Immunochemical and mass spectrometric analysis of Nε-(carboxymethyl)lysine content of AGE-BSA systems prepared with and without selected antiglycation agents

The present study was designed to compare surface plasmon resonance (SPR) biosensor, enzyme-linked immunosorbent assay (ELISA), and ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) methods for the analysis of Nε-(carboxymethyl)lysine (CML) in glucose-bovine serum albumin (BSA) model systems and to investigate the possible inhibitory effect of selected compounds (α-tocopherol, ferulic acid, rutin, thiamin, thiamin monophosphate, and thiamin pyrophosphate) on CML formation. The reported levels of CML detected were dependent upon the method of analysis employed. The highest reported concentrations were obtained with the SPR biosensor, whereas the lowest were found by ELISA. However, a high correlation was observed between these two immunochemical procedures. CML concentrations were dependent upon the type and concentration of the candidate CML inhibitor. All inhibitory compounds investigated, with the exception of α-tocopherol, decreased the level of CML formation in the glucose-BSA system.     

Effect of cooking on concentrations of β-estradiol and metabolites in model matrices and beef

Because beef food products are generally cooked prior to consumption, the behavior of chemicals in these cooked foods is important in estimating human exposure. The heat stability of the natural estrogen β-estradiol (β-E2) and its metabolites α-estradiol (α-E2), estrone (E1), and several catechol estrogens was examined in heated vegetable oil and aqueous solutions. The chemicals were also incorporated into regular and extra lean ground beef and subjected to cooking. E1 and E2 were stable in aqueous solutions at 100°C, whereas the catechol estrogens exhibited first-order decay curves with half-lives of 2-10 min. Their stability improved to the same level as the other test chemicals when an antioxidant was added to the solution, suggesting that their disappearance was due to oxidation rather than thermal degradation. E1 and E2 were also stable in heated vegetable oil (160-180°C), whereas catechol estrogen decreased 30-50% over the 2 h duration of the experiments. Chemical losses from cooked beef appear to be related to the fat content of the beef, with greater losses occurring in regular ground beef (25-30%), compared to extra lean ground beef (5-20%). This study shows that cooking reduces but does not eliminate the potential for dietary exposure to growth promoters in ground beef.     

Effect of 2(N‐morpholino)ethane)sulfonic acid (MES) on the growth and tissue composition of cucumber

The effect of 2(N‐morpholino)ethane)sulfonic acid (MES) on the growth of cucumber (Cucumis sativa L. var. Marketer) in hydroponic culture was determined at 0, 1, 5, and 10 mM concentrations. The effect of adding the MES to the solution at the time of transfer to hydroponic culture or waiting one week was also determined. MES was observed to strongly affect plant growth with increasing concentration in nutrient solution. Tissue and nutrient solution analysis determined that MES affects manganese (Mn) uptake. MES appears to be reduced by Mn, precipitating the Mn out of solution. The suitability of MES as a pH buffer in hydroponic culture is discussed in terms of this effect.     

Effect of B,Mn and Zn on nodulation and N2‐fixation in southernpeas

Abstract

Two greenhouse studies were conducted to evaluate the effect of B, Mn and Zn on nodulation and N₂‐fixation of southernpea (Vigna unguiculata (L.) Halp.) cultivars ‘Freezegreen’, ‘Mississippi Silver’ and ‘Pinkeye Purple Hull’. The cultivars were grown in plastic pots with a Norfolk sandy loam (fine, loamy siliceous thermic, Typic Paleudult) soil treated with B, Mn and Zn at rates of 0, 5, 10 and 20 kg/ha each at pH levels 5.5, 6.0 and 6.5. At pH 6.5 all micronutrient treatments significantly increased nodulation and N₂‐fixation over the control (no micronutrient applied). The effects of B, Mn and Zn on nodulation and N₂‐fixation depended on the cultivar and soil pH. For plants given the 5 kg/ha B and Mn treatments, ‘Mississippi Silver’ produced the highest number of nodules and ‘Pinkeye Purple Hull’ the least. At 20 kg/ha Zn, nodulation of ‘Freezegreen’ was highest and ‘Pinkeye Purple Hull’ the lowest. As a whole, maximum nodulation was at 5 kg/ha B and Mn and 20 kg/ha for Zn. Nitrogen fixation rates responded similarly except that the optimum rate for Zn was 10 kg/ha. Seed yield of plants peaked at 5 kg/ha for B and 10 kg/ha for Zn, indicating a possible relation of N₂‐fixation to seed yield.     

