• Title/Summary/Keyword: ABA hydroxylation

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Heterologous Expression of the Hot Pepper ABA 8'-Hydroxylase in Escherichia coli for Phaseic Acid Production

  • Hyun Min Kim;Young Hee Joung
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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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.

Effects of Foliar-sprayed Diniconazole on Contents of Endogenous Gibberellic Acids and Abscisic Acid in Lilium davuricum (Diniconazole 엽면살포가 날개하늘나리의 내생 GA 및 ABA 함량에 미치는 영향)

  • Eum, Sun-Jung;Park, Kyeung-Il;Lee, In-Jung;Choi, Young-Jun;Oh, Wook;Kim, Kiu-Weon
    • Horticultural Science & Technology
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    • v.29 no.3
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    • pp.165-171
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    • 2011
  • Plant growth retardants reduce the plant height by inhibiting stem elongation in Lilium davuricum. To investigate the plant hormones related to stem elongation, we sprayed 50 $mg{\cdot}L^{-1}$ diniconazole to young plants of L. davuricum and quantified the contents of endogenous gibberellic acids (GA) and abscisic acid (ABA). In GA biosynthesis, L. davuricum had not only the early C-13 hydroxylation ($GA_{19}{\rightarrow}GA_{20}{\rightarrow}GA_1$) pathway resulting in $GA_1$ as the active form but also the non C-13 hydroxylation (NCH, $GA_{12}{\rightarrow}GA_{24}{\rightarrow}GA_9{\rightarrow}GA_4$) with $GA_4$ as the active form. However, the main pathway was NCH because $GA_4$ concentration of 55 $ng{\cdot}g^{-1}$ dry wt was much higher than $GA_1$ content of 0.23 $ng{\cdot}g^{-1}$ dry wt in control plant. Diniconazole inhibited GA biosynthesis through NCH pathway from its early stage. $GA_{12}$ content decreased by diniconazole up to 6% level of that of control and this effect continued to $GA_4$. Diniconazole reduced $GA_{12}$ content by 12.7 $ng{\cdot}g^{-1}$ dry wt, whereas that of control plant was 213.8 $ng{\cdot}g^{-1}$ dry wt. ABA content decreased up to one third of control by diniconazole application. From the contents of endogenous $GA_4$, $GA_1$, and ABA in this study, we could conclude that diniconazole reduces the plant height by inhibiting $GA_4$ biosynthesis in L. davuricum.