重组植酸酶在毕赤酵母和多形羊草有利甲基化营养平台上表达的生化特性比较

Ardeshir Hesampour, Omid Ranaei, M. Malboobi, J. Harati, N. Mohandesi
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引用次数: 9

摘要

植酸是植物种子中磷的主要形式。然而,植酸不能被动物利用,导致动物严重缺磷。植酸酶作为饲料添加剂的使用会影响磷光体的释放及其矿物质的利用率。本研究研究了黑孢霉phyA基因在甲基营养酵母中的异源表达。研究了酵母和多形芽孢杆菌及其表达的重组植酸酶的生化和生物物理特性,并与天然黑霉植酸酶进行了比较。设计了黑曲霉的phyA基因序列,合成的基因在两种酵母宿主中均以失活形式成功表达。由于重糖基化,分泌的重组植酸酶的大小从50到65 KDa不等。对表达的细胞外重组植酸酶样品进行了纯化,生化性能测试表明,两种重组植酸酶样品具有相似的pH值,pH最适测定值分别为pH 2.5和pH.5.5,最适温度分别为60°C和50°C。本研究表明,重组巴斯德酵母和多形酵母植酸酶能够满足一系列预先设定的工业质量标准,可作为动物饲料添加剂应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
COMPARISON OF BIOCHEMICAL PROPERTIES OF RECOMBINANT PHYTASE EXPRESSION IN THE FAVORABLE METHYLOTROPHIC PLATFORMS OF PICHIA PASTORIS AND HANSENULA POLYMORPHA
Phytic acid is the dominant form of phosphorous in plant seeds. However, phytic acid cannot beutilized by animals and causes them serious phosphate deficiency. Utilization of phytase as ananimal feed additive can affect liberation of phosphor and its mineral availability. In this study,heterologous expression of the A. niger phyA gene was investigated in the methylotrophic yeastsP. pastoris and H.polymorpha and its expressed active recombinant phytase biochemical andbiophysical properties were studies and compared to native A.niger phytase. The phyA genesequence of A.niger was designed and expression of synthetic genes was highly successfully inactive form in both yeast hosts. Size of the secreted recombinant phytases, due to heavyglycosylation, varied from 50 to 65 KDa. Expressed extracellular recombinant phytase sampleswere purified and biochemical tests on properties demonstrated that both recombinant phytasesamples had similar pH profiles with pH optima determinations of pH 2.5, pH.5.5 as acidphosphatase and optimum temperature of 60 and 50 °C respectively in P. pastoris and H.polymorpha. This study concludes that recombinant P. pastoris and H. polymorpha phytases could fulfil aseries of predefined industrial quality criteria for application as animal feed supplement.
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