优化后的哈茨木霉 PP389612 胞外漆酶的动力学研究及其对偶氮染料去除能力。

Kinetic studies on optimized extracellular laccase from Trichoderma harzianum PP389612 and its capabilities for azo dye removal.

机构信息

Molecular Biology Research and Studies Institute, Assiut University, Assiut, Egypt.

Botany and Microbiology Department, Faculty of Science, Assiut University, P.O. 71516, Assiut, Egypt.

出版信息

Microb Cell Fact. 2024 May 24;23(1):150. doi: 10.1186/s12934-024-02412-2.

DOI:10.1186/s12934-024-02412-2

PMID:38790055

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11127416/

Abstract

BACKGROUND

Azo dyes represent a common textile dye preferred for its high stability on fabrics in various harsh conditions. Although these dyes pose high-risk levels for all biological forms, fungal laccase is known as a green catalyst for its ability to oxidize numerous dyes.

METHODS

Trichoderma isolates were identified and tested for laccase production. Laccase production was optimized using Plackett-Burman Design. Laccase molecular weight and the kinetic properties of the enzyme, including K and V pH, temperature, and ionic strength, were detected. Azo dye removal efficiency by laccase enzyme was detected for Congo red, methylene blue, and methyl orange.

RESULTS

Eight out of nine Trichoderma isolates were laccase producers. Laccase production efficiency was optimized by the superior strain T. harzianum PP389612, increasing production from 1.6 to 2.89 U/ml. In SDS-PAGE, purified laccases appear as a single protein band with a molecular weight of 41.00 kDa. K and V values were 146.12 μmol guaiacol and 3.82 μmol guaiacol/min. Its activity was stable in the pH range of 5-7, with an optimum temperature range of 40 to 50 °C, optimum ionic strength of 50 mM NaCl, and thermostability properties up to 90 °C. The decolorization efficiency of laccase was increased by increasing the time and reached its maximum after 72 h. The highest efficiency was achieved in Congo red decolorization, which reached 99% after 72 h, followed by methylene blue at 72%, while methyl orange decolorization efficiency was 68.5%.

CONCLUSION

Trichoderma laccase can be used as an effective natural bio-agent for dye removal because it is stable and removes colors very well.

摘要

背景

偶氮染料是一种常用的纺织染料，因其在各种恶劣条件下对织物具有高稳定性而受到青睐。尽管这些染料对所有生物形式都构成高风险，但真菌漆酶因其能够氧化多种染料而被视为一种绿色催化剂。

方法

鉴定并测试了 Trichoderma 分离株的漆酶生产能力。使用 Plackett-Burman 设计对漆酶生产进行了优化。检测了漆酶的分子量和酶的动力学特性，包括 K 和 V pH、温度和离子强度。检测了漆酶酶对刚果红、亚甲基蓝和甲基橙的偶氮染料去除效率。

结果

9 株 Trichoderma 分离株中有 8 株是漆酶产生菌。通过优势菌株 T. harzianum PP389612 优化漆酶生产效率，使产量从 1.6 提高到 2.89 U/ml。在 SDS-PAGE 中，纯化的漆酶呈现出单一的蛋白质带，分子量为 41.00 kDa。K 和 V 值分别为 146.12 μmol 愈创木酚和 3.82 μmol 愈创木酚/分钟。其活性在 pH 5-7 范围内稳定，最适温度范围为 40-50°C，最适离子强度为 50 mM NaCl，热稳定性高达 90°C。通过增加时间，漆酶的脱色效率增加，并在 72 小时后达到最大值。刚果红的脱色效率最高，72 小时后达到 99%，其次是亚甲基蓝，达到 72%，而甲基橙的脱色效率为 68.5%。