• Applied Biological Chemistry
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• v.46 no.4
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• pp.361-366
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• 2003

Anti-aflatoxigenic studies on synthetic pyridione alkaloids were conducted. Seven derivatives using piperlongumine as a leading compound were prepared from 3,4,5-trimethoxycinnamic acid (TMCA). These derivatives were analyzed for their structural confrmation and purity by HPLC, GC, GC/MS and $1^H-NMR$. 1-piperidin-1-yl-3-(3,4,5-trimethoxyphenyl)propenone (1) reaction with piperidine; 1-morpholin-4-yl-3-(3,4,5-trimethoypenyl)propenone (2) with morpholine; 1-(3,5-dimethylpiperidin-1-yl)-3-(3,4,5-trimethoxyphenyl)propenone (3) with 3,5-dimethylpiperdine; 1-(2-methylpiperidine-1-yl)-3-(3,4,5-trimethoxyphenyl)propenone (4) with 2-methylpiperidine; 1-(3-hydroxypiperidin-1-yl)-3- (3,4,5-trimethoxyphenyl)propenone (5) with 3-hydroxypiperidine hydrochloride; 1-[3- (3,4,5-trimethoxyphenyl)acryloyl]piperidin-2-one (6) with ${\delta}-valerolactam;\; and\;ethyl\;1-[3-(3,4,5-trimethoxyphenyl)acyloyl]piperidine-4-carboxylate$ (7) with ethyl isonipectotate were synthesized respectively. All derivatives showed an inhibitory activity on aflatoxin $B_1$ production. In conclusion, we believe that they might be an agent for the control of mycotoxin in agricultural commodities.

• Toxicological Research
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• v.15 no.1
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• pp.95-101
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• 1999

Cytochrome P450 (CYP) 3A4 metabolizes aflatoxin B1 (AFB1) to AFB1-exo-8,9-epoxide (8,9-epoxidation) and aflatoxin Q1 (AFQ1; 3$\alpha$-hydroxylation) simultaneously. We investigated whether each metabolite was formed via its own binding site of CAP3A4 active site. Kinetics of the formation of the two metabolites were sigmoidal and consistent with the kinetics of substrate activation. The HIll model predicted that two substrate binding wites are involved in the oxidationof AFB1 by CYP3A4. Dehydronifedipine, a metabolite of nifedipine generated by CYP3A4, inhibited the formation of AFQ1 without any inhibition in the formation of AFB1-exo-8,9-epoxidation. Dehydronifedipine was found to act as a reversible competitive inhibitor against 3$\alpha$-hydroxylation of AFB1. Vmax and S0.5 of the 8,9-epoxidation were not changed in the presence of 0, 50, or 100 $\mu\textrm{M}$ dehydronifedipine. S0.5 of 3$\alpha$-hydroxylation was increased from 58$\pm$4 $\mu\textrm{M}$ to 111$\pm$8 $\mu\textrm{M}$ in the presence of 100 $\mu\textrm{M}$ nifedipine whereas Vmax was not changed. These results suggest that there exist two independent binding sites in the active site of CAP3A4 . One binding site is responsible for AFB1-exo-8,9-epoxidation and the other is involved in 3$\alpha$-hydroxylation of AFB1. Dehydronifedipine might selectively bind to the site which is responsible for the formation of AFQ1 in the active site of CYP3A4.

• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1207-1213
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• 2020

Aflatoxin B1 (AFB1) is a mycotoxin produced by Aspergillus flavus (A. flavus). AFB1 is reported to have high thermal stability and is not decomposed by heat treatment during food processing. Therefore, in this study, knowing that AFB1 is metabolized by cytochrome P450 (CYP), our aim was to develop a method to detoxify A. flavus-contaminated maize, under normal temperature and pressure, using Escherichia coli expressing human CYP3A4. First, the metabolic activity of AFB1 by recombinant human CYP3A4 was evaluated. As a result, we confirmed that recombinant human CYP3A4 metabolizes 98% of AFB1. Next, we found that aflatoxin Q1, a metabolite of AFB1 was no longer mutagenic. Furthermore, we revealed that about 50% of the AFB1 metabolic activity can be maintained for 3 months when E. coli expressing human CYP3A4 is freeze-dried in the presence of trehalose. Finally, we found that 80% of AFB1 in A. flavus-contaminated maize was metabolized by E. coli expressing human CYP3A4 in the presence of surfactant triton X-405 at a final concentration of 10% (v/v). From these results, we conclude that AFB1 in A. flavus-contaminated maize can be detoxified under normal temperature and pressure by using E. coli expressing human CYP3A4.

