Bohacz Justyna
Department of Environmental Microbiology, Laboratory of Mycology, Faculty of Agrobioengineering, University of Life Sciences in Lublin, 7 Leszczyńskiego Street, 20-069, Lublin, Poland.
World J Microbiol Biotechnol. 2017 Jan;33(1):13. doi: 10.1007/s11274-016-2177-2. Epub 2016 Nov 24.
This paper assesses the ability of strains of Aphanoascus fulvescens and Chrysosporium articulatum isolated from soil (phaesol) to degrade native feather keratin. Strains were identified based on phenotypic traits and nucleotide sequencing. Response Surface Methodology was used to optimize cultivation conditions exhibiting the highest keratinolytic activity. The experiments were based on Box-Behnken designs for the loss of substrate mass (chicken feathers). While substrate mass loss is an "economic coefficient" that reliably indicates feather keratin degradation, it has not been studied before. Stationary liquid cultures of five selected strains were conducted in laboratory conditions at 28 °C using poultry feathers (1 g) as the sole source of carbon, nitrogen and energy. Enzymatic activities, keratin mineralization products and substrate mass loss were determined periodically. The mineralization of keratin proteins by strains yielded a high number of ammonium ions alkalinizing the medium. Increased ammonium ions inhibited the activity of caseinian protease and keratinase. A decrease in the concentration of these ions induced proteolytic enzymes, chiefly the activity of keratinase, at the end of fungal cultivation. Keratinase activity was related to protein- and peptide release and that of caseinian protease to sulfate ions. The highest loss of substrate mass in comparison to the reference strain CBS104.62 (35.4%) was recorded for Aphanoascus fulvescens B21/4-5 (65.9%). Based on a Box-Behnken design, the maximum loss of substrate mass for the Aphanoascus fulvescens strain (71.08%) can be achieved at pH 7.58 and temperature 28.7 °C.
本文评估了从土壤(菜豆)中分离出的黄曲霉和节状金孢子菌菌株降解天然羽毛角蛋白的能力。菌株基于表型特征和核苷酸测序进行鉴定。采用响应面法优化表现出最高角蛋白分解活性的培养条件。实验基于Box-Behnken设计,以底物质量(鸡毛)的损失为指标。虽然底物质量损失是可靠指示羽毛角蛋白降解的“经济系数”,但此前尚未对其进行研究。在实验室条件下,于28℃使用家禽羽毛(1克)作为唯一碳、氮和能量来源,对五个选定菌株进行静置液体培养。定期测定酶活性、角蛋白矿化产物和底物质量损失。菌株对角蛋白的矿化产生了大量使培养基碱化的铵离子。铵离子增加会抑制酪蛋白蛋白酶和角蛋白酶的活性。在真菌培养结束时,这些离子浓度降低会诱导蛋白水解酶,主要是角蛋白酶的活性。角蛋白酶活性与蛋白质和肽的释放有关,酪蛋白蛋白酶活性与硫酸根离子有关。与参考菌株CBS104.62相比,黄曲霉B21/4-5的底物质量损失最高(65.9%),而参考菌株的底物质量损失为35.4%。基于Box-Behnken设计,在pH 7.58和温度28.7℃时,黄曲霉菌株的底物质量损失可达到最大值(71.08%)。