Effect of N‐form on macronutrient and micronutrient concentration and uptake of creeping bentgrass 1

Abstract

Nitrogen‐form effect on nutrient uptake and the subsequent concentration of nutrients in turfgrass plant tissue has not been thoroughly investigated. This study evaluated the effects of clipping regime and N‐form on the tissue concentration of macronutrients and micronutrients and macronutrient uptake in ‘Penncross’ creeping bentgrass (Agrostis palustris Huds.). Turfgrass plugs were grown under greenhouse conditions in a modified Hoagland's solution with a combination of three nutrient solutions (100% NO₃ ^?, 100% NH₄ ⁺, and 50:50 ratio of NH₄ ⁺:NO₃ ^?) and two cutting regimes (cut and uncut). Concentrations of macronutrients and micronutrients were determined for shoot, root and verdure. Nutrient uptake was determined weekly. Uncut NO₃ ^?‐treated plants accumulated higher concentrations of K, Ca, Mg, B and Cu in the shoot tissue; P, K, Ca, Mg, B, Cu, Mn and Zn in the root tissue; and P, Ca, Mg, B, Fe and Mn in the verdure compared to uncut NN₄ ⁺‐treated plants. Nitrate uptake was greater with uncut NO₃ ^?‐treated plants than was NH₄ ⁺ absorption with uncut NH₄ ⁺‐treated plants. Plants grown with the uncut 50:50 treatment adsorbed more NH₄ ⁺ than NO₃ ^?. Plants grown with the uncut NO₃ ^? and 50:50 treatments adsorbed higher amounts of P, K, and Ca compared to the NH₄ ⁺ treatment. The cut NO₃ ^?‐treated plants accumulated higher concentrations of K in the shoot tissue; P, Ca, Mg, B, Cu, Fe and Mn in the root tissue; and B in the verdure than did the cut NH₄ ⁺‐treated plants. Cut NO₃ ^?‐treated plants adsorbed less NO₃ ^? than did cut NH₄ ⁺‐treated plants adsorbed NH₄ ⁺. The cut 50:50 treatment adsorbed more NH₄ ⁺ than NO₃ ^?. Plants grown with NO₃ ^? and 50:50 treatments, under both cutting regimes, resulted in higher concentrations of most macro‐ and micronutrients and greater nutrient uptake compared to the NH₄ ⁺‐treated plants.     

Effect of N‐form on growth and nutrient content of creeping bentgrass 1

Abstract

The primary nitrogen forms utilized by plants are ammonium and nitrate. Although the importance of nutrients other than nitrogen for proper turfgrass growth is well established, the amounts of these nutrients in the plant tissue in relation to the use of different N‐forms has not been clearly documented. This study was conducted under greenhouse conditions to determine the effect of N‐form and cutting regime on growth, macronutrient, and micronutrient content of creeping bentgrass (Agrostis palustris Huds. ‘Penncross'). Treatments consisted of 100% NO3^? (calcium nitrate), 100% NH₄ ⁺ (ammonium sulfate), and a 50:50 ratio of NH₄ ⁺:NO₃ ^?. Half the turfgrass plants were maintained at a height of 1 cm (cut), while the other half of the plants were not cut until the end of the study (uncut). The uncut 50:50 treatment yielded the highest shoot, verdure, and total plant dry matter, while the uncut NO₃ ^? treatment produced the highest root dry matter. The uncut NH₄ ⁺ treatment yielded the least shoot, root, and total plant dry matter. Plants of the uncut NO₃ ^? treatment had greater accumulation of macronutrients in the shoot and root tissue compared to plants of the NH₄ ⁺ treatment. The uncut NO₃ ^? and 50:50 treatments had higher total accumulation of micronutrients compared to the uncut NH₄ ⁺‐treated plants. The cut NO₃ ^? treatment resulted in the highest macronutrient and micronutrient contents in the root tissue in comparison to other cut treatments. The cut treatments had the highest percentage accumulation of nutrients in the verdure tissue, while the uncut treatments had the highest percentage accumulation of nutrients in the shoot tissue.     

