使用基于PNP的试剂选新型葡萄糖乙酶的高效活性选
Michael S Madsen1, Pedro A Martins1, Jane W Agger1
1Technical University of Denmark, Lignin Biotechnology, Department of Biotechnology and Biomedicine, Søltofts Plads 224, Kgs Lyngby DK-2800, Denmark.
Enzyme and microbial technology
|April 6, 2024
概括
研究人员选了15种真菌葡萄糖乙酶 (CE15) 进行菌纤维素加工. 七种酶表现出活性,其中活性候选物来自Auricularia subglabra,Ganoderma sinensis和Neocallimastix californiae.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 生物技术是生物技术.
背景情况:
- 葡萄糖乙酶 (CE15) 对于分解纤维素酸至关重要.
- 识别新型CE15酶对于高效的纤维素蛋白加工至关重要.
- 目前的选方法受到基质表现不佳的限制.
研究的目的:
- 为了表达和选15种新的真菌CE15蛋白质以检测雌激酶活性.
- 开发和使用一种改进的色度测定试验,用于酶查.
- 为潜在的工业应用确定活跃的CE15候选者.
主要方法:
- 15种真菌CE15蛋白的表达和净化.
- 使用甲基-UX-β-pNP基质进行色度选.
- 活动测定与GH67和GH43酶相结合.
- 确定活性酶的特异性活性.
主要成果:
- 在15个表达的CE15蛋白质中,有7个表现出可检测的活性.
- 在 Auricularia subglabra (3), Ganoderma sinensis (2) 和 Neocallimastix californiae (2) 中发现了活性酶.
- 来自S. commune,P. anserina,T. versicolor和C. cinerea的CE15蛋白在测试基质上是不活性的.
结论:
- 一种新的色度测定方法有效地选葡萄糖烯酸乙酶活性.
- 鉴定了几种活跃的真菌CE15酶,为林氏纤维素增值提供了潜在的潜力.
- 一些已知的纤维蛋白降解剂中意想不到的缺乏活性,需要进一步调查基质特异性.
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