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Identification and characterisation of a Bacillus licheniformis strain with profound keratinase activity for degradation of melanised feather

机译:鉴定和鉴定具有深远角蛋白酶活性的地衣芽孢杆菌菌株以降解黑色化的羽毛

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摘要

Significant amount of keratins in the form of feather, hair, hoof and horn are generated annually by the livestock industry. Keratinases are increasingly important in the reprocessing and environmental pollution control of keratin wastes. The aim of this study is to isolate a microbial strain of high keratinase activity and to evaluate its feather degrading potential. Thirty-two keratin degrading microbial strains from farmyard wastes and primary effluent were isolated using a selective medium containing feather meal at 30, 37 and 50 °C. One of the isolates, which demonstrated the highest keratinolytic activity (11.00 ± 0.71 Uml-1) was identified as a species of Bacillus licheniformis based on the 16S rDNA analysis, designated as strain N22 and deposited in a culture collection. Optimum keratinase production by this bacterium was achieved in 32 h using a minimum growth medium containing 1.1% (w/v) feather meal at 50 °C and pH 8.5. The molecular weight of the keratinase was ≈ 28 KDa as determined sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The keratinase reported here significantly degraded melanised feather in 48 h in the absence of reducing agents. There are few reports on the evaluation of feather degrading ability of keratinases using highly resistant melanised feather. The efficient degradation of melanised feathers by this keratinase may offer an environmentally friendly solution to the degradation of feather waste and other organic matter of similar molecular composition.
机译:畜牧业每年产生大量以羽毛,头发,蹄和角形式的角蛋白。角蛋白酶在角蛋白废物的再加工和环境污染控制中越来越重要。这项研究的目的是分离具有高角蛋白酶活性的微生物菌株,并评估其羽毛降解潜力。使用含有羽毛粉的选择性培养基在30、37和50°C的温度下,从农家废物和主要废水中分离出32种降解角蛋白的微生物菌株。根据16S rDNA分析,分离出的具有最高角蛋白分解活性(11.00±0.71 Uml-1)的一种菌株被鉴定为地衣芽孢杆菌,命名为N22菌株,并保存在培养物中。在50°C和pH值为8.5的条件下,使用含有1.1%(w / v)羽毛粉的最小生长培养基,可在32小时内实现该细菌的最佳角蛋白酶生产。经测定的十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析并通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)证实,角蛋白酶的分子量约为28 KDa。在没有还原剂的情况下,此处报道的角蛋白酶在48小时内可显着降解黑色素化羽毛。关于使用高度抗性的黑色化羽毛来评价角蛋白酶的羽毛降解能力的报道很少。该角蛋白酶有效降解黑化的羽毛可为降解羽毛废料和类似分子组成的其他有机物质提供环境友好的解决方案。

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