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从森林土壤中分离出一株多功能羽毛降解枯草芽孢杆菌的特性研究。

Characterization of a multifunctional feather-degrading Bacillus subtilis isolated from forest soil.

机构信息

Department of Life Science and Environmental Biochemistry, Pusan National University, Republic of Korea.

出版信息

Biodegradation. 2010 Nov;21(6):1029-40. doi: 10.1007/s10532-010-9363-y. Epub 2010 May 8.

Abstract

In this study, we isolated and characterized a novel feather-degrading bacterium that shows keratinolytic, antifungal and plant growth-promoting activities. A bacterium S8 was isolated from forest soil and confirmed to belong to Bacillus subtilis by BIOLOG system and 16S rRNA gene analysis. The improved culture conditions for the production of keratinolytic protease were 0.1% (w/v) sorbitol, 0.3% (w/v) KNO(3), 0.1% (w/v) K(2)HPO(4), 0.06% (w/v) KH(2)PO(4) and 0.04% (w/v) MgCl(2)·6H(2)O (pH 8.0 and 30°C), respectively. In the improved medium containing 0.1% (w/v) feather, keratinolytic protease production was around 53.3 ± 0.3 U/ml at 4 day; this value was 10-fold higher than the yield in the basal feather medium (5.3 ± 0.1 U/ml). After cultivation for 5 days in the improved medium, intact feather was completely degraded. Feather degradation resulted in free -SH group, soluble protein and amino acids production. The concentration of free -SH group in the culture medium was 15.5 ± 0.2 μM at 4 days. Nineteen amino acids including all essential amino acids were produced in the culture medium; the concentration of total amino acid produced was 3360.4 μM. Proline (2809.9 μM), histidine (371.3 μM) and phenylalanine (172.0 μM) were the major amino acids released in the culture medium. B. subtilis S8 showed the properties related to plant growth promotion: hydrolytic enzymes, ammonification, indoleacetic acid (IAA), phosphate solubilization, and broad-spectrum antimicrobial activity. Interestingly, the strain S8 grown in the improved medium produced IAA and antifungal activity, indicating simultaneous production of keratinolytic and antifungal activities and IAA by B. subtilis S8. These results suggest that B. subtilis S8 could be not only used to improve the nutritional value of feather wastes but also is useful in situ biodegradation of feather wastes. Furthermore, it could also be a potential biofertilizer or biocontrol agent applicable to crop plant soil.

摘要

在这项研究中,我们分离并鉴定了一种新型的羽毛降解菌,该菌具有角蛋白水解、抗真菌和促进植物生长的活性。一种名为 S8 的细菌从森林土壤中分离出来,通过 BIOLOG 系统和 16S rRNA 基因分析证实属于枯草芽孢杆菌。角蛋白水解酶产生的最佳培养条件为:0.1%(w/v)山梨醇、0.3%(w/v)硝酸钾、0.1%(w/v)磷酸氢二钾、0.06%(w/v)磷酸二氢钾和 0.04%(w/v)氯化镁六水合物(pH8.0 和 30°C)。在含有 0.1%(w/v)羽毛的改良培养基中,4 天时角蛋白水解酶的产量约为 53.3±0.3 U/ml;这一数值比基础羽毛培养基中的产量(5.3±0.1 U/ml)高 10 倍。在改良培养基中培养 5 天后,完整的羽毛完全降解。羽毛降解导致 -SH 基团、可溶性蛋白和氨基酸的产生。4 天时培养基中游离 -SH 基团的浓度为 15.5±0.2 μM。在培养基中产生了 19 种氨基酸,包括所有必需氨基酸;总氨基酸的浓度为 3360.4 μM。脯氨酸(2809.9 μM)、组氨酸(371.3 μM)和苯丙氨酸(172.0 μM)是培养基中释放的主要氨基酸。枯草芽孢杆菌 S8 具有与植物生长促进相关的特性:水解酶、氨化、吲哚乙酸(IAA)、磷酸盐溶解和广谱抗菌活性。有趣的是,在改良培养基中生长的菌株 S8 产生了 IAA 和抗真菌活性,表明枯草芽孢杆菌 S8 同时产生角蛋白水解和抗真菌活性以及 IAA。这些结果表明,枯草芽孢杆菌 S8 不仅可用于提高羽毛废物的营养价值,而且还可原位生物降解羽毛废物。此外,它还可以作为一种有潜力的生物肥料或生物防治剂,适用于作物植物土壤。

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