食品与发酵工业 >

2024 , Vol. 50 >Issue 4: 197 - 203

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.034297

研究报告

蛋清多肽-铁螯合物的制备、表征及对Caco-2细胞的促增殖作用

  • 王晴晴 ,
  • 蔡为荣 ,
  • 汪玉玲 ,
  • 卓允允 ,
  • 黄伟 ,
  • 曹宇凡
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  • (安徽工程大学 生物与食品工程学院,安徽 芜湖,241000)
第一作者:硕士研究生(蔡为荣教授为通信作者,E-mail:caiweirong0781@163.com)

收稿日期: 2022-11-14

  修回日期: 2022-12-13

  网络出版日期: 2024-03-15

基金资助

安徽省重点研发项目(202004a06020022)

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Preparation, characterization, and proliferation by Caco-2 cells of egg white polypeptide-iron chelate

  • WANG Qingqing ,
  • CAI Weirong ,
  • WANG Yuling ,
  • ZHUO Yunyun ,
  • HUANG Wei ,
  • CAO Yufan
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  • (School of Biological and Food Engineering, Anhui Polytechnic University, Wuhu 241000, China)

Received date: 2022-11-14

  Revised date: 2022-12-13

  Online published: 2024-03-15

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摘要

为开发人体易吸收的铁补充剂,提高咸蛋清的附加值,该研究以铁螯合能力和水解度(degree of hydrolysis,DH)为指标,优化酶解条件,通过超滤和Sephadex G-15 凝胶柱分离纯化铁螯合能力高的蛋清多肽。采用扫描电镜、能谱、红外光谱和X射线衍射(X-ray diffraction,XRD)对蛋清多肽-铁螯合物结构进行表征。同时探究其对Caco-2细胞活力的影响。结果表明,最佳酶解条件为底物质量分数4%、酶添加量10%、pH 8、温度50 ℃时酶解8 h,所得产物的铁螯合能力为95.27%。蛋清粗肽经超滤和Sephadex G-15 分离纯化得到铁螯合能力最高的组分F4-4。扫描电镜、能谱、红外光谱和XRD结果表明蛋清多肽和Fe2+形成多肽-铁螯合物。细胞实验中,与蛋清多肽和硫酸亚铁相比,蛋清多肽-铁螯合物的安全性高,且有一定的促增殖作用。

关键词： 咸蛋清; 铁结合肽; 分离纯化; 结构表征; 细胞增殖

本文引用格式

王晴晴 , 蔡为荣 , 汪玉玲 , 卓允允 , 黄伟 , 曹宇凡 . 蛋清多肽-铁螯合物的制备、表征及对Caco-2细胞的促增殖作用[J]. 食品与发酵工业, 2024 , 50(4) : 197 -203 . DOI: 10.13995/j.cnki.11-1802/ts.034297

Abstract

This study aimed to produce iron supplements that can be easily absorbed by humans and increase the added value of salted egg white. The optimum enzymatic hydrolysis conditions were obtained based on the iron chelating capacity and degree of hydrolysis, and the polypeptides with high iron chelating capacity were separated and purified by ultrafiltration and Sephadex G-15 gel column. The structure of egg white polypeptide-iron chelate was characterized by scanning electron microscopy, energy spectrum, infrared spectroscopy, and X-ray diffraction (XRD). At the same time, this study also aimed to explore its effect on the cell activity of Caco-2. Results showed that the optimal conditions of enzymatic hydrolysis were that substrate concentration was 4%, enzyme addition was 10%, pH was 8, temperature was 50 ℃, and enzymatic hydrolysis time was 8 h. The iron chelating capacity of the product was 95.27%. F4-4 was the highest iron chelating capacity component, which was obtained from egg white polypeptide by ultrafiltration and Sephadex G-15. The results of the scanning electron microscope, energy spectrum, infrared spectrum, and XRD showed that the egg white polypeptide formed a chelate with Fe2+. In the cell experiment, compared with egg white polypeptide and ferrous sulfate, the egg white polypeptide-iron was safer and could have a certain role in promoting cell proliferation.

Key words： salted egg white; iron-binding peptide; separation and purification; structural characterization; cell proliferation