• 한국균학회소식:학술대회논문집
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• 2006.10a
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• pp.32-44
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• 2006

To investigate non-aflatoxin-production of A. oryzae at the molecular level, an aflatoxin biosynthesis gene homolog cluster of RIB 40 was analyzed. Although most genes in the corresponding cluster exhibited from 97 to 99 % similarity to those of Aspergillus flavus, three genes shared 93 % similarity or less. In addition, although slight expression of aflR, positive transcriptional regulator gene, was detected in some A. oryzae strains having seven aflatoxin biosynthesis homologous genes, other genes related to aflatoxin production were not detected. RIB strains were mainly divided into group 1, having seven aflatoxin biosynthesis homologous genes (aflT, nor-i, aflR, norA, avnA, verB, and vbs), and group 2, having three homologous (avnA, verB, and vbs). Partial aflatoxin homologous gene cluster of RIB62 from group 2 was sequenced and compared with that of RIB40 from group 1. RIB62 showed a large deletion upstream of ver-1 with more than half of the aflatoxin homologous gene cluster missing including aflR, a positive transcriptional regulatory gene. Adjacent to the deletion of the aflatoxin homologous gene cluster, RIB62 has a unique sequence of about 8kb and a telomere. Southern analysis of A. oryzae RIB strains with four kinds of probe derived from the unique sequence of RIB62 showed that all group 2 strains have identical hybridizing signals. Polymerase chain reaction with specific primer set designed to amplify the junction between ver-1 and the unique sequence of RIB62 resulted in the same size of DNA fragment only from group 2 strains. Based on these results, we developed a useful genetic tool that distinguishes A. oryzae group 2 strains from the other groups' strains and propose that it might have differentiated from the ancestral strains due to chromosomal breakage.

• Journal of Microbiology and Biotechnology
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• v.34 no.8
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• pp.1671-1679
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• 2024

Aspergillus flavus, the primary mold that causes food spoilage, poses significant health and economic problems worldwide. Eliminating A. flavus growth is essential to ensure the safety of agricultural products, and extracellular compounds (ECCs) produced by Bacillus spp. have been demonstrated to inhibit the growth of this pathogen. In this study, we aimed to identify microorganisms efficient at inhibiting A. flavus growth and degrading aflatoxin B₁. We isolated microorganisms from soil samples using a culture medium containing coumarin (CM medium) as the sole carbon source. Of the 498 isolates grown on CM medium, only 132 bacterial strains were capable of inhibiting A. flavus growth. Isolate 3BS12-4, identified as Bacillus siamensis, exhibited the highest antifungal activity with an inhibition ratio of 43.10%, and was therefore selected for further studies. The inhibition of A. flavus by isolate 3BS12-4 was predominantly attributed to ECCs, with a minimum inhibitory concentration and minimum fungicidal concentration of 0.512 g/ml. SEM analysis revealed that the ECCs disrupted the mycelium of A. flavus. The hydrolytic enzyme activity of the ECCs was assessed by protease, β-1,3-glucanase, and chitinase activity. Our results demonstrate a remarkable 96.11% aflatoxin B1 degradation mediated by ECCs produced by isolate 3BS12-4. Furthermore, treatment with these compounds resulted in a significant 97.93% inhibition of A. flavus growth on peanut seeds. These findings collectively present B. siamensis 3BS12-4 as a promising tool for developing environmentally friendly products to manage aflatoxin-producing fungi and contribute to the enhancement of agricultural product safety and food security.

