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高温真菌嗜热毁丝霉木聚糖酶基因在巴斯德毕赤酵母中的高效表达。

High-level expression of a xylanase gene from the thermophilic fungus Paecilomyces thermophila in Pichia pastoris.

机构信息

Department of Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Qinghua Donglu, Haidian District, Beijing, China.

出版信息

Biotechnol Lett. 2012 Nov;34(11):2043-8. doi: 10.1007/s10529-012-0995-3. Epub 2012 Jul 11.

Abstract

A xylanase gene from Paecilomyces thermophila was functionally expressed in Pichia pastoris. The recombinant xylanase (xynA) was predominantly extracellular; in a 5 l fermentor culture, the total extracellular protein was 8.1 g l(-1) with an activity of 52,940 U ml(-1). The enzyme was purified to homogeneity with a recovery of 48 %. The recombinant xynA was optimally active at 75 °C, as measured over 10 min, and at pH 7. The enzyme was stable up to 80 °C for 30 min. It hydrolyzed birchwood xylan, beechwood xylan and xylooligosaccharides to produce xylobiose and xylotriose as the main products.

摘要

嗜热毁丝霉木聚糖酶基因在巴斯德毕赤酵母中实现了功能表达。重组木聚糖酶(xynA)主要为胞外酶;在 5L 发酵罐培养中,总胞外蛋白为 8.1g/L,酶活为 52940U/ml。该酶经纯化达到均一性,回收率为 48%。重组 xynA 的最适活性温度为 75°C,在 10 分钟内测量,最适 pH 值为 7。该酶在 80°C 下稳定 30 分钟。它可以水解桦木木聚糖、山毛榉木聚糖和木寡糖,生成主要产物木二糖和木三糖。

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