巨芽孢杆菌UСM B-5710角化酶的底物特异性

Q4 Biochemistry, Genetics and Molecular Biology Mikrobiolohichnyi zhurnal Pub Date : 2023-10-23 DOI:10.15407/microbiolj85.05.003
K.V. Avdiyuk, L.D. Varbanets
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The culture was grown under conditions of submerged cultivation at 40 °C, with a nutrient medium stirring rate of 201 rpm for 6 days. For growth, a basic nutrient medium containing 0.5% defatted chicken feathers or other keratin-containing substrates as sole sources of carbon and nitrogen were used. Keratinase activity was assessed by UV absorption at 280 nm of hydrolysis products of keratin-containing raw materials. Protein was determined by the Lowry method, caseinolytic (total proteolytic) activity was determined by the Anson method modified by Petrova, and amino acid content was determined by the ninhydrin method. The degree of hydrolysis of the substrates was evaluated by the ratio of the initial and final weight of the substrate. Results. It was shown that the synthesis of keratinase by the culture of B. megaterium UCM B-5710 begins from the 6th hour of cultivation. The level of protein and proteolytic activity and the content of amino acids increased throughout the entire period of culture growth. The supernatant of the culture liquid of B. megaterium UCM B-5710 was most effective in splitting white chicken’s and turkey’s feathers, a little slower — feathers of black chicken and blue parrots, as well as wool of white sheep. According to the degree of splitting, the substrates used can be arranged in the following order: white turkey feathers > white chicken feathers > black chicken feathers > blue parrot feathers > white sheep wool > baby nails > pig bristle > baby hair. The study of the effect of feather color on the resistance to decomposition showed that black, blue, and red feathers are more resistant, which coincides with the literature data. Conclusions. 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引用次数: 0

摘要

畜禽产品加工的具体情况是,在获得主要适销产品的过程中,在工艺过程的各个阶段,大约有一半的原料变成污染环境的废物。这些副产品含有大量难以消化的角蛋白。使用能够降解这种蛋白质的特定酶不仅有助于减少对自然的负面人为影响,而且还有助于获得可用作植物肥料或饲料添加剂的有价值的水解物。本研究的目的是研究巨芽孢杆菌UCM B-5710对各种含角蛋白底物的裂解能力:黑白鸡毛、白火鸡羽毛、各种颜色的鹦鹉羽毛、羊毛、猪鬃毛、婴儿毛发和指甲。方法。培养物在40℃深层培养条件下,以201转/分的营养培养基搅拌速度培养6天。为了生长,使用含有0.5%脱脂鸡毛或其他含角蛋白的基质作为碳和氮的唯一来源的基本营养培养基。用含角蛋白原料的水解产物在280 nm处紫外吸收评价角蛋白酶活性。蛋白质用Lowry法测定,酪蛋白水解(总蛋白水解)活性用Petrova改良的Anson法测定,氨基酸含量用茚三酮法测定。底物的水解程度通过底物的初始重量和最终重量的比值来评价。结果。结果表明,大芽孢杆菌UCM B-5710从培养第6小时开始合成角化酶。蛋白质水平、蛋白水解活性和氨基酸含量在整个培养生长期间均呈上升趋势。megaterium UCM B-5710培养液上清液对白鸡和火鸡羽毛的分裂效果最好,对黑鸡和蓝鹦鹉羽毛的分裂效果稍慢,对白羊羊毛的分裂效果较差。根据劈裂程度,所使用的基材可按以下顺序排列:白火鸡羽毛>白色的鸡毛>黑鸡毛>蓝色鹦鹉羽毛>白羊毛>婴儿指甲>猪鬃;婴儿的头发。羽毛颜色对抗分解能力影响的研究表明,黑色、蓝色和红色羽毛的抗分解能力更强,这与文献数据相吻合。结论。然而,B. megaterium UCM B-5710产生的角化酶能够以不同的效率和速率分裂α-和β-角化蛋白。
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Substrate Specificity of Bacillus megaterium UСM B-5710 Keratinase
The specifics of the processing of livestock and poultry products is that in the process of obtaining the main marketable products, about half the feedstock at various stages of the technological process turns into waste that pollutes the environment. These by-products contain large amounts of the hard-to-digest keratin protein. The use of specific enzymes capable of degrading this protein helps not only to reduce the negative anthropogenic impact on nature but also to obtain valuable hydrolysates that can be used as a fertilizer for plants or a feed additive. The aim of this work was to study the ability of Bacillus megaterium UCM B-5710 to split various keratin-containing substrates: black and white chicken feathers, white turkey feathers, parrot feathers of various colors, sheep wool, pig bristles, and baby hair and nails. Methods. The culture was grown under conditions of submerged cultivation at 40 °C, with a nutrient medium stirring rate of 201 rpm for 6 days. For growth, a basic nutrient medium containing 0.5% defatted chicken feathers or other keratin-containing substrates as sole sources of carbon and nitrogen were used. Keratinase activity was assessed by UV absorption at 280 nm of hydrolysis products of keratin-containing raw materials. Protein was determined by the Lowry method, caseinolytic (total proteolytic) activity was determined by the Anson method modified by Petrova, and amino acid content was determined by the ninhydrin method. The degree of hydrolysis of the substrates was evaluated by the ratio of the initial and final weight of the substrate. Results. It was shown that the synthesis of keratinase by the culture of B. megaterium UCM B-5710 begins from the 6th hour of cultivation. The level of protein and proteolytic activity and the content of amino acids increased throughout the entire period of culture growth. The supernatant of the culture liquid of B. megaterium UCM B-5710 was most effective in splitting white chicken’s and turkey’s feathers, a little slower — feathers of black chicken and blue parrots, as well as wool of white sheep. According to the degree of splitting, the substrates used can be arranged in the following order: white turkey feathers > white chicken feathers > black chicken feathers > blue parrot feathers > white sheep wool > baby nails > pig bristle > baby hair. The study of the effect of feather color on the resistance to decomposition showed that black, blue, and red feathers are more resistant, which coincides with the literature data. Conclusions. B. megaterium UCM B-5710 produces keratinase capable of splitting both α- and β-keratins, however, with different efficiencies and rates.
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Mikrobiolohichnyi zhurnal
Mikrobiolohichnyi zhurnal Medicine-Microbiology (medical)
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