• Korean Journal of Food Science and Technology
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• v.21 no.3
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• pp.419-424
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• 1989

The effects of browning Products (BP) from Kanjang(soy sauce) and charcoal on the degradation of aflatoxin $B_1(AFB_1)$ in Kanjang and its model system were studied. Approximately 60% of $AFB_1$ was degraded in the presence of 0.05% BP at pH 7 of phosphate buffer after 2 days of incubation at $30^{\circ}C$. The mutagenicity of the $AFB_1$ which reacted with the BP was decreased to about 50% and 70% in Salmonella typhimurium TA98 and TA100 strains, respectively (p<0.05). When a few pieces of charcoal were added to home made Kanjang, $AFB_1$ was quite stable for 5days at $30^{\circ}C$, however, about 80% of $AFB_1$ was removed when the charcoal was either in distilled water or in 20% of NaCl solution after 2 days of incubation. Activated carbon instead of the charcoal removed $AFB_1$ completely in the all samples under the same conditions.

• Korean Journal of Agricultural Science
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• v.48 no.1
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• pp.73-81
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• 2021

The current study investigated how Saccharomyces cerevisiae ameliorates the adverse effects of aflatoxin on in vitro rumen fermentation. In this study, five groups (T₁: Control [basal feed]; T₂: T₁ + 300 ppb aflatoxin B1 [AFB₁] and T₃, T₄, and T₅: T₂ with 0.05, 0.1, and 0.2% of S. cerevisiae, respectively) were prepared and incubated in vitro. The results revealed that truly degradable dry matter (TDDM), gas production (GP), microbial biomass production (MBP), truly degradable organic matter (TDOM), partitioning factor (PF), total volatile fatty acids (TVFA), acetate (A), propionate (P) and butyrate (B) values in the control group (T₁) were higher (p < 0.05) than those of the AFB₁ fed group (T₂). The A : P ratio in the control group (T₁) was reduced (p < 0.05) when compared to that of the T₂ group. The TDDM, TDOM, GP, TVFA, A, P, and B values of T₃, T₄, and T₅ improved with the increasing levels of S. cerevisiae; however, the values of group T₅ were lower (p < 0.05) than that of the control. The values of MBP, A : P ratio and PF in group T₅ were statistically similar to that of the control. It was concluded that the inclusion of S. cerevisiae (0.05 to 0.20%) to the AFB₁ (300 ppb) contaminated feed partially to completely ameliorated the adverse effects of AFB₁ on the in vitro rumen fermentation parameters.

• Journal of Nutrition and Health
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• v.26 no.3
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• pp.268-276
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• 1993

Coffee is known to increase pancreatic secretion of digestive enzymes. The mutagen, aflatoxin B1(AFB1) is contained in fermented foods and known to increase the specific activities of pancreatic chymotrypsin, trypsi, amylase, and lipase. Nowadays, coffee intake is increased among Koreans who have consumed relatively high amount of traditional fermented foods. Therefore, this study was performed to examine the effect of coffee and AFB1 on pancreatic exocrine function and structure. Rats were divided into 10 experimental groups. The first five groups were W(control group), LD(0.2g decaffeinated coffee/Kg B.W), HD(3g decaffeinated coffee/Kg B.W), LC(0.2g coffee/Kg B.W), and HC(3g coffee/Kg B.W). The second five groups were WA, LDA, HDA, LCA, HCA, same as first five groups in caffieine level but treated with AFB1. The result of this experiment showed that the caffeine intake did not influence significantly on the growth and feed efficiency. But water intake was increased by caffeine intake and AFB1 treatment. The weights of pancreas and liver were increased as the caffeine intake was increased. Trypsin activities were tend to increase in concentrated coffee groups(HD, HC). AFB1 treated groups showed the higher trypsin level than the AFB1 untreated groups. Amylase activities were tend to increase in concentrated coffee groups(HD, HC) of AFB1 untreated animals. AFB1 treated did not show the additional effect on the stimulated amylase secretion by coffee. Lipase activities were tend to decrease in concentrated coffee groups(HD, HC) of AFB1 untreated animals. Lipase activities were increased in the order named WA group, coffee groups, decaffeinated coffee groups in AFB1 treated animals. AFB1 treated groups showed the higher lipase level than AFB1 untreated groups. In the histologic observation of pancreas HCA group showed more dense compound tubuloalveolar glands and proliferation of nuclei than normal. The result suggested a development of a atypia which is ongoing phase to a cancer.