Effect of Synergism and Antagonism between Metals on Toxicity in Soils

Synergism and antagonism of cadmium(Cd),copper (Cu) and selenium (Se) to biological toxicities in red soil,yellow brown soil and black soil were evaluated by MICROTOX method.The relation between forms of the tested metals in soil and the synergism or antagonism between them was also studied.Results showed that owing to the difference of soil chemical properties,toxicity of these metals in soils was different.In red soil with acid reaction and low in cation exchange capacity,antagonism occurred significantly between metals when they coexisted at high concentrations,while synergism occurred only under low concentrations.It is indicated that in red soil,toxicity of metals affected by synergism or antagonism depends on concentration of the metals present.For yellow brown soil and black soil with larger cation exchange capacity and lower exchangeable aluminium(Al),no toxicity of metals was observed even if metals were added to soil in high concentrations.Synergism and antagonism between Cd,Cu and Se were controlled by the forms of metals present.The amount of water-soluble metals was the most important factor in determining synergism and antagonism. In this paper,comparisons of synergism and antagkonism between metals in soils and in water solutions were made.There occurred the synergism of metal toxicity in water solutions when the concentration of coexisting metals was high.This is just opposite to the case in soils.     

Effect of cycloheximide on N2O and NO3– production in a forest and an agricultural soil

The present work aims at evaluating the effect of cycloheximide at concentrations of between 0.5 and 5mgg^–1 on N₂O and NO₃ ^– production in two slightly alkaline soils, sampled from deciduous woodland and arable cultivation. In the first experiment, peptone was used as the “inducing substrate” for heterotrophic activity, and soil was incubated with cycloheximide (at different concentrations) and/or acetylene (1mll^–1) to block induced eukaryotic protein synthesis and ammonia monooxygenase activity, respectively. Peptone addition stimulated N₂O and NO₃ ^– production significantly in woodland soil, whereas arable soil showed no significant N₂O emissions and low NO₃ ^– production. Low cycloheximide concentrations drastically reduced N₂O emissions in woodland soil, suggesting a potential role of fungi in N₂O emissions. However, acetylene was equally effective in blocking N₂O emissions and part of NO₃ ^– production, so that a possible role of ammonia monooxygenase in an organic-inorganic pathway of N nitrification in fungal metabolism can be hypothesized. A second experiment was carried out on the woodland soil to check if low cycloheximide concentrations had non-target biocidal effects on soil microorganisms. Attention was focused on the range of concentrations which had reduced N₂O emission in the woodland soil. The results suggested that at concentrations of cycloheximide between 0.5 and 2mgg^–1 any biocidal effect on microbial biomass was negligible in the first 48h; therefore only selective inhibition of protein synthesis could be expected. The whole nitrifier population seemed to be particularly sensitive to cycloheximide concentrations higher than 2.5mgg^–1. Received: 4 July 1997     

Effect of N‐(n‐butyl)thiophosphoric triamide added to peat and leather in urea‐based fertilizers on urea hydrolysis and ammonia volatilization

Abstract

A laboratory experiment evaluated the rate of urea hydrolysis and ammonia volatilization from urea (U) mixed in organo‐mineral (O‐M) fertilizers. These fertilizers were incubated in soil in the presence or absence of N‐(n‐butyl)thiophosphoric triamide (NBPT) as a urease inhibitor. Two organic matrices, leather (L) and peat (P), were used to prepare the O‐M fertilizers. In the absence of NBPT, the highest ammonia losses and the fastest rate of urea hydrolysis were in the soil treated with the fertilizer containing leather (UL₅₀). Significantly lower ammonia losses occurred with peat‐based fertilizers. Although the fertilizer containing peat (UP₅₀) stimulated the rate of urea hydrolysis with respect to the urea alone, no increase in ammonia volatilization was detected. NBPT‐containing fertilizers were stored for different times (0,7, 30, and 60 days) and temperatures (25°C and 40°C), and the NBPT recovery was monitored by extraction and analysis by HPLC. The NBPT recovery decreased by increasing either the storage time or the storage temperature. Differences among the fertilizers occurred after storage at 40°C for 30 or 60 days. With UN, in spite of about 25% extracted amount of NBPT, the ammonia losses did not increase with respect to the non‐stored fertilizer. On the contrary, no inhibitor was recovered from either of the O‐M fertilizers (UNL and UNP). However, in the presence of leather, NBPT reduced the volatilization losses by 35 to 40%, whereas in the presence of peat, a complete loss of NBPT efficiency occurred. In general, either the inhibitor recovery or efficiency were affected by the storage conditions or the type of organic matrix.     

