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排序方式: 共有71条查询结果,搜索用时 15 毫秒
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Fungal contamination of agricultural commodities leads to their spoilage and renders them unfit for human consumption. Ergosterol, a predominant sterol in most fungi and a major constituent of the cell membrane, has been established as a reliable biochemical marker for fungal growth. Various chemical and physico‐chemical methods to quantify ergosterol as an index of fungal contamination are in practice. Yet, immunoassays are the methods of choice in food analysis due to their increased specificity, sensitivity and rapidity. This paper reports the synthesis of an antigen, bovine serum albumin‐ergosterol conjugate, and its immunocharacterization. Ergosterol was converted to ergosterol hemisuccinate (EHS) by succinylation. Subsequently, EHS was conjugated to bovine serum albumin by the mixed anhydride reaction. The molar ratio was found to be 1:28. Antisera raised against the synthesized antigen in rabbits was characterized by the Ouchterlony double‐diffusion technique and an antibody capture assay. Ouchterlony analysis showed a titre of 1:2. Further, characterization by an antibody capture assay, using 50 ng well (10 ng equivalent of ergosterol) of the antigen, gave a titre of 1:4000 dilution of antiserum, with an absorbance of 1.0 at 405 nm. The synthesized antigen may find an application in the development of an immunoanalytical method for ergosterol quantification as a measure of food quality in relation to fungal contamination. 相似文献
3.
Cloning,sequencing and analysis of the yeastS. uvarum ERG10 gene encoding acetoacetyl CoA thiolase 总被引:7,自引:0,他引:7
Sylvie Dequin Remi Gloeckler Christopher J. Herberte Françoise Boutelet 《Current genetics》1988,13(6):471-478
Summary TheERG10 gene specific toS. uvarum, a brewing yeast, has been cloned by complementation of anS. cerevisiae erg10 mutant.S. uvarum contains two differentERG10 genes. One of these is similar to theS. cerevisiae ERG10 gene; they are structurally different, but functionally homologous. The clonedERG10 gene has been located on chromosome XVI, and we have shown that it is allelic to the previously isolatedtsm0115 mutants. Northern blot and sequence analysis indicate that theERG10 gene is highly expressed, and biochemical and genetic evidence show that it encodes the cytoplasmic acetoacetyl CoA thiolase. 相似文献
4.
Fabian F Moebius Raphael J Reiter Markus Hanner Hartmut Glossmann 《British journal of pharmacology》1997,121(1):1-6
- The sigma-drug binding site of guinea-pig liver is carried by a protein which shares significant amino acid sequence similarities with the yeast sterol C8–C7 isomerase (ERG2 protein). Pharmacologically - but not structurally - the sigma1-site is also related to the emopamil binding protein, the mammalian sterol C8–C7 isomerase. We therefore investigated if sterol C8–C7 isomerase inhibitors are high affinity ligands for the (+)-[3H]-pentazocine labelled sigma1-binding site.
- Among the compounds which bound with high affinity to native hepatic and cerebral as well as to yeast expressed sigma1-binding sites were the agricultural fungicide fenpropimorph (Ki 0.005 nM), the antihypocholesterinaemic drugs triparanol (Ki 7.0 nM), AY-9944 (Ki 0.46 nM) and MDL28,815 (Ki 0.16 nM), the enantiomers of the ovulation inducer clomiphene (Ki 5.5 and 12 nM, respectively) and the antioestrogene tamoxifen (Ki 26 nM).
- Except for tamoxifen these affinities are essentially identical with those for the [3H]-ifenprodil labelled sterol C8–C7 isomerase of S. cerevisiae. This demonstrates that sigma1-binding protein and yeast isomerase are not only structurally but also pharmacologically related. Because of its affiliations with yeast and mammalian sterol isomerases we propose that the sigma1-binding site is localized on a sterol isomerase related protein, involved in postsqualene sterol biosynthesis.
5.
John P. R. Keon Caroline S. James Samantha Court Catharine Baden-Daintree Andrew M. Bailey Raymond S. Burden Martin Bard John A. Hargreaves 《Current genetics》1994,25(6):531-537
The Magnaporthe grisea ERG2 gene, encoding 87 sterol isomerase, was isolated from a genomic library by heterologous hybridization to a fragment of the Ustilago maydis ERG2 gene. The isolated gene contained a reading frame of 745 bp which encoded a protein of 221 amino acids. The coding region was interrupted by a single putative 79-bp-long intron. The deduced amino-acid sequence exhibited similarity to the ERG2 gene products of U. maydis and of Saccharomyces cerevisiae, particularly in the central region of the proteins. The NH2-terminal of all three proteins contained a long stretch of amino acids that were strongly hydrophobic, suggesting that they may function by anchoring the protein to a membrane surface. The M. grisea ERG2 gene complemented a U. maydis deletion mutant in which the ERG2 gene had been removed using a one-step gene replacement procedure. The 87 sterol isomerase produced by the M. grisea ERG2 gene exhibited a level of sensitivity to the sterol biosynthesis inhibitor, tridemorph, similar to that of the enzyme derived from the U. maydis ERG2 gene. 相似文献
6.
