Abstract
[Objective] To improve the physicochemical stability of walnut polypeptide liposomes (WP-L), delay walnut polypeptide (WP) release, and conduct bilayer polysaccharide modification on the WP-L using sodium alginate (SA) and chitosan (CS). [Methods] The effects of modifications by CS and SA in different mass fractions on the physicochemical properties of WP-L are investigated. Then, the structural morphology of the liposomes is characterized by transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) spectroscopy. Additionally, liposome stability under long-term storage conditions is examined. Finally, polypeptide release rates, release behavior, sustained-release characteristics, alongside bioactivity changes before and after digestion are evaluated via in vitro simulations of gastrointestinal digestion and release experiments. [Results] When the mass fractions of CS and SA are 0.6% and 0.5%, respectively, the encapsulation efficiency of WP by the prepared SA-CS-WP-L is up to 90.67%, with a ζ-potential of -17.10 mV. According to TEM observation, SA-CS-WP-L exhibits a spherical vesicle structure with a smooth surface and a core-shell structure. Changes in TEM image contrast indicate successful deposition of CS and SA onto the WP-L surface. In FTIR results, SA and CS form stable complexes on the WP-L surface through hydrophobic interactions and intermolecular hydrogen bonding. During the 19-day low-temperature storage experiment, SA-CS-WP-L still exhibits a high polypeptide encapsulation efficiency (70.77%) and stable physicochemical properties. In vitro gastrointestinal digestion simulations and polypeptide release assays demonstrate that SA and CS modifications effectively delay WP release during digestion while maintaining their sustained inhibitory efficacy against angiotensin-converting enzyme (ACE). Furthermore, WP release follows Fick diffusion behavior. [Conclusion] Bilayer surface modification of WP-L using SA and CS effectively protects WP and achieves sustained-release performance.
Publication Date
5-13-2026
First Page
1
Last Page
12
DOI
10.13652/j.spjx.1003.5788.2025.81155
Recommended Citation
Rui, CHEN; Fengjun, WANG; Guohong, ZHANG; Feng, JIN; and Hongchun, PENG
(2026)
"Preparation and characterization of walnut polypeptide liposomes with bilayer polysaccharide modification,"
Food and Machinery: Vol. 42:
Iss.
3, Article 1.
DOI: 10.13652/j.spjx.1003.5788.2025.81155
Available at:
https://www.ifoodmm.cn/journal/vol42/iss3/1
References
[1] 王宸轩,吴丹,张素榕,等.核桃粕固态发酵酱油的制备及抗氧化性分析 [J].食品与机械,2023,39(5):182-185,202.WANG C X,WU D,ZHANG S R,et al.Preparation of solid fermented soy sauce in walnut dregs and analysis of antioxidant properties [J].Food & Machinery,2023,39(5):182-185,202.
[2] YANG L K,ZHU W Z,DING Y Y,et al.New insights into altering the structures and improving the functional properties of walnut protein:synergistic high-pressure homogenization/limited enzymatic hydrolysis [J].Innovative Food Science & Emerging Technologies,2025,102:104018.
[3] WANG Y X,TAN B,CHEN C,et al.The phenolic profile of walnut meal protein isolate and interaction of phenolics with walnut protein [J].Food Research International,2023,170:113042.
[4] 韦雅虹.酶解法制备核桃肽及其相关产品开发 [D].阿拉尔:塔里木大学,2024:11-12.WEI Y H.Enzymatic preparation of walnut peptide and related product development [D].Ala'er:Tarim University,2024:11-12.
[5] 倪策,曹天红,陈敏,等.核桃粕源抗氧化活性肽的酶解制备及活性分析 [J].食品与机械,2024,40(5):51-61.NI C,CAO T H,CHEN M,et al.Enzymatic hydrolysis preparation and activity analysis of antioxidant peptides derived from walnut dregs [J].Food & Machinery,2024,40(5):51-61.
[6] 王 琛.核 桃 蛋 白 源 ACE 抑 制 肽 的 分 离 纯 化 与 鉴 定 [D].北 京:北京林业大学,2020:11-33.|WANG C.Isolation,purification and identification of ACE inhibitory peptide from walnut protein [D].Beijing:Beijing Forestry University,2020:11-33.
[7] WANG M,SUN X Y,LUO W,et al.Characterization and analysis of antioxidant activity of walnut-derived pentapeptide PW5 via nuclear magnetic resonance spectroscopy [J].Food Chemistry,2021,339:128047.
[8] QIN W,KETNAWA S,OGAWA Y.Effect of digestive enzymes and pH on variation of bioavailability of green tea during simulated in vitro gastrointestinal digestion [J].Food Science and Human Wellness,2022,11(3):669-675.
[9] 尤婷婷,马永强,赵若冰,等.叶黄素纳米脂质体的制备及体外消化特性研究 [J].食品与机械,2025,41(12):28-36.YOU T T,MA Y Q,ZHAO R B,et al.Preparation and in vitro digestion characteristics of lutein nanoliposomes [J].Food & Machinery,2025,41(12):28-36.
[10] LUO Y X,WANG F L,YUAN X Y,et al.Walnut peptide loaded proliposomes with hydroxyapatite as a carrier:fabrication,environmental stability,and in vitro digestion attribute [J].Food Research International,2022,162(Pt B):112057.
[11] CAO S N,HAO J,WANG Y Z,et al.Chitosan-coated nanoliposomes for the enhanced stability of walnut angiotensin-converting enzyme (ACE) inhibitory peptide [J].Journal of Food Science,2023,88(5):2 130-2 140.
