Improvement of the Purification Process for Earthworm DNase and Cytotoxicity Analysis

IF 1.9 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

The Protein Journal Pub Date : 2026-09-16 DOI:10.1007/s10930-026-10353-5

Zhiqiang Zhu, Xiuwei Wang, Zhen Gua

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引用次数: 0

Abstract

Objectives

Deoxyribonuclease (DNase) is widely used in treating obstructed purulent sputum drainage caused by bronchiectasis, lung abscess, cystic fibrosis, and atelectasis and inhibits biofilm formation.

Methods

This study optimized the earthworm DNase extraction process. Tissue homogenates were prepared using mechanical homogenization, followed by isoelectric precipitation and acetone precipitation. Crude earthworm extracts (EWE) were purified by ion exchange chromatography. DNase activity was evaluated by Bradford assay and UV absorption method.

Results

Method 1 took 42–45 h, while method 2 took 15–18 h. The specific activity of EWE1 was 677.48 ± 52.84 U/mg. The specific activity of EWE2 was 622.03 ± 49.76 U/mg. Following weak cation exchange chromatography and heparin affinity chromatography, the enzyme activity of the extract was 4443.33 ± 266.59 U/mg. The minimum working concentration was 1.88 µg/mL.

Conclusion

The improved extraction process markedly reduced the preparation time and enhanced DNase specific activity of earthworm extract.

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蚯蚓dna酶纯化工艺改进及细胞毒性分析。

目的：脱氧核糖核酸酶（脱氧核糖核酸酶）广泛用于治疗支气管扩张、肺脓肿、囊性纤维化、肺不张等引起的脓性痰引流阻塞，并抑制生物膜的形成。方法：对蚯蚓dna酶的提取工艺进行优化。组织匀浆采用机械匀浆，然后等电沉淀和丙酮沉淀。采用离子交换色谱法对蚯蚓粗提物进行纯化。采用Bradford法和紫外吸收法测定dna酶活性。结果：方法1作用42 ~ 45 h，方法2作用15 ~ 18 h， EWE1比活性为677.48±52.84 U/mg。EWE2的比活性为622.03±49.76 U/mg。经弱阳离子交换层析和肝素亲和层析，其酶活为4443.33±266.59 U/mg。最低工作浓度为1.88µg/mL。结论：改进的提取工艺显著缩短了提取时间，提高了蚯蚓提取物的dna酶比活性。

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来源期刊

The Protein Journal 生物-生化与分子生物学

CiteScore

5.20

自引率

0.00%

发文量

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.