木瓜籽肽-锌螯合物的制备、结构表征及稳定性研究

doi:10.3969/j.issn.0253-2417.2023.05.010

林产化学与工业 ›› 2023, Vol. 43 ›› Issue (5): 73-80.doi: 10.3969/j.issn.0253-2417.2023.05.010

木瓜籽肽-锌螯合物的制备、结构表征及稳定性研究

邓叶俊, 黄立新(), 张彩虹, 谢普军

中国林业科学研究院林产化学工业研究所; 江苏省生物质能源与材料重点实验室; 国家林业和草原局林产化学工程重点实验室; 林木生物质低碳高效利用国家工程研究中心; 江苏省林业资源高效加工利用协同创新中心, 江苏南京 210042

收稿日期:2022-07-12 出版日期:2023-10-28 发布日期:2023-10-27
通讯作者: 黄立新 E-mail:l_x_huang@163.com
作者简介:黄立新, 研究员, 博士生导师, 研究领域为天然资源化学利用; E-mail: l_x_huang@163.com
邓叶俊(1993—)，男，安徽宣城人，博士，主要从事林源食品深加工研究工作
基金资助:
国家自然科学基金资助项目(32171732)

Preparation, Structural Characterization and Stability Study of Chinese Quince Seed Peptide-Zinc Chelate

Yejun DENG, Lixin HUANG(), Caihong ZHANG, Pujun XIE

Institute of Chemical Industry of Forest Products, CAF; Key Lab. of Biomass Energy and Material, Jiangsu Province; Key Lab. of Chemical Engineering of Forest Products, National Forestry and Grassland Administration; National Engineering Research Center of Low-Carbon Processing and Utilization of Forest Biomass; Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing 210042, China

Received:2022-07-12 Online:2023-10-28 Published:2023-10-27
Contact: Lixin HUANG E-mail:l_x_huang@163.com

摘要/Abstract

摘要：

为提高Zn²⁺稳定性及生物利用度, 以木瓜籽为原料, 通过酶水解制备木瓜籽肽(CQSP), 经超滤获得5个不同分子质量范围的多肽组分, 并制备CQSP-Zn螯合物。考察了蛋白酶种类和多肽分子质量对Zn²⁺螯合率的影响, 运用氨基酸组成分析、红外光谱(FT-IR)、紫外-可见光谱(UV-Vis)、荧光光谱对螯合物结构特征进行表征, 并探究了CQSP-Zn在不同pH值和消化阶段的溶解度。研究结果表明：碱性蛋白酶水解的分子质量＜1 000 u的CQSP对Zn²⁺螯合率最高, 为83.26%。红外光谱、紫外光谱及荧光光谱分析表明：CQSP与Zn²⁺间产生了相互作用, CQSP中的—NH、CH₂、C=O、C—N、—COOH可能参与了对Zn²⁺的螯合。相对于无机ZnSO₄, CQSP-Zn在pH值2.0~8.0及模拟胃肠道消化过程中具有更高的溶解度, pH值为8.0时, CQSP-Zn的Zn²⁺溶解度为73.85%, 达到ZnSO₄的11.19倍；在pH值为7.5的模拟肠液中, 消化第0 min时CQSP-Zn的Zn²⁺溶解度为65.86%, 显著高于ZnSO₄的16.24%, 表明CQSP-Zn具有作为Zn²⁺增补剂的潜力。

关键词: 肽, 螯合物, 模拟消化, 生物利用度

Abstract:

In order to improve the stability and bioavailability of Zn²⁺, the Chinese quince seed peptide(CQSP) was prepared by enzymatic hydrolysis using Chinese quince seed as raw material. Five peptide fractions with different molecular weight ranges were obtained by ultrafiltration, and they were applied for preparation of CQSP-Zn chelate. The effects of protease types and peptide molecular weight on the chelation rate of Zn²⁺ were investigated. The amino acids composition analysis, Fourier transform infrared(FT-IR) spectroscopy, ultraviolet-visible(UV-Vis) spectroscopy and fluorescent spectroscopy were used to identify the structural characteristics of the chelate, and the solubility of CQSP-Zn at different pH values as well as digestion stages were also studied. The results indicated that the CQSP with the molecular weight less than 1 000 u hydrolyzed by alkaline protease presented the highest Zn²⁺ chelating rate(83.26%). The analysis of FT-IR spectroscopy, UV-Vis spectroscopy and fluorescent spectroscopy revealed the interaction between CQSP and Zn²⁺, and the -NH, CH₂, C=O, C-N, -COOH in CQSP might participate in the chelating of Zn²⁺. Compared with the inorganic ZnSO₄, CQSP-Zn showed higher solubility at pH 2.0-8.0 and simulated gastrointestinal digestion procedure. When the pH value was 8.0, the Zn²⁺ solubility of CQSP-Zn was 73.85%, which was 11.19 times higher than that of ZnSO₄. In the simulated intestinal fluid with a pH value of 7.5, the Zn²⁺ solubility of CQSP-Zn was 65.86% at 0 minute of digestion, significantly higher than ZnSO₄(16.24%), demonstrating potential application of CQSP-Zn as Zn²⁺ supplement.

Key words: peptide, chelate, simulate digestion, bioavailability

中图分类号:

TQ35
TS202.1

邓叶俊, 黄立新, 张彩虹, 谢普军. 木瓜籽肽-锌螯合物的制备、结构表征及稳定性研究[J]. 林产化学与工业, 2023, 43(5): 73-80.

Yejun DENG, Lixin HUANG, Caihong ZHANG, Pujun XIE. Preparation, Structural Characterization and Stability Study of Chinese Quince Seed Peptide-Zinc Chelate[J]. Chemistry and Industry of Forest Products, 2023, 43(5): 73-80.

图/表 7

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	条件 conditions	Zn²⁺螯合率¹⁾/% Zn²⁺ chelating rate
蛋白酶种类 protease species	碱性蛋白酶alkaline protease	70.84±1.747^a
	木瓜蛋白酶papain	38.98±0.671^b
	胰蛋白酶trypsin	37.37±1.528^b
	菠萝蛋白酶bromelain	36.99±1.026^b
	风味蛋白酶flavourzyme	24.01±1.275^c
肽分子质量 molecular weight of peptides	≤1 000 u	83.26±1.384^a
	>1 000-3 000 u	75.45±1.482^b
	>3 000-5 000 u	72.56±1.110^b
	>5 000-10 000 u	64.35±0.825^c
	＞10 000 u	49.67±0.909^d

氨基酸种类 type of amino acids	CQSP/%	CQSP-Zn/%	氨基酸种类 type of amino acids	CQSP/%	CQSP-Zn/%
天冬氨酸Asp	10.46	12.37	异亮氨酸Ile	6.56	4.27
苏氨酸Thr	2.35	2.03	亮氨酸Leu	7.20	4.06
丝氨酸Ser	3.49	3.74	酪氨酸Tyr	2.55	1.06
谷氨酸Glu	21.88	23.45	苯丙氨酸Phe	4.40	2.82
甘氨酸Gly	5.25	5.71	组氨酸His	4.80	6.74
丙氨酸Ala	3.13	2.13	赖氨酸Lys	1.90	3.41
胱氨酸Cys	0.86	0.69	精氨酸Arg	9.65	14.63
缬氨酸Val	4.35	3.53	脯氨酸Pro	6.61	5.58
蛋氨酸Met	4.56	3.83

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木瓜籽肽-锌螯合物的制备、结构表征及稳定性研究

Preparation, Structural Characterization and Stability Study of Chinese Quince Seed Peptide-Zinc Chelate

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