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鉴定与棒曲霉素转化为表曲霉素过程相关的两种脱氢酶。

Characterization of Two Dehydrogenases from Involved in the Transformation of Patulin to Ascladiol.

机构信息

Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada.

Guelph Research and Development Centre, Agriculture and Agriculture-Food Canada, Guelph, ON N1G 5C9, Canada.

出版信息

Toxins (Basel). 2022 Jun 21;14(7):423. doi: 10.3390/toxins14070423.

Abstract

Patulin is a mycotoxin that primarily contaminate apples and apple products. Whole cell or cell-free extracts of ATCC 621 were able to transform patulin to E-ascladiol. Proteins from cell-free extracts were separated by anion exchange chromatography and fractions with patulin transformation activity were subjected to peptide mass fingerprinting, enabling the identification of two NADPH dependent short chain dehydrogenases, GOX0525 and GOX1899, with the requisite activity. The genes encoding these enzymes were expressed in and purified. Kinetic parameters for patulin reduction, as well as pH profiles and thermostability were established to provide further insight on the potential application of these enzymes for patulin detoxification.

摘要

展青霉素是一种主要污染苹果和苹果产品的真菌毒素。ATCC 621 的全细胞或无细胞提取物能够将展青霉素转化为 E-表没食子儿茶素。无细胞提取物中的蛋白质通过阴离子交换色谱分离,具有展青霉素转化活性的级分进行肽质量指纹图谱分析,鉴定出具有所需活性的两种 NADPH 依赖性短链脱氢酶,GOX0525 和 GOX1899。这些酶的编码基因在大肠杆菌中表达并纯化。建立了展青霉素还原的动力学参数以及 pH 曲线和热稳定性,为这些酶在展青霉素解毒中的潜在应用提供了进一步的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f29a/9323132/36915388d8b2/toxins-14-00423-g001.jpg

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