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从蚕粪中分离的纤维素降解细菌的特性及其纤维素酶产生的优化

Characterization of Cellulose-Degrading Bacteria Isolated from Silkworm Excrement and Optimization of Its Cellulase Production.

作者信息

Li Hao, Zhang Minqi, Zhang Yuanhao, Xu Xueming, Zhao Ying, Jiang Xueping, Zhang Ran, Gui Zhongzheng

机构信息

College of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.

Sericulture Resources Intensive Processing Laboratory, Sericulture Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212100, China.

出版信息

Polymers (Basel). 2023 Oct 19;15(20):4142. doi: 10.3390/polym15204142.

Abstract

An abundance of refractory cellulose is the key limiting factor restricting the resource utilization efficiency of silkworm () excrement via composting. Screening for cellulose-degrading bacteria is likely to provide high-quality strains for the safe and rapid decomposition of silkworm excrement. In this study, bacteria capable of degrading cellulose with a high efficiency were isolated from silkworm excrement and the conditions for cellulase production were optimized. The strains were preliminarily screened via sodium carboxymethyl cellulose culture and staining with Congo red, rescreened via a filter paper enzyme activity test, and identified via morphological observation, physiological and biochemical tests, and phylogenetic analysis of the 16S rDNA sequence. Enzyme activity assay was performed using the 3,5-dinitrosalicylic acid method. DC-11, a highly cellulolytic strain, was identified as . The optimum temperature and pH of this strain were 55 °C and 6, respectively, and the filter paper enzyme activity (FPase), endoglucanase activity (CMCase), and exoglucanase activity (CXase) reached 15.40 U/mL, 11.91 U/mL, and 20.61 U/mL. In addition, the cellulose degradation rate of the treatment group treated with DC-11 was 39.57% in the bioaugmentation test, which was significantly higher than that of the control group without DC-11 (10.01%). Strain DC-11 was shown to be an acid-resistant and heat-resistant cellulose-degrading strain, with high cellulase activity. This strain can exert a bioaugmentation effect on cellulose degradation and has the potential for use in preparing microbial inocula that can be applied for the safe and rapid composting of silkworm excrement.

摘要

大量难降解纤维素是限制蚕粪堆肥资源利用效率的关键因素。筛选纤维素降解菌可能为蚕粪的安全快速分解提供优质菌株。本研究从蚕粪中分离出高效降解纤维素的细菌,并对纤维素酶的产生条件进行了优化。通过羧甲基纤维素钠培养和刚果红染色进行初步筛选,通过滤纸酶活性测试进行复筛,并通过形态观察、生理生化测试和16S rDNA序列的系统发育分析进行鉴定。采用3,5-二硝基水杨酸法进行酶活性测定。DC-11是一株高纤维素分解菌株,被鉴定为[具体菌种名称未给出]。该菌株的最适温度和pH分别为55℃和6,滤纸酶活性(FPase)、内切葡聚糖酶活性(CMCase)和外切葡聚糖酶活性(CXase)分别达到15.40 U/mL、11.91 U/mL和20.61 U/mL。此外,在生物强化试验中,用DC-11处理的实验组纤维素降解率为39.57%,显著高于未添加DC-11的对照组(10.01%)。菌株DC-11是一株耐酸耐热的纤维素降解菌株,具有较高的纤维素酶活性。该菌株对纤维素降解具有生物强化作用,具有制备可用于蚕粪安全快速堆肥的微生物接种剂的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2295/10610594/3554efd44c23/polymers-15-04142-g001.jpg

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