Effect of dicyandiamide on the form and recovery of 15N‐labeled urea in the delayed flood soil system

Abstract

Dicyandiamide (DCD) is a nitrification inhibitor that has been proposed for use in drill‐seeded rice. Immobilization of fertilizer NH₄ ⁺‐N by soil microorganisms under aerobic conditions has been found to be significantly enhanced in the presence of a nitrification inhibitor. The objective of this laboratory study was to determine if DCD significantly delayed nitrification of urea‐derived N, and if this enhanced immobilization of the fertilizer N in the delayed‐flood soil system inherent to dry‐seeded rice culture. Nitrogen‐15‐labeled urea solution, with and without DCD (1: 9 w/w N basis), was applied to a Crowley silt loam (Typic Albaqualf) and the soil was incubated for 10 weeks in the laboratory. The soil was maintained under nonflooded conditions for the first four weeks and then a flood was applied and maintained for the remaining six weeks of incubation. The use of DCD significantly slowed the nitrification of the fertilizer N during the four weeks of nonflooded incubation to cause the (urea + DCD)‐amended soil to have a 2.5 times higher fertilizer‐derived exchangeable NH₄+‐N concentration by the end of the fourth week. However, the higher exchangeable NH₄+‐N concentration had no significant effect on the amount of fertilizer N immobilized during this period. Immobilization of the fertilizer N appeared to level off during the nonflood period about the second week after application. After flooding, immobilization of fertilizer N resumed and was much greater in the (urea + DCD)‐amended soil that had the much higher fertilizer‐derived exchangeable NH₄ ⁺‐N concentration. Immobilization of fertilizer N appeared to obtain a maximum in the urea‐amended soil (18%) about two weeks after flooding and for the (urea + DCD)‐amended soil (28%) about four weeks after flooding.     

Impact of high-pressure processing on vitamin E (α-, γ-, and δ-tocopherol), vitamin D (cholecalciferol and ergocalciferol), and fatty acid profiles in liquid foods

In the present study, four high-pressure (HP) treatments (100, 200, 300, and 400 MPa) of 9 min duration were evaluated to assess their effect on the lipid fraction (fat-soluble vitamins and fatty acid profile) of an orange juice-milk and a vegetable beverage. After HP treatment, nonsignificant changes in vitamin D(2) and D(3) contents were observed for both beverages. An increase in vitamin E activity was observed in HP beverages when pressures >100 MPa were applied, mainly due to an increase in α-tocopherol content. Only a small reduction in fat content was found for the orange juice-milk beverage, but no changes were observed for the vegetable beverage. A significant decrease in SFA levels was observed in HP-treated (300-400 MPa) orange juice-milk. With regard to MUFA, a significant increase in oleic acid (C(18:1)) was found in both liquid foods. Nonsignificant differences in the PUFA profiles were observed after HP processing.     

Effect of hydroxypropyl-β-cyclodextrin on diuron desorption and mineralisation in soils

Purpose

The herbicide diuron has the unfortunate property of being strongly adsorbed onto soil organic matter particles, and hence, is slowly degraded in the environment because of its reduced bioavailability. The aim of this work was to gain insight into the fate and behaviour of diuron in the soil–water system, and develop and test an environmentally friendly soil decontamination technique that could give rise to an enhancement of diuron mineralisation by sensitive soil endogenous microorganisms, by means of increasing the bioavailability of the pollutant employing cyclodextrin (CD) solutions what would represent an improvement from both economic and environmental standpoints.

Materials and methods

Selected soil colloidal components: montmorillonite, a synthetic humic acid and a synthetic acicular goethite, and two different soils were employed in this study to perform batch adsorption–desorption experiments. Desorption experiments were performed using a 0.01 M Ca(NO₃)₂ solution with and without hydroxypropyl-β-cyclodextrin (HPBCD) 50 mM. Assays to study the mineralisation of ¹⁴C-labelled diuron were performed in respirometers, into which 10 g of soil and 50 mL of mineral salts medium (MMK) were placed, obtaining a final concentration of 50 mg?kg^?1 and a radioactivity of approximately 900 Bq per flask.

