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新型巴氏芽孢杆菌 AR57 产热耐溶剂碱性丝氨酸角蛋白酶的可持续生产、生化和分子特性及其在有前景的家禽固体废弃物管理中的应用。

Sustainable production, biochemical and molecular characterization of thermo-and-solvent stable alkaline serine keratinase from novel Bacillus pumilus AR57 for promising poultry solid waste management.

机构信息

Department of Biotechnology, University College of Engineering, Anna University - BIT Campus, Tiruchirappalli, Tamilnadu 620 024, India.

Department of Biotechnology, University College of Engineering, Anna University - BIT Campus, Tiruchirappalli, Tamilnadu 620 024, India.

出版信息

Int J Biol Macromol. 2020 Nov 15;163:135-146. doi: 10.1016/j.ijbiomac.2020.06.219. Epub 2020 Jun 29.

Abstract

The increasing amount of recalcitrant keratinous wastes generated from the poultry industry poses a serious threat to the environment. Keratinase have gained much attention to convert these wastes into valuable products. Ever since primitive feathers first appeared on dinosaurs, microorganisms have evolved to degrade this most recalcitrant keratin. In this study, we identified a promising keratinolytic bacterial strain for bioconversion of poultry solid wastes. A true keratinolytic bacterium was isolated from the slaughterhouse soil and was identified and designated as Bacillus pumilus AR57 by 16S rRNA sequencing. For enhanced keratinase production and rapid keratin degradation, the media components and substrate concentration were optimized through shake flask culture. White chicken feather (1% w/v) was found to be the good substrate concentration for high keratinase production when supplemented with simple medium ingredients. The biochemical characterization reveals astounding results which makes the B. pumilus AR57 keratinase as a novel and unique protease. Optimum activity of the crude enzyme was exhibited at pH 9 and 45 °C. The crude extracellular keratinase was characterized as thermo-and-solvent (DMSO) stable serine keratinase. Bacillus pumilus AR57 showed complete degradation (100%) of white chicken feather (1% w/v) within 18 h when incubated in modified minimal medium supplemented with DMSO (1% v/v) at 150 rpm at 37 °C. Keratinase from modified minimal medium supplemented with DMSO exhibits a half-life of 4 days. Whereas, keratinase from the modified minimal medium fortified with white chicken feather (1% w/v) was stable for 3 h only. Feather meal produced by B. pumilus AR57 was found to be rich in essential amino acids. Hence, we proposed B. pumilus AR57 as a potential candidate for the future application in eco-friendly bioconversion of poultry waste and the keratinase could play a pivotal role in the detergent industry. While feather meal may serve as an alternative to produce animal feed and biofertilizers.

摘要

从家禽业产生的大量难以处理的角蛋白废物对环境构成了严重威胁。角蛋白酶已引起广泛关注,可将这些废物转化为有价值的产品。自从恐龙最初出现原始羽毛以来,微生物就已经进化到可以降解这种最顽固的角蛋白。在这项研究中,我们鉴定了一种有前途的用于生物转化家禽固体废物的角蛋白降解细菌菌株。从屠宰场土壤中分离出一种真正的角蛋白降解细菌,并通过 16S rRNA 测序鉴定并命名为解淀粉芽孢杆菌 AR57。为了提高角蛋白酶的产量和快速降解角蛋白,通过摇瓶培养优化了培养基成分和底物浓度。发现当补充简单的培养基成分时,白鸡羽毛(1%w/v)是高角蛋白酶产量的良好底物浓度。生化特性分析得出了惊人的结果,使解淀粉芽孢杆菌 AR57 角蛋白酶成为一种新型独特的蛋白酶。粗酶的最适活性在 pH 9 和 45°C 下表现出。粗细胞外角蛋白酶的特性为耐热和耐溶剂(DMSO)稳定的丝氨酸角蛋白酶。在 37°C 下,在补充有 DMSO(1%v/v)的改良最小培养基中以 150rpm 孵育时,Bacillus pumilus AR57 在 18 小时内可完全降解(100%)白鸡羽毛(1%w/v)。添加 DMSO 的改良最小培养基中的角蛋白酶半衰期为 4 天。而在改良最小培养基中添加白鸡羽毛(1%w/v)的角蛋白酶仅稳定 3 小时。发现由解淀粉芽孢杆菌 AR57 产生的羽毛粉富含必需氨基酸。因此,我们提出解淀粉芽孢杆菌 AR57 是未来在环保生物转化家禽废物方面的潜在候选者,角蛋白酶在洗涤剂行业中可能发挥关键作用。而羽毛粉可能可以作为生产动物饲料和生物肥料的替代品。

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