食品与发酵工业 >

2022 , Vol. 48 >Issue 7: 146 - 153

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.029457

研究报告

木薯叶片多肽的制备与抗氧化功能研究

  • 张作达 ,
  • 王琴飞 ,
  • 吴若娜 ,
  • 牛晓磊 ,
  • 张振文
展开
  • 1(海南大学 热带作物学院,海南 海口,570228)
    2(中国热带农业科学院 热带作物品种资源研究所,海南 海口,571101)
硕士研究生(牛晓磊副教授和张振文研究员为共同通信作者,E-mail: ninterxll@126.com; scuta96@163.com)

收稿日期: 2021-09-25

  修回日期: 2021-11-16

  网络出版日期: 2022-04-27

基金资助

国家重点研发计划项目(2081YFF0213505);国家木薯产业技术体系项目(CARS-11-HNZZW);科技部对发展中国家援助项目(KY202002005 )

收起

Preparation and antioxidant function of polypeptide from cassava leaf

  • ZHANG Zuoda ,
  • WANG Qinfei ,
  • WU Ruona ,
  • NIU Xiaolei ,
  • ZHANG Zhenwen
Expand
  • 1(College of Tropical Crops,Hainan University,Haikou 570228,China)
    2(Institute of Tropical Crop Variety Resources,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China)

Received date: 2021-09-25

  Revised date: 2021-11-16

  Online published: 2022-04-27

Fold

摘要

以木薯叶片为材料,采用碱溶酸沉法对木薯叶片蛋白质进行提取,通过正交法优化提取工艺,采用响应面法优化3种蛋白酶酶解条件,并对酶解所得多肽进行抗氧化功能验证。结果表明,木薯叶片蛋白质最佳提取条件为:温度60 ℃、碱溶pH 11.0、提取时间2 h;酸沉pH 4.6,每克叶片鲜样的蛋白质最高得率为11.73%,所得蛋白质的纯度为92.57%。响应曲面分析结果表明,利用碱性蛋白酶、中性蛋白酶和胰蛋白酶酶解木薯叶片蛋白质,其酶解最佳条件分别为:酶解温度36.74、35.00、36.45 ℃,pH值8.76、6.31、8.00,反应时间2.44、2.18、3.82 h,其中胰蛋白酶多肽得率最高,为41.89%。酶解所得多肽溶液中游离氨基酸含量均极低,仅检测到酪氨酸,其质量浓度为9.90 μg/mL。抗氧化功能分析发现,中性蛋白酶酶解所得多肽抗氧化活性均较高,在其多肽质量浓度为1.5 mg/mL时,其DPPH自由基清除率、·OH清除率和总还原力分别为91.83%、79.3%、0.558。因此,中性蛋白酶酶解所得多肽抗氧化活性最强,最适合制备木薯叶抗氧化肽。

关键词： 木薯叶片; 酶解; 多肽; 抗氧化; 蛋白质

本文引用格式

张作达 , 王琴飞 , 吴若娜 , 牛晓磊 , 张振文 . 木薯叶片多肽的制备与抗氧化功能研究[J]. 食品与发酵工业, 2022 , 48(7) : 146 -153 . DOI: 10.13995/j.cnki.11-1802/ts.029457

Abstract

In this experiment, cassava leaf protein was extracted by alkali solution and acid precipitation, and the extraction process was optimized by orthogonal method. The hydrolysis conditions of three proteases were optimized by response surface methodology, and the antioxidant function of the peptides was verified. The results showed that the optimum extraction conditions of cassava leaf protein were as follows: extraction with alkali soluble (pH 11.0) for 2 h at 60 ℃, acid precipitation at pH 4.6. Under these conditions, the highest yield of protein per gram of fresh leaves was 11.73% and the purity of protein was 92.57%. The results of response surface analysis showed that the optimum conditions for enzymatic hydrolysis of cassava leaf protein by alkaline protease, neutral protease and trypsin were as follows: enzymatic hydrolysis temperature 36.74 ℃, 35.00 ℃, 36.45 ℃, pH value 8.76, 6.31 and 8.00, reaction time 2.44 h, 2.18 h and 3.82 h, respectively. The yield of trypsin polypeptide was the highest(41.89%). The content of free amino acids in the peptide solution obtained by enzymatic hydrolysis was very low, only tyrosine could be detected, and its concentration was 9.90 μg/mL. The antioxidant function analysis showed that the antioxidant activities of the peptides hydrolyzed by neutral protease were high. When the polypeptide concentration was 1.5 mg/ml, the DPPH radical clearance, hydroxyl radical clearance and total reducing power were 91.83%, 79.3% and 0.558 4, respectively. Therefore, the peptides hydrolyzed by neutral protease had the strongest antioxidant activity. Among the three enzymes, neutral protease is the most suitable for the preparation of cassava leaf antioxidant peptides.

