刘蕾1,2 魏玉西1△ 刘淇2△ 赵玲1 王凌燕1.坛紫菜(Porphyra Haitanensis)中抑菌活性肽的制备与初步纯化[J].,2011,11(3):476-481 |
坛紫菜(Porphyra Haitanensis)中抑菌活性肽的制备与初步纯化 |
Preliminary Purification and Influencing Factors of AntimicrobialPolypeptides from Porphyra Haitannsis |
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DOI: |
中文关键词: 坛紫菜 水溶性蛋白 抗菌肽 纯化 影响因素 |
英文关键词: Porphyra haitanensis Water-soluble protein Antimicrobial polypeptide Purification Influencing factor |
基金项目:农业部公益性行业专项(200903030) |
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中文摘要: |
目的:提取坛紫菜中水溶性蛋白,并对其进行初步纯化和抑菌活性影响因素研究。方法:坛紫菜水溶性蛋白胃蛋白酶在
37℃、pH1.8条件下酶解3h,再经超滤、Bio-Gel P-10 和DEAE Sephadex A-50 层析纯化步骤得到一定分子量范围的多肽混合物。
采用平板打孔法和对金黄色葡萄球菌的抑制作用,跟踪测定活性多肽纯化过程及其活性影响因素。结果:SDS-PAGE测定结果表
明该抗菌多肽分子量介于43.0KD~66.2KD之间。它对金黄色葡萄球菌生长的抑制作用随着温度的升高逐渐减弱,5%EDTA、5%
柠檬酸、5%维生素C及25%二甲基亚砜对它的抑菌活性有协同作用,而5%维生素E则对它的抑菌活性有拮抗作用。结论:从坛
子菜水溶性蛋白中初步纯化得到的分子量介于43.0KD~66.2KD 的多肽,对金黄色葡萄球菌的生长有明显地抑制作用,并且它的
抑菌活性受到温度,部分有机酸和维生素的影响。 |
英文摘要: |
Objective: Extracting and purifying the water-soluble protein from Porphyra haitanensis, and to investigate the inhibitory
activity on Staphyloccocus aureu. Methods: The water-soluble protein was hydrolysated by pepsin at 37℃ and pH1.8 for 3h. The
hydrolysates were purified sequentially by ultrafiltration, Bio-Gel P-10 and DEAE Sephadex A-50 chromatography. The molecular
weight of antibacterial polypeptides was detcted by SDS-PAGE. The antimicrobial activity of every fraction obtained during purification
procedures was detected by plat stiletto method. The inhibitory effect on Staphyloccocus aureu, and the activity influencing factors of
polypeptides were detected. Results: The molecular weight f antibacterial polypeptides was between 43.0KD and 66.2KD. The antibacterial
peptides' antimicrobial activity to Staphyloccocus aureus was affected by temperature intensively. On the other hand, the synergistic
effects were observed in all combination of DSA-1 + 5%EDTA, DSA-1+ 5% citric acid, DSA-1 + 5%Vc or DSA-1+ 25%DMSO, and antagonistic
effects were observed in combination of DSA-1 + 5%VE+ 25%DMSO. Conclusion: The antibacterial polypeptides was purified
from water-soluble protein of Porphyra haitanensis, which molecular weight was between 43.0KD and 66.2KD. The antibacterial
activity of the antibacterial polypeptides was affected by temperature, some organic acid and some vitamine. |
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