Results and discussion

Humic acid could be clearly discerned as the major colloidal component responsible for adsorption. HPBCD was used in diuron desorption experiments from soil, showing a strong extracting power on its removal. The mineralisation of diuron in the presence and absence of HPBCD was tested in a soil managed with diuron for several years, involving therefore the presence of microorganisms that have some specificity for diuron. Natural soil attenuation for diuron was improved when a HPBCD solution was used in the presence of micronutrients as a bioavailability enhancer, obtaining 66 % of mineralisation in comparison to that obtained with only micronutrients addition (44 %).

Conclusions

The use of HPBCD solution at a very low concentration of only 10 times the diuron equimolar concentration in soil, acts as a bioavailability enhancer, accelerating the passage of the diuron-desorbing fraction from the soil particle surface to the soil solution, and hence, improving the accessibility of the microorganisms to the herbicide. Diuron mineralisation rate and the extent of its mineralisation were improved when the HPBCD solution was employed in the presence of micronutrients.     

Formation of arginine modifications in a model system of Nα-tert-butoxycarbonyl (Boc)-arginine with methylglyoxal

The present study deals with the mechanistic reaction pathway of the α-dicarbonyl compound methylglyoxal with the guanidino group of arginine. Eight products were formed from the reaction of methylglyoxal with N(α)-tert-butoxycarbonyl (Boc)-arginine under physiological conditions (pH 7.4 and 37 °C). Isolation and purification of substances were achieved using cation-exchange chromatography and preparative high-performance liquid chromatography (HPLC). Structures were verified by nuclear magnetic resonance (NMR) and high-resolution mass spectrometry. 2-Amino-5-(2-amino-4-hydro-4-methyl-5-imidazolinone-1-yl)pentanoic acid (3) was determined as the key intermediate precursor within the total reaction scheme. Kinetic studies identified N(δ)-(5-methyl-4-oxo-5-hydroimidazolinone-2-yl)-L-ornithine and N(7)-carboxyethylarginine as thermodynamically more stable products from compound 3. Further mechanistic investigations revealed an acidic hydrogen at C-8 of compound 3 to trigger aldol condensations. This reactivity of compound 3 allowed for the addition of another molecule of methylglyoxal to form products, such as N(δ)-(4-carboxy-4,6-dimethyl-5,6-dihydroxy-1,4,5,6-tetrahydropyrimidine-2-yl)-l-ornithine and argpyrimidine.     

Effect of Straw Incorporation on 15N‐Labeled Ammonium Nitrogen Uptake and Rice Growth

Abstract

A greenhouse experiment was conducted to determine the effect of rice straw residue on growth and uptake of added ¹⁵N‐labeled ammonium nitrogen (NH₄‐N) (3% ¹⁵N abundance at the rate of 150 kg N ha^?) by rice in Crowley silt loam soil (Typic Albaqualfs). Higher rates of rice straw addition had an adverse affect on plant growth from the first to sixth week. After 6 weeks, the high rice straw treatment had a positive effect on plant growth (P<0.05). The ¹⁵N‐labeled ammonium or fertilizer nitrogen (N) uptake by rice was significantly lower (P<0.05) in the high rice straw treatment as compared to lower rice straw treatments. Greater plant growth was recorded under alternate flooding and draining condition as compared to continuously flooded treatment (P<0.01).     

Effect of MES [2(N‐morpholino)ethanesulfonic acid] and amberlite IRC‐50 resin on nutrient pH control and soybean growth

The relative effectiveness of Amberlite IRC‐50 resin, in a recirculating ion exchange column, and several concentrations of MES [2(N‐mor‐pholino)ethanesulfonic acid] were evaluated for control of nutrient solution pH of hydroponically cultured soybean (Glycine max [L.] Merr. cv Williams). A low buffering capacity, urea‐based nutrient solution was used. MES buffer, at the concentrations evaluated (1,2, 4, and 10 mM), was less effective at maintaining desired pH than the IRC‐50 resin system. The pH of the 4.0 mM MES buffered nutrient solution decreased from 6.5 to below 4.0 in five days with 25 to 30 day old plants. By comparison, the pH of the IRC‐50 resin buffered nutrient solution dropped from 6.5 to 5.9 during the same period. Nitrate uptake by 27 to 29 day‐old plants was significantly slower when solutions were buffered with MES than with IRC‐50 resin. Rate of nitrate uptake decreased with increasing MES buffer concentrations and decreasing pH in short‐term studies on plants previously grown on IRC‐50 resin buffered nutrient solution. Mass of the plants grown on IRC‐50 resin buffered nutrient solution equaled or exceeded that of plants grown on 1, 2, or 4 mM MES. Total elemental uptake (mg/plant) by plants grown on nutrient solution buffered by IRC‐50 resin, for the ten elements tested, was equal to or greater than uptake by plants grown on nutrient solution buffered by 1, 2, or 4 mM MES. It is concluded that IRC‐50 resin in recirculating columns provides better pH control than does MES buffer for hydroponically grown soybean.     

