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木质分解菌白腐菌 WDP2:漆酶的生产、特性和合成染料的生物修复。

Lignolytic mushroom Lenzites elegans WDP2: Laccase production, characterization, and bioremediation of synthetic dyes.

机构信息

Department of Microbiology, CBSH; G.B. Pant University of Agriculture and Technology Pantnagar, U. S. Nagar, Uttarakhand, India.

Department of Genetics and Plant Breeding, College of Agriculture; G.B. Pant University of Agriculture and Technology Pantnagar, U. S. Nagar, Uttarakhand, India.

出版信息

Ecotoxicol Environ Saf. 2018 Aug 30;158:50-58. doi: 10.1016/j.ecoenv.2018.04.003. Epub 2018 Apr 12.

Abstract

A mycoremedial study was undertaken for decolourization of synthetic dyes using wood rot fungal culture Lenzites elegans WDP2. The culture was isolated from decaying wood as fruiting body, and identified on the basis of 5.8S ITS rRNA gene sequence analysis. Qualitative plate screening of culture showed extracellular laccase and lignin peroxidase production, while only laccase enzyme was produced in higher amount (156.793 Uml) in minimal salt broth medium containing glucose and veratryl alcohol. Laccase activity was increased up to 189.25 Uml after optimization of laccase production by optimization of one variable at a time approach. Molecular characterization of laccase enzyme was done using SDS PAGE and Native PAGE based isozyme analyses. The culture was able to decolorize three synthetic dying compounds (congo red, Malachite green and brilliant green) in broth media, while showed very less decolourization in plate assay. The fungal culture varied in their dye decolourizing potential in broth culture, showing 92.77%, 21.27% and 98.8% maximum decolourization of brilliant green, malachite green and congo red respectively. The congo red dye was completely bio-absorbed by fungal culture within one month. The fungal decolourized broth also revealed the extracellular laccase activity; varied from 10 Uml to 68.5 Uml in all the three cases, supports the involvement of laccase enzyme in decolorization. Phase contrast microscopy clearly revealed bio-sorption of the dyes by fungal culture into the mycelium/spores in the photomicrographs.

摘要

采用木质腐朽真菌栓菌(Lenzites elegans WDP2)培养物对合成染料进行脱色的真菌修复研究。该培养物是从腐烂木材的子实体中分离得到的,并根据 5.8S ITS rRNA 基因序列分析进行了鉴定。定性平板筛选培养物显示胞外漆酶和木质素过氧化物酶的产生,而仅在含有葡萄糖和藜芦醇的低盐肉汤培养基中产生更多的漆酶(156.793 Uml)。通过一次性优化一个变量的方法优化漆酶生产后,漆酶活性提高到 189.25 Uml。使用 SDS-PAGE 和基于 Native-PAGE 的同工酶分析对漆酶酶进行了分子表征。该培养物能够在液体培养基中对三种合成染料化合物(刚果红、孔雀石绿和亮绿)进行脱色,而在平板测定中显示出非常低的脱色率。真菌培养物在液体培养中的染料脱色能力不同,对亮绿、孔雀石绿和刚果红的最大脱色率分别为 92.77%、21.27%和 98.8%。刚果红染料在一个月内被真菌培养物完全生物吸收。真菌脱色的培养液也显示出胞外漆酶活性;在所有三种情况下,从 10 Uml 到 68.5 Uml 不等,这支持了漆酶酶在脱色中的参与。相差显微镜清楚地显示了染料被真菌培养物生物吸附到菌丝/孢子中的情况,在照片中可以看到。

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