• 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.

• Journal of Bio-Environment Control
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• v.31 no.3
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• pp.170-179
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• 2022

Glutamic acid is a precursor of essential amino acids that play an important role in plant growth and development. It is one of the biostimulants that reduce cold stress damage by stimulating biosynthetic pathways leading to cryoprotectants. This study evaluated the effects of glutamic acid foliar application on Kimchi cabbage under low-temperature stress. There were six treatments, combining three photo-/dark periods temperature levels (11/-1℃ extremely low, E; 16/4℃ moderately low, M; and 21/9℃ optimal, O) with and without glutamic acid foliar application (0 and 10 mg·L^-1; Glu 0 and Glu 10). Glutamic acid foliar application was sprayed once 10 days after transplanting, and then temperature treatment immediately after glutamic acid foliar application was conducted for up to four days. After four days of treatment, abscisic acid (ABA), phaseic acid (PA), dihydrophaseic acid (DPA), and abscisic acid-glucose ester (ABA-GE) contents were higher with Glu 10 treatment than Glu 0 treatment in M treatment. Glucose content was highest in E with Glu 10 treatment (52.1 mg·100 g^-1 dry weight), while fructose content was highest in O with Glu 0 treatment (134.6 mg·100 g^-1 dry weight). The contents of glucolepiddin (GLP), glucobrassicin (GBS), 4-methoxyglucobrassicin (4MGBS), neoglucobrassicin (GNBS), and gluconasturtiin (GNS) were highest among all treatments in E with Glu 10 treatments (0.72, 2.05, 1.67, 9.40 and 0.85 µmol·g^-1 dry weight). After two days of treatment, rapid changes in PA and DPA contents of E with Glu 10 treatments were confirmed, and several individual glucosinolate contents (GLP, GBS, 4MGBS, GNBS, and GNS) were significantly different depending on low temperature and glutamic acid treatment. In addition, the content of fructose was significantly lower than that of O treatment in E and M treatments after four days of treatment. Therefore, although the changes in PA, DPA, glucose, fructose, and individual glucosinolates according to low temperature and glutamic acid foliar treatment were shown. A clear correlation between low temperature and glutamic acid effects could not be evaluated. Results indicated that Brassica crops are cryophilic vegetables, do not react sensitively to low temperatures, and mostly have cold resistance.