• Food Science and Preservation
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• v.21 no.5
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• pp.747-756
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• 2014

The objective of this study was to examine the effects of vitamin C on the formation of aflatoxin $B_1$ ($AFB_1$)-DNA adduct and $AFB_1$-induing cellular oxidative damage in rat livers treated with radiation and $AFB_1$. Six-week-old male Sprague-Dawley rats were randomly divided into five groups: the control group, the $AFB_1$-treated group, the group treated with $AFB_1$ and vitamin C, the group treated with X-ray and $AFB_1$, and the group treated with X-ray and $AFB_1$ with vitamin C. On the first day of the experiment, only one dose of X-rays was exposed to the entire liver at 1,500 cGy. Next, vitamin C was injected at 10 mg/kg body weight via intraperitoneal injection, followed an hour later by the administration of 0.4 mg/kg of $AFB_1$ via intraperitoneal injection. These treatments were administered every three days for 15 days. On the 16th day, the animals were sacrificed. The $AFB_1$ contents of the rat sera were determined via indirect competitive ELISA. In the quantitative analysis of $AFB_1$ in the rat sera via ELISA, $5.17{\pm}0.34ng/mL$ of $AFB_1$ was detected in the $AFB_1$-treated groups, but the amount decreased more significantly to $3.23{\pm}0.76ng/mL$ in the groups treated with $AFB_1$ and vitamin C (p<0.01) than in the $AFB_1$-treated groups. The effect of vitamin C on $AFB_1$-DNA adduct formation was determined via ELISA. The values of $AFB_1$-DNA adduct formation were $9.38{\pm}0.41ng/mL$ in the $AFB_1$-treated groups, but the amount decreased more significantly to $5.28{\pm}0.32ng/mL$ in the groups treated with $AFB_1$ and vitamin C (p<0.01) than in the $AFB_1$-treated groups. Immunohistochemistry revealed that the accumulation of the $AFB_1$ was not observed in the normal liver tissue (G1). The $AFB_1$-positive materials were observed in the central vein and the portal vein of the liver tissue from the $AFB_1$(G2) treatment or the X-ray and $AFB_1$(G4) co-treatment, but the $AFB_1$-positive materials were observed weakly in the group treated with vitamin C (G3 and G5). These results indicate that vitamin C had ameliorating effects on the $AFB_1$ accumulation of liver tissue.

• Korean Journal of Food Science and Technology
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• v.32 no.6
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• pp.1258-1265
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• 2000

In oder to acquire microbial agents that can be utilized for control of aflatoxin produced by Aspergillus. flavus and Asp. parasiticus, antifungal bacteria were isolated. Antifungal bacteria was identified as Bacillus spp. based on morphology and physico-biochemical characteristics. Amount of aflatoxin $B_1$ from Doenjang(soybean paste) prepared with Asp. flavus, Asp. parasiticus, antifungal bacteria(Bacillus sp.), or mixture of Asp. flavus and Asp. parasiticus was 27.2 ppb, 30.3 ppb, 3.4 ppb, and 3.7 ppb, respectively. Aflatoxin $B_1$ was not detected from Doenjang(control) and Doenjang prepared with antifungal bacteria. Content and compositions of free sugars, fatty acid, organic acid and free amino acid in Doenjang prepared with Asp. flavus and Asp. parasiticus, antifungal bacteria and mixture of Asp. flavus and Asp. parasiticus were not significantly different. For volatile flavor compounds of Doenjang prepared with antifungal bacteria, 2-pentyl furan and butanoic acid were disappeared or reduced, while octadecene compounds were produced. However, those of Doenjang prepared with Asp. flavus or Asp. parasiticus and Doenjang(control) were not significantly different. These results suggested that the antifungal bacteria(Bacillus sp.) inhibited production of aflatoxin and that antifungal bacteria did not effect the quality of Doenjang.