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-Nitrophenol 烷酸酯基测定法在生物塑料降解用微生物菌株的土壤选择和筛选中的双重应用。

Dual Application of -Nitrophenol Alkanoate-Based Assay for Soil Selection and Screening of Microbial Strains for Bioplastic Degradation.

机构信息

Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea.

Institute for Ubiquitous Information Technology and Application, Konkuk University, Seoul 05029, Republic of Korea.

出版信息

J Microbiol Biotechnol. 2024 Jul 28;34(7):1530-1543. doi: 10.4014/jmb.2403.03013. Epub 2024 May 30.

Abstract

With an increase in the commercialization of bioplastics, the importance of screening for plastic-degrading strains and microbes has emerged. Conventional methods for screening such strains are time-consuming and labor-intensive. Therefore, we suggest a method for quickly and effectively screening plastic-degrading microbial strains through dual esterase assays for soil and isolated strains, using -nitrophenyl alkanoates as substrates. To select microbe-abundant soil, the total amount of phospholipid fatty acids (PLFAs) included in each soil sample was analyzed, and esterase assays were performed for each soil sample to compare the esterase activity of each soil. In addition, by analyzing the correlation coefficients and sensitivity between the amount of PLFAs and the degree of esterase activity according to the substrate, it was confirmed that substrate NP-C2 is the most useful index for soil containing several microbes having esterase activity. In addition, esterase assays of the isolated strains allowed us to select the most active strain as the degrading strain, and 16S rRNA results confirmed that it was sp. N04 showed the highest degradation activity for polybutylene succinate (PBS) as measured in liquid culture for 7 days, with a degradation yield of 99%. Furthermore, sp. N04 showed degradation activity against various bioplastics. We propose the dual application of -nitrophenyl alkanoates as an efficient method to first select the appropriate soil and then to screen for plastic-degrading strains in it, and conclude that NP-C2 in particular, is a useful indicator.

摘要

随着生物塑料的商业化程度不断提高,筛选能够降解塑料的菌株和微生物变得越来越重要。传统的筛选这些菌株的方法既耗时又费力。因此,我们建议通过使用 - 硝基苯烷酸酯作为底物,对土壤和分离菌株进行双重酯酶分析,从而快速有效地筛选出能够降解塑料的微生物菌株。为了选择微生物丰富的土壤,分析了每个土壤样本中总磷脂脂肪酸(PLFA)的含量,并对每个土壤样本进行酯酶分析,以比较每个土壤样本的酯酶活性。此外,根据底物分析了 PLFA 含量与酯酶活性之间的相关系数和敏感性,证实了 NP-C2 是含有多种具有酯酶活性的微生物的土壤最有用的指标。此外,对分离菌株的酯酶分析可以选择最活跃的菌株作为降解菌株,16S rRNA 结果证实,在液体培养中, sp. N04 对聚丁二酸丁二醇酯(PBS)的降解活性最高,在 7 天内的降解率达到 99%。此外, sp. N04 对各种生物塑料都表现出降解活性。我们提出将 - 硝基苯烷酸酯双重应用作为一种有效的方法,首先选择合适的土壤,然后在其中筛选能够降解塑料的菌株,并得出结论,特别是 NP-C2,是一种有用的指标。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78c8/11294652/cc136810b664/jmb-34-7-1530-f1.jpg

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