• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1111-1119
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• 2006

Beauvericins and enniatins are cyclohexadepsipeptides exhibiting various biological activities on animal systems, including humans. Fusarium oxysporum FB1501 (KFCC 11363P) that produces four different cyclohexadepsipeptides was isolated from soil in Korea and the structures of the four cyclohexadepsipeptides elucidated by HPLC, MS, IR, and NMR analyses. The molecular weights for compounds 1,2,3, and 4 were determined to be 654.5, 784.5, 668.6, and 682.5, respectively, on the basis of ESI-MS measurements. The IR spectra for all the compounds exhibited absorptions for ester $(1,733-1,743\;cm^{-1})$ and amide $(1,649-1,655\;cm^{-1})$ bonds that were very similar to those for beauvericin and enniatins with ester and amide absorptions. The results of the NMR analysis $(^{1}H,\;^{13}C,\;135-DEPT,\;COSY,\;HMQC,\;and\;HMBC;\;in\;COCl_{3})$ revealed that compounds 1,3, and 4 consisted of $_{L}-N-methyl\;valine$ (N-MeVal), $_{D}-{\alpha}-hydroxyisovaleic\;acid$ (Hiv), and 2-hydroxy-3-methylpentanoic acid (Hmp) residues (compound 1: three N-MeVal residues, two Hiv residues, and one Hmp residue; compound 3: three N-MeVal residues, one Hiv, and two Hmp residues; compound 4: three N-MeVal residues and three Hmp residues). Therefore, the compounds were identified as enniatin H (compound 1), enniatin I (compound 3), and enniatin MK1688 (compound 4). Compound 2 was analyzed as beauvericin according to 1D and 2D NMR analyses. This study is the first report related to the co-production of beauvericin with other unusual enniatins, such as enniatin H, enniatin I, and enniatin MK1688, by Fusarium oxysporum.

• Journal of Food Hygiene and Safety
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• v.37 no.5
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• pp.347-355
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• 2022

Levels of beauvericin (BEA) and enniatins (ENNs: ENA, ENA1, ENB, and ENB1) were examined in fresh ginger (n = 43) and ginger powder (n = 31) samples from Korea. In the ginger samples, incidence of BEA contamination was highest, at 62.79%, with a maximum detected BEA level of 640.07 ㎍/kg. ENNs in were found in up to 11.63% (ENB, ENB1) of ginger samples, with a maximum detected level of 91.02 ㎍/kg (ENA). In the ginger powders, ENB contamination displayed the highest rate of incidence (70.97%), but the highest level of BEA (1,344.18 ㎍/kg) exceeded that of ENB (413.99 ㎍/kg). The incidences of ENA, ENA1, ENB, and ENB1 presence in ginger powders were 29.03%, 22.58%, 70.97%, and 35.48%, respectively, and their highest detected levels were 220.45, 156.61, 413.99, and 70.29 ㎍/kg, respectively. The incidence of BEA and ENN contamination was higher in ginger powder than in ginger. Respective co-occurrence rates of BEA and ENNs in ginger and ginger powder samples were 16.28% and 64.52%, indicating that the BEA and ENN co-contamination rate was highest in ginger powder as well. This is the first report on the presence and co-occurrence of BEA and ENNs in Korean ginger and ginger powder.

• Journal of Food Hygiene and Safety
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• v.23 no.1
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• pp.73-79
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• 2008

The productions of beauvericin and enniatins H, I, and MK1688 by Fusarium oxysporum KFCC 11363P were investigated on rice substrate at four temperatures (15, 20, 25, and $30^{\circ}C$) and three moisture contents (10, 20, and 40%). The largest amount of beauvericin ($718.0\;{\mu}g/g$) was produced at $25^{\circ}C$, and maximum levels of enniatin H ($781.9\;{\mu}g/g$), I ($725.8\;{\mu}g/g$), and MK1688 ($425.8\;{\mu}g/g$) were measured by high pressure liquid chromatography (HPLC) at the same temperature. The optimal moisture content for the production of beauvericin and enniatins H, I, and MK1688 was 40%, and the trace amounts of these toxins were observed at 10% moisture content. Sixty five grain samples (n=65) were tested for the monitoring of beauvericin. This mycotoxin was detected in six grain samples including three maize, two barley, and one wheat samples. The highest contamination level of beauvericin was observed in maize sample ($0.23\;{\mu}g/g$).

• BMB Reports
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• v.33 no.3
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• pp.228-233
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• 2000

The steady-state investigation of the mechanism of Dhydroxyisovalerate dehydrogenase was performed in order to understand this type of kinetic patterns. The initial velocity was measured with various amounts of both substrates, NADPH and 2-ketoisovalerate. Double reciprocal plots gave patterns that conversed on or near the abscissa. Binding studies indicated that NADPH bound first to the enzyme. The product $NADP^+$ was found to be a competitive inhibitor with respect to NADPH at a constant concentration of 2-ketoisovalerate. However, it showed noncompetitive inhibition against 2-ketoisovalerate at a fixed amount of NADPH. Another product, D-hydroxyisovalerate, was a non-competitive inhibitor versus NADPH and 2-ketoisovalerate at constant levels of 2-ketoisovalerate and NADPH, respectively. These results were comparable with an ordered bi-bi mechanism, in which NADPH bound first to the enzyme, followed by the binding of 2- ketoisovalerate. $NADP^+$ is the last product to be released. The ordered reaction manner of D-hydroxyisovalerate dehydrogenase from 2-ketoisovalerate to D-hydroxyisovalerate allows the accurate regulation of valine metabolism and it may lead to the regulation of total biosynthesis of enniatins in the Fusarium species.

• BMB Reports
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• v.29 no.6
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• pp.493-499
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• 1996

The biochemical properties of purified D-hydruxyisovalerate dehydrogenase from Fusarium sambucinum was elucidated. D-Hydroxyisovalerate dehydrogenase produced solely D-hydroxyisovalerate from 2-ketoisovalerate. The isoelectric point of the purified enzyme was 7.0. The enzyme was highly specific with 2-ketoisovalerate ($K_{m}=0.188$ mM, $V_{max}=8.814$ mmol/min mg) and 2-keto-3-methyl-n-valerate ($K_{m}=0.4$ mM, $V_{max}=1.851$ mmol/min mg) for the reductive reaction. This was also seen by comparing D-hydroxyisovalerate ($K_{m}=1.667$ mM, $V_{max}=0.407$ mmol/min mg) and D-hydroxy-3-methyl-n-valerate ($K_{m}=6.7$ mM, $V_{max}=0.648$ mmol/min mg) for the oxidative reaction. Thiol blocking reagents, such as iodoacetamide, N-ethylmaleimide and p-chloromecuribenzoate inhibited about 80% of enzyme activity at 0.02 mM, 50 mM and 50 mM, respectively. The enzyme activity was also inhibited by the addition of 0.1 mM of various metal ions, such as $Fe^{2+}$ (67%), $Cu^{2+}$ (88%), $Zn^{2+}$ t (76%) and $Mg^{2+}$ (9%). The enzyme was stable over three months in 50 mM potassium phosphate buffer (pH 5~7) at $-80^{\circ}C$. However the purified enzyme lost 30% of its activity in the same buffer after 24 h at $4^{\circ}C$. The studies about thermal inactivation of D-hydroxyisovalerate dehydrogenase exhibit 209.2 kJ/M of activation enthalpy and 0.35 kJ/mol K of activation entropy.