Key words： cassava leaf; enzymolysis; polypeptide; antioxidant; protein

参考文献

[1] 吴秋妃. 木薯叶提取液制备及其抗氧化作用研究[D].海口:海南大学,2019.
WU Q F.Study on the preparation and its antioxidant function of cassava leaf[D].Haikou:Hainan University,2019.
[2] LATIF S,MüLLER J.Potential of cassava leaves in human nutrition:A review[J].Trends in Food Science & Technology,2015,44(2):147-158.
[3] 蔡燕萍, 余晓婉,张庆春,等.水产品生物活性肽的研究进展[J].食品与发酵工业,2020,46(16):249-256.
CAI Y P,YU X W,ZHANG Q C,et al.Research progress of bioactive peptides in aquatic products[J].Food and Fermentation Industries,2020,46(16):249-256.
[4] GÖRGÜÇ A,GENÇDA E,YΙLMAZ F M.Bioactive peptides derived from plant origin by-products:Biological activities and techno-functional utilizations in food developments-A review[J].Food Research International,2020,136:109504.
[5] SARMADI B H,ISMAIL A.Antioxidative peptides from food proteins:A review[J].Peptides,2010,31(10):1 949-1 956.
[6] 殷丹, 雷磊,陈科力.江南卷柏提取物脱脂工艺优选[J].中国实验方剂学杂志,2014,20(24):24-26.
YIN D,LEI L,CHEN K L.Optimization of degreasing technology of extracts from Selaginella moelledorfii[J].Chinese Journal of Experimental Traditional Medical Formulae,2014,20(24):24-26.
[7] 张灿. 银杏活性多肽的制备及功能性多肽产品研究[D].南京:南京林业大学,2017.
ZHANG C.Preparation of ginkgo active peptide and the research of functional peptide product[D].Nanjing:Nanjing Forestry University,2017.
[8] 张妮. 南瓜籽蛋白及多肽制备的研究[D].武汉:武汉轻工大学,2019.
ZHANG N.Study on the preparation pumpkin seed protein and peptide[D].Wuhan:Wuhan Polytechnic University,2019.
[9] 李素云, 覃颖泉,谢冬梅,等.超声辅助酶解大米多肽的制备及其对酵母细胞增殖性影响[J].食品工业,2020,41(3):126-129.
LI S Y,QIN Y Q,XIE D M,et al.Research of preparation of rice peptides by ultrasonic-assisted enzymatic hydrolysis and effect on yeast cells' proliferation[J].The Food Industry,2020,41(3):126-129.
[10] 肖怀秋, 李玉珍,林亲录,等.响应面优化冷榨花生粕酶法制备多肽工艺的研究[J].中国粮油学报,2013,28(9):50-55.
XIAO H Q,LI Y Z,LIN Q L,et al.Response surface methodology optimization for peptide preparation from cold pressed peanut meal by enzymatic hydrolysis[J].Journal of the Chinese Cereals and Oils Association,2013,28(9):50-55.
[11] 王子秦. 大豆源生物活性肽生产工艺优化及ACE抑制活性确证[D].长春:吉林大学,2020.
WANG Z Q.Production process optimization and angiotensin I-converting enzyme activity confirmation of soybean-derived bioactive peptides[D].Changchun:Jilin University,2020.
[12] ZANG J H,XU Y S,XIA W S,et al.Correlations between microbiota succession and flavor formation during fermentation of Chinese low-salt fermented common carp(Cyprinus carpio L.) inoculated with mixed starter cultures[J].Food Microbiology,2020,90:103487.
[13] 李艳. 优化HPLC测定黑皮鸡枞菌中氨基酸含量样品处理的条件[J].食品研究与开发,2018,39(19):147-152.
LI Y.Optimization conditions of processed sample for the determination of amino acid content in Termitomyces spp.by HPLC[J].Food Research and Development,2018,39(19):147-152.
[14] 夏吉安, 黄凯,李森,等.绿豆抗氧化肽的酶法制备及其抗氧化活性[J].食品与生物技术学报,2020,39(10):40-47.
XIA J A,HUANG K,LI S,et al.Enzymatic preparation of mung bean antioxidant peptides and antioxidant activity investigation[J].Journal of Food Science and Biotechnology,2020,39(10):40-47.
[15] 袁艳超, 贾泽,赖爱兰,等.混菌发酵芝麻粕制备芝麻多肽及其体外抗氧化活性研究[J].基因组学与应用生物学,2018,37(5):2 068-2 073.
YUAN Y C,JIA Z,LAI A L,et al.Preparation of sesame polypeptide from sesame meal by mixed fermentation and its antioxidant activity in vitro[J].Genomics and Applied Biology,2018,37(5):2 068-2 073.
[16] 赵旭彤. 蓝莓加工废弃物中花青素提取纯化及抗氧化活性研究[D].长春:吉林大学,2015.
ZHAO X T.Research on extraction and purification of anthocyanins from blueberry processing byproducts and antioxidant activity[D].Changchun:Jilin University,2015.
