA 및 ABA의 함량 변화를 조사하였다. 그 결과, 날개하늘나리(L. dauricum)는 $GA_1$을 활성형으로 하는 early C-13 hydroxylation($GA_{19}{\rightarrow}GA_{20}{\rightarrow}GA_1$) 경로와 $GA_4$를 활성형으로 하는 non C-13 hydroxylation(NCH, $GA_{12}{\rightarrow}GA_{24}{\rightarrow}GA_9{\rightarrow}GA_4$) 경로 모두를 가지고 있었으나, 주된 경로는 NCH 경로였다. NCH 경로의 GA 생합성은 diniconazole $50mg{\cdot}L^{-1}$ 살포에 의해 초기 단계에서부터 억제되었다. 즉 diniconazole 처리구의 $GA_{12}$ 함량은 대조구에 비해 1/17로 현저히 감소되었으며, 이러한 경향은 $GA_4$까지 계속되었다. 즉 건물 1g당 $GA_{12}$ 함량은 대조구가 213.8ng인 것에 비해, diniconazole 처리구는 12.7ng이었다. ABA의 함량도 GA에서와 같이 diniconazole 살포에 의해 1/3 수준으로 크게 감소되었다. 즉 건물 1g당 ABA 함량은 대조구 37.2ng인 것에 비해, diniconazole 처리구는 14.8ng이었다. 본 연구의 결과, 내생 $GA_4$, $GA_1$, 그리고 ABA의 함량을 고려할 때 diniconazole에 의한 날개하늘나리의 초장감소는 $GA_4$의 생합성 억제에 기인한 것으로 생각된다.