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枯草芽孢杆菌 SLC 中的角蛋白酶和硫化物回收羽毛废料。

Keratinases and sulfide from Bacillus subtilis SLC to recycle feather waste.

机构信息

Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes (IMPG), Bloco I, Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Cidade Universitária, Ilha do Fundão, Rio de Janeiro, RJ 21941-590, Brazil.

出版信息

World J Microbiol Biotechnol. 2012 Mar;28(3):1259-69. doi: 10.1007/s11274-011-0930-0. Epub 2011 Nov 9.

Abstract

The aim of this study is to investigate the culture conditions of chicken feather degradation and keratinolytic enzyme production by the recently isolated Bacillus subtilis SLC and to evaluate the potential of the SLC strain to recycle feather waste discarded by the poultry industry. The SLC strain was isolated from the agroindustrial waste of a poultry farm in Brazil and was confirmed to belong to Bacillus subtilis by rDNA gene analysis. There was high keratinase production when the medium was at pH 8 (280 U ml(-1)). Activity was higher using the inoculum propagated for 72 h on 1% whole feathers supplemented with 0.1% yeast extract. In the enzymatic extract, the keratinases were active in the pH range from 2.0 to 12.0 with a maximum activity at pH 10.0 and temperature 60°C. For gelatinase the best pH was 5.0 and the best temperature was 37°C. All keratinases are serine peptidases. The crude enzymatic extract degraded keratin, gelatin, casein, and hemoglobin. Scanning electron microscopy showed Bacillus cells adhered onto feather surfaces after 98 h of culture and degraded feather filaments were observed. MALDI-TOF mass spectrometric analysis showed multiple peaks from 522 to 892 m/z indicating feather degradation. The presence of sulfide was detected on extracellular medium probably participating in the breakdown of sulfide bridges of the feather keratin. External addition of sulfide increased feather degradation.

摘要

本研究旨在探讨鸡羽毛降解和角蛋白酶生产的培养条件,由最近分离的枯草芽孢杆菌 SLC 进行,并评估 SLC 菌株回收家禽业丢弃的羽毛废物的潜力。SLC 菌株是从巴西一家家禽养殖场的农业工业废物中分离出来的,通过 rDNA 基因分析证实属于枯草芽孢杆菌。当培养基的 pH 值为 8 时(280 U ml(-1)),角蛋白酶的产量很高。在添加 0.1%酵母提取物的 1%全羽毛上繁殖 72 小时的接种物中,活性更高。在酶提取物中,角蛋白酶在 pH 2.0 至 12.0 范围内具有活性,在 pH 10.0 和温度 60°C 时活性最高。对于明胶酶,最佳 pH 值为 5.0,最佳温度为 37°C。所有角蛋白酶都是丝氨酸肽酶。粗酶提取物可降解角蛋白、明胶、酪蛋白和血红蛋白。扫描电子显微镜显示,培养 98 小时后,芽孢杆菌细胞附着在羽毛表面上,并观察到羽毛纤维降解。MALDI-TOF 质谱分析显示,从 522 到 892 m/z 有多个峰,表明羽毛降解。在细胞外培养基中检测到硫化物的存在,可能参与羽毛角蛋白中硫桥的断裂。外加硫化物可增加羽毛的降解。

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