高活性酪蛋白磷酸肽螯合锌的制备及表征
发布时间：2018-09-19 11:25
【摘要】：锌对人体健康非常重要,锌缺乏导致多种疾病,而酪蛋白磷酸肽是一种具有促进锌离子吸收的活性多肽,可以螯合成酪蛋白磷酸肽螯合锌,开发作为一种补锌剂。目前市场多见的补锌制剂如硫酸锌等老一代无机锌盐,相比酪蛋白磷酸肽螯合锌等新一代多肽螯合补锌剂,虽然成本低,来源易获得,但刺激度高,毒性大,生物利用度低,酪蛋白磷酸肽螯合锌在活性和安全性均优于前一代锌补剂。因此本研究以具有锌螯合活性的酪蛋白磷酸肽(CPPs)为研究对象,可开发作为新一代有机锌补剂的材料。以碱性蛋白酶酶解得到的酪蛋白磷酸肽为原料,经Q Sepharose FF阴离子交换色谱与RP-HPLC(反相高效色谱)色谱法两步分离后得到的高锌螯合活性的CPPs,在pH 7.0,CPPs与锌源物料比4:1,60℃,80min反应条件下进行螯合。经SDS-PAGE电泳检测,该肽段分子量10k Da,螯合活性达88.68μg/mg,与纯化前粗肽相比,多肽含量提高3.6倍,锌结合量提高了10倍。以上述两部纯化后得到的高锌螯合活性的酪蛋白磷酸肽为原料,对比螯合前后,经氨基酸组成分析、紫外光谱、红外光谱以及固体核磁共振13C谱、1H谱、31P谱解析,结果表明,螯合前后CPPs的结构及氨基酸组成发生了明显变化,其中的磷酸基团参与了螯合,其主要结合位点是-COOH、-NH2及P-OH中的-OH。在体外模拟胃肠道消化环境,考察酪蛋白磷酸肽螯合锌的生物利用度(溶解度以及透析度),对比葡萄糖酸锌和ZnSO4的生物利用度,酪蛋白磷酸肽螯合锌则高于其两者;酪蛋白磷酸肽螯合锌释放Zn2+的速率慢于Zn SO4和葡萄糖酸锌,能减少对胃的刺激;膳食纤维和Fe2+过量的存在会影响酪蛋白磷酸肽螯合锌的生物利用度。酪蛋白磷酸肽螯合锌在模拟胃肠道消化过程中具有生物稳定性,可以作为可靠的高生物利用度的锌补剂。
[Abstract]:Zinc is very important to human health. Zinc deficiency leads to many diseases. Casein phosphopeptide is a kind of active peptide which can promote the absorption of zinc ion. It can be chelated to zinc casein phosphopeptide chelate and developed as a zinc supplement agent. Compared with the new generation of polypeptide chelating zinc reagents, such as zinc sulfate and zinc sulfate, the most common zinc supplements in the market are of low cost and easy to obtain, but they have high stimulation degree, high toxicity and low bioavailability. The activity and safety of casein phosphopeptide chelate zinc were better than that of previous zinc supplement. Therefore, in this study, casein phosphopeptide (CPPs) with zinc chelating activity can be used as a new generation of organic zinc supplement material. Casein phosphopeptide obtained by enzymatic hydrolysis of alkaline protease was used as raw material. Q Sepharose FF anion exchange chromatography (Q Sepharose FF) and RP-HPLC (reversed-phase high performance chromatography) chromatography were used to chelate high zinc chelating CPPs, under the reaction conditions of pH 7.0% and zinc source material ratio of 4: 1 ~ 60 鈩，
本文编号：2249996
[Abstract]:Zinc is very important to human health. Zinc deficiency leads to many diseases. Casein phosphopeptide is a kind of active peptide which can promote the absorption of zinc ion. It can be chelated to zinc casein phosphopeptide chelate and developed as a zinc supplement agent. Compared with the new generation of polypeptide chelating zinc reagents, such as zinc sulfate and zinc sulfate, the most common zinc supplements in the market are of low cost and easy to obtain, but they have high stimulation degree, high toxicity and low bioavailability. The activity and safety of casein phosphopeptide chelate zinc were better than that of previous zinc supplement. Therefore, in this study, casein phosphopeptide (CPPs) with zinc chelating activity can be used as a new generation of organic zinc supplement material. Casein phosphopeptide obtained by enzymatic hydrolysis of alkaline protease was used as raw material. Q Sepharose FF anion exchange chromatography (Q Sepharose FF) and RP-HPLC (reversed-phase high performance chromatography) chromatography were used to chelate high zinc chelating CPPs, under the reaction conditions of pH 7.0% and zinc source material ratio of 4: 1 ~ 60 鈩，
本文编号：2249996
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