螺旋藻 CPCC-695 漆酶的生化特征及其在雌酮降解中的作用

IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Neha Sami, Bushra Afzal, Durdana Yasin, Tasneem Fatma
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引用次数: 0

摘要

通过水生系统在食物链中添加雌酮等外源性内分泌干扰化合物(EDCs),会改变基因组或非基因组途径,从而扰乱类固醇的生物合成和代谢,最终导致各种疾病。因此,迫切需要对这些化合物进行生物修复,以防止其在环境中的添加和持续存在。事实证明,酶降解是一种安全且不产生有毒产物的 "盔甲骑士"。具有降解酚类和非酚类底物潜力的多铜氧化酶(E.C. 1.10.3.2 苯二醇:氧氧化还原酶)--漆酶最近受到了关注。本研究对漆酶进行了纯化、表征,并将其用于研究雌酮降解。培养滤液(粗漆酶)经浓缩后用冷丙酮沉淀,并用三羟甲基氨基甲烷缓冲液(50 mM)透析,得到四倍的部分纯化形式,产率为 45.56%,比活度为 204.14 U/mg,在 250-300 纳米波长处有一个单峰。根据 SDS-PAGE 估计,部分纯化的漆酶约为 80 kDa,以 ABTS 为底物。粗制和部分纯化的漆酶都在 pH 值为 3.0、温度为 40 °C、ABTS 为 4 mM 的条件下显示出最大活性。粗制和部分纯化的漆酶的动力学常数(Km、Vmax)分别为 0.83 mM;494.31 mM/min 和 0.58 mM;480.54 mM/min。硫酸铁和叠氮化钠对漆酶的抑制作用最大。粗提纯和部分提纯的漆酶降解效率分别为 87.55% 和 91.35%。螺旋藻 CPCC-695 漆酶具有高效的雌酮降解能力,因此有望用于 EDCs 的生物修复。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Biochemical Characterization of Laccase from Spirulina CPCC-695 and Their Role in Estrone Degradation

Biochemical Characterization of Laccase from Spirulina CPCC-695 and Their Role in Estrone Degradation

The addition of exogenous endocrine disrupting compounds (EDCs) like estrone, in the food chain through the aquatic system, disrupts steroid biosynthesis and metabolism by altering either the genomic or non-genomic pathway that eventually results in various diseases. Thus, bioremediation of these compounds is urgently required to prevent their addition and persistence in the environment. Enzymatic degradation has proven to be a knight in shining armour as it is safe and generates no toxic products. The multicopper oxidases (E.C. 1.10.3.2 benzenediol: oxygen oxidoreductase), laccase with the potential to degrade both phenolic and non-phenolic substrates has recently gained attention. In this study, the laccase was purified, characterized, and used to study estrone degradation. The culture filtrate (crude laccase) was concentrated and precipitated using cold-acetone and dialyzed against tris buffer (50 mM) giving a four-fold partially purified form, with 45.56% yield and 204.14 U/mg as specific activity and a single peak at 250–300 nm. The partially purified laccase was approximately 80 kDa as estimated by SDS-PAGE preferred ABTS as substrate. Both crude and partially purified laccase showed maximum activity at pH 3.0, 40 °C, and 4 mM ABTS. Kinetic constants (Km, Vmax) of crude and partially purified laccase were found to be 0.83 mM; 494.31 mM/min, and 0.58 mM; 480.54 mM/min respectively. Iron sulphate and sodium azide inhibited laccase maximally. Crude and partially purified laccase degradation efficiency was 87.55 and 91.35% respectively. Spirulina CPCC-695 laccase with efficient estrone degradation ability renders them promising candidates for EDCs bioremediation.

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来源期刊
The Protein Journal
The Protein Journal 生物-生化与分子生物学
CiteScore
5.20
自引率
0.00%
发文量
57
审稿时长
12 months
期刊介绍: The Protein Journal (formerly the Journal of Protein Chemistry) publishes original research work on all aspects of proteins and peptides. These include studies concerned with covalent or three-dimensional structure determination (X-ray, NMR, cryoEM, EPR/ESR, optical methods, etc.), computational aspects of protein structure and function, protein folding and misfolding, assembly, genetics, evolution, proteomics, molecular biology, protein engineering, protein nanotechnology, protein purification and analysis and peptide synthesis, as well as the elucidation and interpretation of the molecular bases of biological activities of proteins and peptides. We accept original research papers, reviews, mini-reviews, hypotheses, opinion papers, and letters to the editor.
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