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发明授权
CN107083374B 酶活性提高的β-甘露聚糖酶突变体及其编码基因和应用 有权

酶活性提高的β-甘露聚糖酶突变体及其编码基因和应用
摘要：
本发明提供了酶活性提高的β‑甘露聚糖酶突变体及其编码基因和应用。本发明以能够在巴斯德毕赤酵母Pichia pastoris中进行高效表达的β‑甘露聚糖酶为基础，使用易错PCR方法对该酶基因进行突变，再通过高通量筛选方法挑选出正突变，经过两轮易错PCR，最终获得4个酶活显著提高的突变体，分别为MAN‑M01、MAN‑M02、MAN‑M03和MAN‑M04，其酶活与原始β‑甘露聚糖酶相比，酶活分别提高了40%、58%、80%和64%，本发明获得了酶活性大大提高的β‑甘露聚糖酶，有利于实现其在商业中的应用，从而降低生产成本。
摘要（英）：

The invention provides beta-mannase mutants with improved enzyme activityiesand an encoding gene and application of the beta-mannase mutants. According to the beta-mannase mutants with the improved enzyme activities, based on beta-mannase which can be efficiently expressed in Pichia pastoris, an error-prone PCR method is utilized to mutate an enzyme gene, then through a high throughput screening method, direct mutation is picked out, and through two circles of error-prone PCR, finally four mutants with the remarkably improved enzyme activities are obtained, namely, MAN-M01, MAN-M02, MAN-M03 and MAN-M04 respectively, wherein compared with original beta-mannase, the enzyme activities of the beta-mannase mutants with the improved enzyme activites are increased by 40%, 58%, 80% and 64% respectively. Beta-mannase with the drastically improved enzyme activities obtained, which is beneficial to achieving the application of beta-mannase in commerce, and therefore reduces the production cost.

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