目的:分别采用高效液相色谱法和气相色谱质谱联用法测定白念珠菌麦角甾醇含量,并对这两种方法进行比较。方法:采用浓度梯度递增法诱导白念珠菌耐药株,采用KONT真菌显色MIC药敏系统鉴定耐药性。构建6、12、24h生物被膜,采用高效液相色谱法和气相色谱质谱联用法测定麦角甾醇含量。结果:高效液相色谱法检测出麦角甾醇含量,最低检出浓度为0.05mg/L,而气相色谱质谱联用法未检测出麦角甾醇含量。结论:高效液相色谱法能准确测定白念珠菌麦角甾醇含量。与气相色谱质谱联用法比较,高效液相色谱法简单、高效和可靠。 相似文献
7.
目的比较虎乳灵芝栽培品和野生品脂溶性化学成分的差异。方法采用回流提取法提取虎乳灵芝野生品和栽培品(包括大米和木屑培养)菌核的脂溶性成分,用气相色谱-质谱联用(GC-MS)法分析栽培品和野生品的化学成分,并计算各化学成分的相对百分含量。结果虎乳灵芝大米、木屑栽培品及野生品分别鉴定出11种、10种、14种化合物。主要为烃类化合物,三者共有成分为2,4-二叔丁基苯酚、二十五烷和三十一烷等,并且二十五烷和三十一烷在大米栽培品、木屑栽培品和野生品中相对百分含量分别为15.85%、17.36%、16.43%和12.01%、11.58%、19.29%。三者成分差异表现为:大米栽培品含有麦角甾醇、二十九烷;木屑栽培品含有二十二烷、二十三烷、二十四烷;野生品还含有二十四烷、芥酸酰胺、邻苯二甲酸二丁酯、亚油酸乙酯。结论栽培品和野生品的脂溶性成分中含有2,4-二叔丁基苯酚、二十五烷和三十一烷等相同的化学成分,但又存在明显的差异,可为虎乳灵芝栽培品的质量评价和开发利用提供参考。 相似文献
8.
《Journal de Mycologie Médicale》2020,30(1):100921
AimAmong the most common mechanisms of multidrug resistance (MDR) in prevalent human fungal pathogen, Candida albicans, overexpression of drug efflux pumps remains the predominant mechanism. Hence to inhibit efflux pumps and chemosensitize C. albicans against traditional antifungal drugs still represents an attractive approach. The present study aimed to analyze the effect of Vanillin (Van), a natural food flavoring agent, on drug efflux pump activity of Candida albicans.Methods and resultsWe observed that Van specifically inhibits Candida drug resistance protein 2 (CaCdr2p) activity belonging to ATP Binding Cassette (ABC) superfamily as revealed by abrogated rhodamine 6G efflux and nile red accumulation assay with CaCdr2p over expressing strain. Insight studies into the mechanisms suggested that abrogated efflux by CaCdr2p is due to competitive mode of inhibition by Van as depicted by Lineweaver-Burk plot. RT-PCR, western blot and confocal microscopy further unraveled that Van leads to reduced expression of CDR2 and CaCdr2p mislocalization respectively. Furthermore, Van sensitizes the azole sensitive and resistant clinical matched pair of isolates Gu4/Gu5 and led to abrogated rhodamine 6G efflux and depleted ergosterol. Furthermore, Van synergizes with membrane targeting drugs fluconazole and amphotericin B as their fractional inhibitory coefficient index was less than 0.5.ConclusionVan being a potent inhibitor of CaCdr2p and chemosensitizing of drug resistant C. albicans warrants further studies to be exploited as effective antifungal agent. 相似文献
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10.
Hippelein M Rügamer M 《International journal of hygiene and environmental health》2004,207(4):379-385
Ergosterol was used as a specific indicator of fungal biomass to determine and assess mould growth on damp building material. The samples were saponified, cleaned up on a silica gel column and the sterols silylated and analysed by gas chromatography and mass spectrometry in MS/MS mode. Ninety-one samples of building materials from damp rooms were analysed including plaster, plaster/paint, paint, wood-chip wallpaper and paper wall covering. The concentrations of ergosterol ranged from 0.1 to 130 microg/g dry mass and depended on carbon content of the material. The highest concentrations were determined for wallpaper, the lowest for plaster and intermediate ones for paint. Based on ergosterol content and inspection of the room during sampling a rough classification of mould infestation is presented. The applicability of the ergosterol method was further tested in two case studies on the spatial distribution of fungi on damp walls and irregular distributions were found. With few exceptions the concentration of ergosterol in building materials was found to be a suitable indicator to estimate fungal biomass. 相似文献