WANG Jing,ZHANG Shaonan,LIU Zhaojun,et al.Extraction and Characterization of Lectin From Oudemansiella furfuracea[J].Northern Horticulture,2020,44(11):124-132.[doi:10.11937/bfyy.20193912]
鳞柄小奥德蘑凝集素的提取及性质研究
- Title:
- Extraction and Characterization of Lectin From Oudemansiella furfuracea
- Keywords:
- Oudemansiella furfuracea; lectin; double dilution method; agglutinability; hemagglutinating activity; sugarbinding specificity
- 文献标志码:
- A
- 摘要:
- 以鳞柄小奥德蘑粉为试材，采用磷酸缓冲溶液浸提凝集素，倍比稀释法测定其热稳定性、耐盐性、糖特异性和对多种血红细胞的凝集能力；以凝血活力为指标，采用单因素试验和正交实验优化了鳞柄小奥德蘑凝集素的提取工艺，以期研究鳞柄小奥德蘑凝集素的最佳提取工艺及其凝血活性。结果表明：鳞柄小奥德蘑凝集素可凝集兔、鸡、羊和人的血红细胞，对兔血红细胞的凝集能力最强，凝集能力无物种专一性和血型专一性；具有一定的耐热性和耐盐性，80 ℃及以上温度处理可显著降低其凝集能力，且随着处理时间的延长，凝集能力逐渐降低，80 ℃处理50 min或90 ℃处理10 min，凝集能力消失；含0~0.8 mol?L-1 NaCl的0.01 mol?L-1磷酸缓冲溶液对其凝集能力无显著性影响；特异性糖为N-乙酰-D-半乳糖胺。鳞柄小奥德蘑凝集素的最佳提取工艺为浸提液种类0.01 mol?L-1磷酸缓冲溶液，料液比1∶15 g?mL-1，浸提时间2 h，浸提液pH 7.0。
- Abstract:
- Phosphate buffered solution was used to extract lectin from Oudemansiella furfuracea and double dilution method was carried out to determine thermal stability，salinity tolerance，sugarbinding specificity and agglutinability of erythrocytes，respectively.Single factor experiment and orthogonal experiment were designed to optimize the extraction condition of lectin using hemagglutinating activity as evaluation index to investigate the extraction condition and some characteristics oflectin from Oudemansiella furfuracea.The results showed that lectin from Oudemansiella furfuracea could agglutinate all types of erythrocytes，including rabbit，chick，sheep and human erythrocytes without species specificity or blood type specificity of human，and its agglutinability of rabbit erythrocytes was strongest.Lectin from Oudemansiella furfuracea had a certain thermal stability and salinity tolerance.its agglutinability would not be damaged until the temperature increased to 80 ℃ and effected by phosphate buffered solution containing 0-0.8 mol?L-1 NaCl．Specific sugar of the lectin was N-acetyl-β-D-galactosamine.The optimal extraction condition was obtained as follows，extraction buffer was 0.01 mol?L-1 phosphate buffered solution，the ratio of material to liquid was 1∶15 g?mL-1，the extraction time was 2 hours,pH of extraction buffer was 7.0.
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备注/Memo
第一作者简介：王晶（1983-），女，山东东平人，博士，副教授，现主要从事生物活性物质等研究工作。E-mail：oucflora@163.com.基金项目：河北省自然科学基金青年科学基金资助项目（C2018408059）；河北省高等学校科学技术研究重点资助项目（ZD2019068）；大学生创新创业训练计划资助项目（201910100005）；廊坊市细胞工程与应用研究重点实验室资助项目（201903601）。收稿日期：2020-01-13