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利用优化的角蛋白酶生产培养基,通过韦氏芽孢杆菌 HFS_F2 对角蛋白废物进行生物降解。

Biodegradation of Keratin Waste by Bacillus velezensis HFS_F2 through Optimized Keratinase Production Medium.

机构信息

Department of Microbial Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, India.

出版信息

Curr Microbiol. 2024 May 18;81(7):179. doi: 10.1007/s00284-024-03699-5.

Abstract

Enormous aggregates of keratinous wastes are produced annually by the poultry and leather industries which cause environmental degradation globally. To combat this issue, microbially synthesized extracellular proteases known as keratinase are used widely which is effective in degrading keratin found in hair and feathers. In the present work, keratinolytic bacteria were isolated from poultry farm soil and feather waste, and various cultural conditions were optimized to provide the highest enzyme production for efficient keratin waste degradation. Based on the primary and secondary screening methods, the potent keratinolytic strain (HFS_F2) with the highest enzyme activity 32.65 ± 0.16 U/mL was genotypically characterized by 16S rRNA sequencing and was confirmed as Bacillus velezensis HFS_F2 ON556508. Through one-variable-at-a-time approach (OVAT), the keratinase production medium was optimized with sucrose (carbon source), beef extract (nitrogen source) pH-7, inoculum size (5%), and incubation at 37 °C). The degree of degradation (%DD) of keratin wastes was evaluated after 35 days of degradation in the optimized keratinase production medium devoid of feather meal under submerged fermentation conditions. Further, the deteriorated keratin wastes were visually examined and the hydrolysed bovine hair with 77.32 ± 0.32% degradation was morphologically analysed through Field Emission Scanning Electron Microscopy (FESEM) to confirm the structural disintegration of the cuticle. Therefore, the current study would be a convincing strategy for reducing the detrimental impact of pollutants from the poultry and leather industries by efficient keratin waste degradation through the production of microbial keratinase.

摘要

每年家禽和制革业都会产生大量的角蛋白废物,这些废物会导致全球环境退化。为了解决这个问题,人们广泛使用微生物合成的细胞外蛋白酶,即角蛋白酶,它可以有效地降解毛发和羽毛中的角蛋白。在本工作中,从家禽养殖场土壤和羽毛废物中分离出角蛋白降解菌,并优化了各种培养条件,以提供最高的酶产量,从而有效地降解角蛋白废物。根据初筛和复筛方法,从具有最高酶活 32.65 ± 0.16 U/mL 的角蛋白降解菌中筛选出高效角蛋白降解菌(HFS_F2),并通过 16S rRNA 测序对其进行了基因型鉴定,鉴定为 Bacillus velezensis HFS_F2 ON556508。通过单因素实验(One-variable-at-a-time approach,OVAT)对产角蛋白酶培养基进行优化,优化后的产酶培养基为蔗糖(碳源)、牛肉浸膏(氮源)pH-7、接种量(5%)和 37℃培养。在优化的角蛋白酶生产培养基中,在 37℃、pH-7 条件下,不添加羽毛粉,进行 35 天的角蛋白降解,评价角蛋白废物的降解程度(%DD)。进一步,在优化的产酶条件下,在静置发酵条件下,对降解 35 天后的变质角蛋白废物进行了外观检查,并对角蛋白水解产物——牛毛进行了形貌分析,发现其角蛋白结构已完全崩解。因此,本研究通过微生物角蛋白酶的生产,为减少家禽和制革业污染物对环境的不利影响提供了一种有效的策略,即高效降解角蛋白废物。

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