Alcohol oxidase (AOX) is an important flavin adenine dinucleotide (FAD) dependent oxidoreductase, which is responsible for converting methanol into formaldehyde and hydrogen peroxide for the growth of methylotrophic yeast Candida boidinii. Although AOX plays a crucial role in methanol catabolism, the experimental structure of AOX from Candida boidinii has not been elucidated. This study reports the first complete in silico model of AOX from C. boidinii. This paper also reports the AOX structure modeled using the threading approach, followed by structure analysis and molecular dynamics simulation. The modeled structure was compared with the aryl alcohol oxidase structure (a glucose–methanol–choline family member, pdbID: 3fim). A docking study was performed to analyze the interaction between AOX and its cofactor FAD. The AOX modeled structure also exhibited high similarity with respect to the FAD binding sites, which are the substrate binding sites as seen with 3fim. It was observed that the adenosine part of FAD was deeply buried inside AOX while the isoalloxazine ring stuck to the surface. This paper reports the interaction of selective proton ionophores (CCCP and DNP) with AOX and also reports their binding sites. These proton ionophores showed competitive binding with FAD. The occupancy of the FAD binding sites by the proton ionophore may lead to blocking of the entry of FAD and thereby disruption of AOX import into peroxisomes.

中文翻译：

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博伊丁氏假丝酵母醇氧化酶的建模及质子离子载体与FAD与酶的竞争结合的计算机分析

酒精氧化酶（AOX）是一种重要的黄素腺嘌呤二核苷酸（FAD）依赖性氧化还原酶，负责将甲醇转化为甲醛和过氧化氢，以促进甲基营养型博伊丁氏假丝酵母的生长。尽管AOX在甲醇分解代谢中起着关键作用，但尚未阐明Bodinii假丝酵母AOX的实验结构。这项研究报告了第一个完整的Boidinii C的AOX的计算机模拟模型。本文还报告了使用穿线方法对AOX结构进行建模，然后进行结构分析和分子动力学模拟。将模拟的结构与芳基醇氧化酶结构（葡萄糖-甲醇-胆碱家族成员，pdbID：3fim）进行了比较。进行了对接研究以分析AOX及其辅因子FAD之间的相互作用。AOX建模的结构相对于FAD结合位点也显示出高度相似性，FAD结合位点是3fim所见的底物结合位点。观察到，FAD的腺苷部分深埋在AOX内，而异恶嗪环则粘附在表面上。本文报道了选择性质子离子载体（CCCP和DNP）与AOX的相互作用，并报道了它们的结合位点。这些质子离子载体显示出与FAD的竞争结合。