Effect of zinc levels on phosphorous and zinc content in sand‐cultured soybeans

Soybeans (Glvcine max L.) cv. “Clark”; were grown in the greenhouse in sand‐filled plastic pots sub‐irrigated with Hoagland No. 1 solution to determine the possible inhibitory effects of Zn on the uptake of P. Zinc rates used were 0.05 (control), 0.25, 0.5 and 2.5 ppm. Yields equalled the control at Zn levels of 0.25 and 0.5 ppm, but plants grown in solutions containing 2.5 ppm Zn were stunted severely. Foliar Zn and P levels differed very little among Zn treatments. Zinc levels were highest and P levels were lowest, however, in the roots and stems of soybeans grown in solutions of 2.5 ppm Zn. Zinc and P uptake was significantly inhibited in the leaves, stems and roots of plants grown at the highest Zn rate. The recycling of wastes containing high Zn content could adversely affect plant growth by a suspected antagonism with P.     

Cationic antimicrobial (ε-polylysine)-anionic polysaccharide (pectin) interactions: influence of polymer charge on physical stability and antimicrobial efficacy

The cationic biopolymer ε-polylysine (ε-PL) is a potent food-grade antimicrobial that is highly effective against a range of food pathogens and spoilage organisms. In compositionally complex systems such as foods and beverages, cationic ε-PL molecules may associate with anionic substances, leading to increased turbidity, sediment formation, and reduced antimicrobial activity. This study therefore characterized the interactions between cationic ε-PL and anionic pectins with different degrees of esterification (DE) and then investigated the influence of these interactions on the antimicrobial efficacy of ε-PL. The nature of the interactions was characterized using isothermal titration calorimetry (ITC), microelectrophoresis (ME), and turbidity measurements. High (DE 61%), medium (DE 51%), and low (DE 42%) methoxyl pectins interacted with ε-PL molecules through electrostatic forces, forming either soluble or insoluble complexes with various electrical charges, depending on the relative mass ratio of pectin and ε-PL. The interaction of pectin with ε-PL increased as the negative charge density on the pectin molecules increased, that is, with decreasing DE. The antimicrobial efficacy of ε-PL against two acid-resistant spoilage yeasts (Zygosaccharomyces bailii and Saccharomyces cerevisiae) decreased progressively in the presence of increasing levels of all three pectins. Nevertheless, the low DE pectin decreased the antimicrobial efficacy of ε-PL much more dramatically, likely due to strong electrostatic binding of ε-PL onto low DE pectin molecules reducing its interaction with anionic microbe surfaces. This study provides knowledge that will facilitate the rational application of ε-PL as an antimicrobial in complex food systems.     

Homology modeling and screening of new 14α-demethylase inhibitor (DMI) fungicides based on optimized expression of CYP51 from Ustilago maydis in Escherichia coli

Ustilago maydis infection is a serious disease affecting corn crops worldwide. Sterol 14α-demethylase (CYP51) is one of the key enzymes of sterol biosynthesis and an effective target of antifungal drugs. To further study the interaction between CYP51 and drugs and exploit more specific 14α-demethylase inhibitor (DMI) fungicides for U. maydis, in this study homology modeling of CYP51 from U. maydis (UmCYP51) templated as the eukaryotic orthologues (the human CYP51) and screening of new DMI fungicides based on optimized expression were carried out for the first time. In addition, XF-113 and ZST-4 were screened by analyzing the spectral characteristics between the purified UmCYP51-35 and fungicides. These results provide a theoretical basis and new ideas for efficient design and development of new antifungal drugs.