薏米多肽-钙、锌螯合物制备及结构表征
Preparation and Structural Characterization of Coix Seed Polypeptide-Calcium-Zinc Chelate
胡香莲 1郦萍 1周柳莎 1俞瑜媛 1徐海星 1施永清1
作者信息
- 1. 浙江工商大学食品与生物工程学院 杭州 310018
- 折叠
摘要
目的:制备1种薏米多肽-钙、锌螯合物并研究其结构.方法:利用枯草芽孢杆菌和嗜热链球菌混合发酵薏米,采用葡聚糖凝胶Sephadex G-15、反相高效液相色谱(RP-HPLC)分离纯化薏米发酵液得到薏米多肽(CSP),Tricine-十二烷基硫酸钠-聚丙烯酰胺凝胶(Tricine-SDS-PAGE)电泳检测其分子质量.以锌螯合率和钙螯合率为指标,在单因素实验的基础上,采用响应面试验优化薏米多肽-钙、锌螯合物(CSP-Ca-Zn)的制备工艺,采用紫外光谱、红外光谱和荧光光谱表征其结构.结果:薏米发酵液经Sephadex G-15分离得到4个峰,其中A3的钙、锌螯合率最高.RP-HPLC分离A3得到的CSP纯度达93.91％,其分子质量约7.8 ku.CSP与Ca-Zn螯合的最佳制备工艺为:CSP与钙、锌质量比4.4∶1,钙、锌质量比1∶1,pH 3.7,在此条件下,锌螯合率、钙螯合率分别达到52.63％和63.79％.CSP与钙、锌螯合的主要位点是氨基氮、羧酸基,其空间结构也发生改变.结论:所确定的螯合工艺条件使CSP同时螯合钙、锌两种金属,为薏米多肽新产品的开发提供了技术参考,为制作食源性有机钙、锌补充剂提供了新思路.
Abstract
This paper's main aim has been to prepare an polypeptide-calcium-zinc chelate from coix seed and to inves-tigate its structure.Bacillus subtilis and Streptococcus thermophilus were used for fermentation of coix seed(CS).Polypeptide from coix seed(CSP)was separated and purified by gel filtration chromatography and reversed-phase high performance liquid chromatography(RP-HPLC).The molecular weight of CSP was detected by Tricine-sodium dodecyl sulfate-polyacrylamide gel(Tricine-SDS-PAGE)electrophoresis.Taking the chelation rate of zinc and calcium as indica-tors,single factor and response surface methodology were used to determine optimum preparation conditions of coix seed polypeptide-calcium-zinc(CSP-Ca-Zn)chelate.Ultraviolet spectra,fourier infrared and fluorescent light spectra were ap-plied to predict the structure of CSP-Ca-Zn.The results showed that four components were separated by Sephadex G-15.Among them,A3 showed the highest the chelating capacity of calcium and zinc.A3 was separated by RP-HPLC.The purity of CSP was as high as 93.91％,and its molecular mass was around 7.8 ku.The optimal chelate conditions were determined as follows:mass ratio of CSP to Ca-Zn 4.4:1,mass ratio of calcium to zinc 1:1,and pH 3.7.Under these conditions,the chelation rates of zinc and calcium were 52.63％and 63.79％,respectively.The principal binding sites of CSP with calcium and zinc were amino nitrogen and carboxylic acid group,and its spatial structure was also changed.This optimal chelate conditions could obtain CSP-Ca-Zn,which has provided technical reference for the devel-opment of new products of coix seed peptide and a new idea for food-borne organic calcium and zinc supplements.
关键词
薏米/发酵/纯化/肽-钙、锌螯合物/结构Key words
coix seed/fermentation/purification/peptide-calcium-zinc chelate/structure引用本文复制引用
出版年
2024
NETL
NSTL
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