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首页> 外文期刊>BioTechnology: An Indian Journal >Keratinolytic activity from new recombinant fusant AYA2000, an endophytic Micromonospora spp.
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Keratinolytic activity from new recombinant fusant AYA2000, an endophytic Micromonospora spp.

机译：来自新型重组融合剂AYA2000（内生微单孢菌属种）的角质蛋白分解活性。

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Two different endophytic strains ESRAA1997 and ALAA2000 were isolated from the Egyptian herbal plant Anastatica hierochuntica. The two strains produced alkaline serine protease and they were identified based on their phenotypic and chemotypic characteristics as two different strains of Micromonospora spp. Both strains grew and produced keratinase using different keratinous waste substances as the sole source of carbon and nitrogen. In our study, the activity and properties of keratinase enzymes of the both wild strains ESRAA1997 and ALAA2000 have been altered by genetic recombination through protoplast fusion between them, leading to a potent keratinolytic fusant Micromonospora spp. AYA2000 with improved properties (activity, stability, specificity and tolerance to inhibitors). Whereas using a mixture of yeast extract, peptone and malt extract as a supplement to the bovine hair medium increased keratinase production by 48%, addition of 1% glucose suppressed enzyme production byMicromonospora spp.AYA2000. The enzyme was purified by ammoniumsulphate precipitation,DEAE-cellulose chromatography followed by gel filtration. The molecular mass, determined using SDS???PAGE, was 39kDa. AYA2000. The enzyme exhibited remarkable activity towards all keratinous wastes used and could also adapt to a broad range of pH and temperature with optima at pH 11 and 60???°C. The enzyme was not influenced by chelating reagents,metal ions or alcohols. These propertiesmake AYA2000 keratinase an ideal candidate for biotechnological application.
机译：从埃及草药植物Anastatica hierochuntica分离出两种不同的内生菌株ESRAA1997和ALAA2000。这两种菌株产生碱性丝氨酸蛋白酶，并且根据它们的表型和化学型特征被鉴定为两种不同的微单孢菌属菌株。两种菌株均使用不同的角质废物作为碳和氮的唯一来源生长并产生角蛋白酶。在我们的研究中，通过野生型ESRAA1997和ALAA2000之间的原生质体融合，通过基因重组改变了野生菌株ESRAA1997和ALAA2000的角蛋白酶的活性和特性，从而导致了强力的角蛋白分解融合剂Micromonospora spp。具有改善的特性（活性，稳定性，特异性和对抑制剂的耐受性）的AYA2000。而使用酵母提取物，蛋白one和麦芽提取物的混合物作为牛毛培养基的补充剂可使角蛋白酶的产生增加48％，而添加1％的葡萄糖则可抑制Micromonospora spp.AYA2000产生的酶。通过硫酸铵沉淀，DEAE-纤维素色谱，然后进行凝胶过滤来纯化酶。用SDS-PAGE测定的分子量为39kDa。 AYA2000。该酶对所用的所有角质废物均显示出显着的活性，并且还可以适应广泛的pH和温度，最适pH为11和60℃。该酶不受螯合剂，金属离子或醇的影响。这些特性使AYA2000角蛋白酶成为生物技术应用的理想候选者。

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