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研究在浸没培养中生产碱性角蛋白酶作为皮革技术中产生的固体废物处理的替代方法。

Study of the production of alkaline keratinases in submerged cultures as an alternative for solid waste treatment generated in leather technology.

机构信息

Research and Development Center for Industrial Fermentations, Calle 47 y 115 (B1900ASH), La Plata, Argentina.

出版信息

J Microbiol Biotechnol. 2013;23(7):1004-14. doi: 10.4014/jmb.1211.11016.

Abstract

Six nonpathogenic fungal strains isolated from alkaline soils of Buenos Aires Province, Argentina (Acremonium murorum, Aspergillus sidowii, Cladosporium cladosporoides, Neurospora tetrasperma, Purpureocillium lilacinum (formerly Paecilomyces lilacinus), and Westerdikella dispersa) were tested for their ability to produce keratinolytic enzymes. Strains were grown on feather meal agar as well as in solid-state and submerged cultures, using a basal mineral medium and "hair waste" as sole sources of carbon and nitrogen. All the tested fungi grew on feather meal agar, but only three of them were capable of hydrolyzing keratin, producing clear zones. Among these strains, P. lilacinum produced the highest proteolytic and keratinolytic activities, both in solid-state and submerged fermentations. The medium composition and culture conditions for the keratinases production by P. lilacinum were optimized. Addition of glucose (5 g/l) and yeast extract (2.23 g/l) to the basal hair medium increased keratinases production. The optimum temperature and initial pH for the enzyme production were 28℃ and 6.0, respectively. A beneficial effect was observed when the original concentration of four metal ions, present in the basal mineral medium, was reduced up to 1:10. The maximum yield of the enzyme was 15.96 Uc/ml in the optimal hair medium; this value was about 6.5-fold higher than the yield in the basal hair medium. These results suggest that keratinases from P. lilacinum can be useful for biotechnological purposes such as biodegradation (or bioconversion) of hair waste, leading to a reduction of the environmental pollution caused by leather technology with the concomitant production of proteolytic enzymes and protein hydrolyzates.

摘要

从阿根廷布宜诺斯艾利斯省碱性土壤中分离出的 6 株非致病真菌菌株(枝顶孢属、Sidowii 曲霉、枝孢属、四孢神经霉、淡紫拟青霉(原拟青霉属)和分散西地霉)被测试其产生角蛋白水解酶的能力。菌株在羽毛粉琼脂以及固态和液态培养中生长,使用基础矿物培养基和“毛发废物”作为唯一的碳源和氮源。所有测试的真菌都在羽毛粉琼脂上生长,但只有其中 3 株能够水解角蛋白,产生透明区。在这些菌株中,淡紫拟青霉在固态和液态发酵中均产生最高的蛋白水解酶和角蛋白水解酶活性。优化了淡紫拟青霉角蛋白酶生产的培养基组成和培养条件。在基础毛发培养基中添加葡萄糖(5 g/l)和酵母提取物(2.23 g/l)可提高角蛋白酶的产量。酶生产的最佳温度和初始 pH 值分别为 28℃和 6.0。当基础矿物培养基中存在的四种金属离子的原始浓度降低至 1:10 时,观察到有益的效果。在最佳毛发培养基中,酶的最大产量为 15.96 Uc/ml;这一数值比在基础毛发培养基中的产量高约 6.5 倍。这些结果表明,淡紫拟青霉的角蛋白酶可用于生物技术目的,如毛发废物的生物降解(或生物转化),从而减少皮革技术造成的环境污染,并伴随着蛋白水解酶和蛋白质水解产物的产生。

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