参考文献

[1] TAN T C, PHATTHANAWIBOON T, MAT EASA A.Quality, textural, and sensory properties of yellow alkaline noodles formulated with salted duck egg white[J].Journal of Food Quality, 2016, 39(4):342-350.
[2] 赵宁宁, 何慧, 胡琪, 等.脱盐咸鸭蛋清促钙吸收肽的制备及脱酰胺修饰[J].食品科学, 2014, 35(9):181-186.
ZHAO N N, HE H, HU Q, et al.Preparation and deamination of calcium absorption-promoting peptides from desalted duck egg white[J]. Food Science, 2014, 35(9):181-186.
[3] 曹丛丛. 阿胶促铁吸收的活性成分及其铁螯合物的补血作用研究[D].无锡:江南大学, 2021.
CAO C C. Study on the active components of Colla corii asini promoting iron absorption and hematopoietic effect of their iron chelates.Wuxi: Jiangnan University, 2021.
[4] 黄赛博. 带鱼蛋白多肽亚铁螯合物的制备及生物活性研究[D].舟山:浙江海洋大学, 2016.
HUANG S B. Study on preparation and biological activities of ferrous chelates including hairtail protein hydrolysates.Zhoushan: Zhejiang Ocean University, 2016.
[5] ZHANG Y J, DING X J, LI M Q.Preparation, characterization and in vitro stability of iron-chelating peptides from mung beans[J].Food Chemistry, 2021, 349:129101.
[6] GUO L D, HOU H, LI B F, et al.Preparation, isolation and identification of iron-chelating peptides derived from Alaska pollock skin[J].Process Biochemistry, 2013, 48(5-6):988-993.
[7] WU H H, LIU Z Y, ZHAO Y H, et al.Enzymatic preparation and characterization of iron-chelating peptides from anchovy (Engraulis japonicus) muscle protein[J].Food Research International, 2012, 48(2):435-441.
[8] LI B, HE H, SHI W, et al.Effect of duck egg white peptide-ferrous chelate on iron bioavailability in vivo and structure characterization[J].Journal of the Science of Food and Agriculture, 2019, 99(4):1834-1841.
[9] 刘闪, 刘良忠, 陈阳明, 等.白鲢鱼骨胶原低聚肽螯合钙的结构表征及其促细胞增殖作用[J].中国食品学报, 2016, 16(9):185-191.
LIU S, LIU L Z, CHEN Y M, et al.Structural characterization and promoting cell proliferation of collagen oligopeptide chelated calcium from silver carp fish bone[J].Journal of Chinese Institute of Food Science and Technology, 2016, 16(9):185-191.
[10] HUANG W, LAN Y Q, LIAO W W, et al.Preparation, characterization and biological activities of egg white peptides-calcium chelate[J].LWT, 2021, 149:112035.
[11] WANG Y H, CAI M, ZENG H, et al.Preparation, characterization and iron absorption by Caco-2 cells of the casein peptides-iron chelate[J].International Journal of Peptide Research and Therapeutics, 2022, 28(4):116.
[12] 崔春, 任娇艳, 赵谋明, 等.一种咸蛋清脱盐制备高起泡性蛋清粉的方法:中国, CN101878924B[P].2012-10-31.
CUI C, REN Y J, ZHAO M M, et al.A method for preparing high foaming egg white powder by desalting salted egg white:China, CN101878924B[P].2012-10-31.
[13] 李博, 侯焘, 陈恩民, 等.脱盐咸鸭蛋清肽-亚铁螯合物的制备及表征[J].食品工业科技, 2017, 38(16):172-177.
LI B, HOU T, CHEN E M, et al.Preparation and characterization of desalted duck egg white peptide-ferrous chelate[J].Science and Technology of Food Industry, 2017, 38(16):172-177.
[14] LIU Y, MA X Y, SI X X, et al.Preparation, characterization, and properties of wampee seed antioxidant peptides-iron chelate[J].Journal of Food Processing and Preservation, 2022, 46(5):e16513.
[15] REN Y, WU H, LI X F, et al.Purification and characterization of high antioxidant peptides from duck egg white protein hydrolysates[J].Biochemical and Biophysical Research Communications, 2014, 452(4):888-894.
[16] 庞忠莉. 牡蛎肽亚铁螯合物的制备及性质研究[D].广州:华南理工大学, 2020.
PANG Z L. Study on preparation and properties of oyster peptide ferrous chelate[D].Guangzhou:South China University of Technology, 2020.
[17] FANG S X, RUAN G R, HAO J, et al.Characterization and antioxidant properties of Manchurian walnut meal hydrolysates after calcium chelation[J].LWT-Food Science and Technology, 2020, 130:109632.
[18] ZHU S Y, ZHENG Y M, HE S, et al.Novel Zn-binding peptide isolated from soy protein hydrolysates:Purification, structure, and digestion[J].Journal of Agricultural and Food Chemistry, 2021, 69(1):483-490.
[19] 钱跃威, 徐瀚麟, 吕奇晏, 等.鳗鱼骨胶原肽钙螯合物的制备及其稳定性和Caco-2吸收特性[J].食品科学, 2020, 41(24):1-8.
QIAN Y W, XU H L, LYU Q Y, et al.Preparation and stability of calcium-chelating peptide and its absorption characteristics in Caco-2 cells[J].Food Science, 2020, 41(24):1-8.
[20] 谢丽平. 具有抗氧化、抗衰老活性的多肽筛选、分离纯化及结构鉴定[D].广州:华南理工大学, 2019.
XIE L P. Screening, separation and purification, structure identification of peptides with antioxidant and anti-aging effects. Guangzhou: South China University of Technology, 2019.
[21] JIANG H, ZHANG W T, CHEN F Y, et al.Purification of an iron-binding peptide from scad (Decapterus maruadsi) processing by-products and its effects on iron absorption by Caco-2 cells[J].Journal of Food Biochemistry, 2019, 43(7):e12876.
[22] 曹玉惠, 张娟娟, 王再扬, 等.牡蛎源类蛋白反应修饰肽的分离纯化及肽锌螯合物的结构表征[J].高等学校化学学报, 2018, 39(3):470-475.
CAO Y H, ZHANG J J, WANG Z Y, et al.Separation and identification of oyster peptide modified by plastein reaction and characterization of peptide-zinc complexes[J].Chemical Journal of Chinese Universities, 2018, 39(3):470-475.
[23] WANG L, DING Y Y, ZHANG X X, et al.Isolation of a novel calcium-binding peptide from wheat germ protein hydrolysates and the prediction for its mechanism of combination[J].Food Chemistry, 2018, 239:416-426.
[24] ATHIRA S, MANN B, SHARMA R, et al.Preparation and characterization of iron-chelating peptides from whey protein:An alternative approach for chemical iron fortification[J].Food Research International, 2021, 141:110133.
[25] HE W L, FENG Y, LI X L, et al.Availability and toxicity of Fe(II) and Fe(III) in Caco-2 cells[J].Journal of Zhejiang University SCIENCE B, 2008, 9(9):707-712.
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