[17] 马菲菲. 银杏种仁ACE抑制肽的分离纯化、鉴定及抑制机理的研究[D].合肥:合肥工业大学,2019.
MA F F.Isolation,purification,identification and inhibition mechanism of ACE inhibitory peptides from ginkgo seeds[D].Hefei:Hefei University of Technology,2019.
[18] 高泽汝, 刘昆仑,陈复生.碱法提取玉米胚芽蛋白的条件优化[J].食品工业,2020,41(10):5-8.
GAO Z R,LIU K L,CHEN F S.Optimization of extraction conditions for corn germ protein by alkali method[J].The Food Industry,2020,41(10):5-8.
[19] 毛晓英, 朱新荣,万银松,等.核桃蛋白的组成分析及分离提取工艺的优化[J].中国食品学报,2019,19(3):195-205.
MAO X Y,ZHU X R,WAN Y S,et al.The composition analyses of walnut protein and optimization of extraction process[J].Journal of Chinese Institute of Food Science and Technology,2019,19(3):195-205.
[20] 贾蕾,何慧,向极钎,等.碎米荠硒肽的制备及其体外抗氧化活性分析[J].食品工业科技,2022,43(6):206-213.
JIA L,HE H,XIANG J Q,et al.Preparation and in vitro antioxidant activity analysis of selenium peptide from shepherd's purse[J].Food Industry Science and Technology,2022,43(6):206-213.
[21] 吕凯波, 吕述权,朱家乐.不同提取方式对红花籽粕蛋白提取率及其功能特性的影响[J].食品科技,2020,45(7):249-254.
LYU K B,LYU S Q,ZHU J L.Effects of different extraction methods on extraction yield and functional properties of protein from safflower seed meal[J].Food Science and Technology,2020,45(7):249-254.
[22] 任为聪, 程建军,张智宇,等.不同改性方法对蛋白质溶解性的影响研究进展[J].中国粮油学报,2011,26(8):123-128.
REN W C,CHENG J J,ZHANG Z Y,et al.Study on different modification methods on solubility properties of protein[J].Journal of the Chinese Cereals and Oils Association,2011,26(8):123-128.
[23] 马利华, 秦卫东,陈学红.不同菌种制备大豆多肽的抗氧化活性比较研究[J].食品工业科技,2011,32(10):201-204.
MA L H,QIN W D,CHEN X H.Comparative study on antioxidative activities of soybean peptide produced by different microbiological strains[J].Science and Technology of Food Industry,2011,32(10):201-204.
[24] 赵梦鸽, 何慧,马爱民,等.蛋清源生物活性肽功能及其构效关系研究进展[J].华中农业大学学报,2020,39(5):167-175.
ZHAO M G,HE H,MA A M,et al.Biological functions and structure-activity relationships of bioactive peptides derived from egg white proteins:A review[J].Journal of Huazhong Agricultural University,2020,39(5):167-175.
[25] UNO S,KODAMA D,YUKAWA H,et al.Quantitative analysis of the relationship between structure and antioxidant activity of tripeptides[J].Journal of Peptide Science:an Official Publication of the European Peptide Society,2020,26(3):e3238.
[26] CHEN H M,MURAMOTO K,YAMAUCHI F.Structural analysis of antioxidative peptides from soybean β-conglycinin[J].Journal of Agricultural and Food Chemistry,1995,43(3):574-578.
[27] SUETSUNA K,UKEDA H,OCHI H.Isolation and characterization of free radical scavenging activities peptides derived from casein[J].The Journal of Nutritional Biochemistry,2000,11(3):128-131.
[28] TSUGE N,EIKAWA Y,NOMURA Y,et al.Antioxidative activity of peptides prepared by enzymatic hydrolysis of egg-white albumin[J].Journal of the Agricultural Chemical Society of Japan,1991,65(11):1 635-1 641.
[29] 孙国威. 大豆7S、11S蛋白酶解模拟分析及疏水性对抗氧化活性的影响[D].无锡:江南大学,2009.
SUN G W.Study on the simulative hydrolysis of soybean 7S and 11S protein and effects of hydrophobicity on antioxidant acitvities[D].Wuxi:Jiangnan University,2009.
[30] 张燕, 魏汝君,马中苏,等.蛋清源活性肽对过氧化氢诱导的HEK293细胞谷胱甘肽抗氧化系统的影响[J].中国食品学报,2020,20(2):79-86.
ZHANG Y,WEI R J,MA Z S,et al.The effect of bioactive peptides from egg white on glutathione anti-oxidant system against oxidative stress injury in human embryonic kidney 293 cells[J].Journal of Chinese Institute of Food Science and Technology,2020,20(2):79-86.
Options
文章导航

/

〈 〉

技术支持：北京玛格泰克科技发展有限公司