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新型菌株sp. DAB-1Y和sp. DAB-1W对林丹的降解研究

Degradation study of lindane by novel strains sp. DAB-1Y and sp. DAB-1W.

作者信息

Kumar Dharmender, Kumar Abhijit, Sharma Jyoti

机构信息

Department of Biotechnology, Deenbandhu Chhotu Ram University of Science and Technology, Murthal, Sonepat, Haryana 131039 India.

出版信息

Bioresour Bioprocess. 2016;3(1):53. doi: 10.1186/s40643-016-0130-8. Epub 2016 Dec 28.

Abstract

BACKGROUND

This study was carried out to isolate and characterize the bacterial strains from lindane-contaminated soil and they were also assessed for their lindane-degrading potential.

METHODS

In this study the enrichment culture method was used for isolation of  lindane degrading bacterial isolates, in which the mineral salt medium (MSM) supplemented with different concentrations of lindane was used. Further, the screening for the potential lindane degrading isolates was done using the spray plate method and colorimetric dechlorinase enzyme assay. The selected isolates were also studied for their growth response under varying range of temperature, pH, and NaCl. The finally selected isolates DAB-1Y and DAB-1W showing best lindane degradation activity was further subjected to biochemical characterization, microscopy, degradation/kinetic study, and 16S rDNA sequencing. The strain identification were performed using the biochemical characterization, microscopy and the species identifies by 16S rDNA sequence of the two isolates using the standard 16S primers, the 16 S rRNA partial sequence was analyzed through BLAST analysis and phylogenetic tree was generated based on UGPMA clustering method using MEGA7 software. This shows the phylogenetic relationship with the related strains. The two isolates of this study were finally characterized as sp. DAB-1Y and sp. DAB-1W, and their 16S rRNA sequence was submitted to GenBank database with accession numbers, KJ811539 and KX986577, respectively.

RESULTS

Out of the 20 isolates, the isolates DAB-1Y and DAB-1W exhibited best lindane-degrading activity of 94 and 98%, respectively, recorded after 8 days of incubation. The optimum growth was observed at temperature 30 °C, pH 7, and 5% NaCl observed for both isolates. Of the four isomers of hexachlorocyclohexane, isomer α and γ were the fastest degrading isomers, which were degraded up to 86 and 94% by isolates DAB-1Y and up to 93 and 98% by DAB-1W, respectively, reported after 8 days incubation. Isomer β was highly recalcitrant in which maximum 35 and 32% lindane degradation was observed even after 28 days incubation by isolates, DAB-1Y and DAB-1W, respectively. At lower lindane concentrations (1-10 mg/L), specific growth rate increased with increase in lindane concentration, maximum being 0.008 and 0.006/day for DAB-1Y and DAB-1W, respectively. The 16 S rRNA partial sequence of isolate DAB-1Y showed similarity with sp. by BLAST analysis and was named as sp. DAB-1Y and DAB-IW with sp. DAB-1W. The 16S rDNA sequence of isolate DAB-1Y and DAB-1W was submitted to online at National Centre of Biotechnology Information (NCBI) with GenBank accession numbers, KJ811539 and KX986577, respectively.

CONCLUSIONS

This study has demonstrated that sp. DAB-1Y and sp. DAB-1W were found efficient in bioremediation of gamma-HCH and can be utilized further for biodegradation of environmental contamination of lindane and can be utilized in bioremediation program.

摘要

背景

本研究旨在从林丹污染土壤中分离和鉴定细菌菌株,并评估它们的林丹降解潜力。

方法

本研究采用富集培养法分离林丹降解细菌菌株,使用添加不同浓度林丹的矿物盐培养基(MSM)。此外,采用喷雾平板法和比色脱氯酶测定法筛选潜在的林丹降解菌株。还研究了所选菌株在不同温度、pH值和NaCl浓度范围内的生长反应。最终筛选出的具有最佳林丹降解活性的菌株DAB-1Y和DAB-1W,进一步进行生化特性分析、显微镜观察、降解/动力学研究以及16S rDNA测序。使用生化特性分析、显微镜观察进行菌株鉴定,并使用标准16S引物通过两个分离株的16S rDNA序列鉴定物种,通过BLAST分析对16S rRNA部分序列进行分析,并使用MEGA7软件基于UGPMA聚类方法生成系统发育树。这显示了与相关菌株的系统发育关系。本研究的两个分离株最终被鉴定为DAB-1Y菌属和DAB-1W菌属,其16S rRNA序列分别提交至GenBank数据库,登录号分别为KJ811539和KX986577。

结果

在20个分离株中,菌株DAB-1Y和DAB-1W在培养8天后表现出最佳的林丹降解活性,分别为94%和98%。两个菌株在温度30°C、pH值7和5% NaCl条件下观察到最佳生长。在六氯环己烷的四种异构体中,异构体α和γ是降解最快的异构体,培养8天后,菌株DAB-1Y对其降解率分别高达86%和94%,菌株DAB-1W对其降解率分别高达93%和98%。异构体β具有高度抗性,即使在培养28天后,菌株DAB-1Y和DAB-1W对其的最大林丹降解率分别仅为35%和32%。在较低的林丹浓度(1 - 10 mg/L)下,比生长速率随林丹浓度增加而增加,DAB-1Y和DAB-1W的最大值分别为0.008和0.006/天。通过BLAST分析,菌株DAB-1Y的16S rRNA部分序列与某菌属显示出相似性,并被命名为DAB-1Y菌属和DAB-1W菌属。菌株DAB-1Y和DAB-1W的16S rDNA序列分别在线提交至美国国立生物技术信息中心(NCBI),GenBank登录号分别为KJ811539和KX986577。

结论

本研究表明,DAB-1Y菌属和DAB-1W菌属在林丹的生物修复中具有高效性,可进一步用于林丹环境污染的生物降解,并可应用于生物修复计划。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/153a/5196013/8788fba47f8c/40643_2016_130_Fig1_HTML.jpg

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