[12] WANG X Y,CAO Z X,SU J Y,et al.Preparation of sodium alginate and chitosan modified curcumin liposomes and study on the formation of protein corona [J].International Journal of Biological Macromolecules,2025,293:139392.
[13] TAN X Y,LIU Y,WU X X,et al.Layer-by-layer self-assembled liposomes prepared using sodium alginate and chitosan:insights into vesicle characteristics and physicochemical stability [J].Food Hydrocolloids,2024,149:109606.
[14] LIU W L,LIU J H,LIU W,et al.Improved physical and in vitro digestion stability of a polyelectrolyte delivery system based on layer-by-layer self-assembly alginate–chitosan-coated nanoliposomes [J].Journal of Agricultural and Food Chemistry,2013,61(17):4 133-4 144.
[15] 贺建华,鹿麟,邵纯君,等.福林酚法与考马斯亮蓝法测定甘露聚糖肽口服溶液中蛋白质含量的比较 [J].中国药师,2017,20(10):1 861-1 863.HE J H,LU L,SHAO C J,et al.Comparison of protein content determination respectively by folin-ciocalteu method and coomassie bril-liant blue binding method for mannatide oral solution [J].China Pharmacist,2017,20(10):1 861-1 863.
[16] 方顺翔.核桃多肽纳米脂质体制备、结构及稳定性研究 [D].北京:北京林业大学,2020:31.FANG S X.Production of nanoliposomes loaded with walnut protein hydrolysates:structure and stability [D].Beijing:Beijing Forestry University,2020:31.
[17] SARABANDI K,JAFARI S M.Effect of chitosan coating on the properties of nanoliposomes loaded with flaxseed-peptide fractions:stability during spray-drying [J].Food Chemistry,2020,310:125951.
[18] RAMEZANZADE L,HOSSEINI S F,NIKKHAH M.Biopolymer-coated nanoliposomes as carriers of rainbow trout skin-derived antioxidant peptides [J].Food Chemistry,2017,234:220-229.
[19] LIU W L,LIU W,YE A Q,et al.Environmental stress stability of microencapsules based on liposomes decorated with chitosan and sodium alginate [J].Food Chemistry,2016,196:396-404.
[20] BRODKORB A,EGGER L,ALMINGER M,et al.INFOGEST static in vitro simulation of gastrointestinal food digestion [J].Nature Protocols,2019,14(4):991-1 014.
[21] SHISHIR M R I,KARIM N,GOWD V,et al.Pectin-chitosan conjugated nanoliposome as a promising delivery system for neohesperidin:characterization,release behavior,cellular uptake,and antioxidant property [J].Food Hydrocolloids,2019,95:432-444.
[22] ARISE A K,ALASHI A M,NWACHUKWU I D,et al.Antioxidant activities of Bambara groundnut (Vigna subterranea) protein hydrolysates and their membrane ultrafiltration fractions [J].Food & Function,2016,7(5):2 431-2 437.
[23] OYAIZU M.Studies on product of browning reaction prepared from glucose amine [J].Japanese Journal of Nutrition,1986,44:307-315.
[24] LI Z Y,PAULSON A T,GILL T A.Encapsulation of bioactive salmon protein hydrolysates with chitosan-coated liposomes[J].Journal of Functional Foods,2015,19:733-743.
[25] MAZLOOMI S N,MAHOONAK A S,GHORBANI M,et al.Physicochemical properties of chitosan-coated nanoliposome loaded with orange seed protein hydrolysate [J].Journal of Food Engineering,2020,280:109976.
[26] CHUN J Y,CHOI M J,MIN S G,et al.Formation and stability of multiple-layered liposomes by layer-by-layer electrostatic deposition of biopolymers [J].Food Hydrocolloids,2013,30(1):249-257.
[27] SHISHIR M R I,KARIM N,XU Y,et al.Improving the physicochemical stability and functionality of nanoliposome using green polymer for the delivery of pelargonidin- 3-O-glucoside [J].Food Chemistry,2021,337:127654.
[28] CHAY S Y,TAN W K,SAARI N.Preparation and characterisation of nanoliposomes containing winged bean seeds bioactive peptides [J].Journal of Microencapsulation,2015,32(5):488-495.
[29] ZHOU F,XU T,ZHAO Y J,et al.Chitosan-coated liposomes as delivery systems for improving the stability and oral bioavailability of acteoside [J].Food Hydrocolloids,2018,83:17-24.
[30] JEON S,YOO C Y,PARK S N.Improved stability and skin permeability of sodium hyaluronate-chitosan multilayered liposomes by Layer-by-Layer electrostatic deposition for quercetin delivery [J].Colloids and Surfaces B,Biointerfaces,2015,129:7-14.
[31] KARIM N,SHISHIR M R I,CHEN W.Surface decoration of neohesperidin-loaded nanoliposome using chitosan and pectin for improving stability and controlled release [J].International Journal of Biological Macromolecules,2020,164:2 903-2 914.
[32] 刘 欣.维 生 素 C/β-胡 萝 卜 素 复 合 脂 质 体 的 制 备 及 性 质 研 究[D].广州:华南理工大学,2020:61.LIU X.Preparation and properties of vitamin C/beta-carotene co-encapsulation liposomes [D].Guangzhou:South China University of Technology,2020:61.
[33] 龚魁杰.花生纳米肽制备与吸收转运机制研究 [D].北京:中国农业科学院,2016:75-87.GONG K J.Study on preparation of peanut nano-peptide and in vitro absorbiton and transport mechanism [D].Beijing:Chinese Academy of Agricultural Sciences,2016:75-87.
[34] KHAN M A,YUE C,FANG Z,et al.Alginate/chitosan-coated zein nanoparticles for the delivery of resveratrol [J].Journal of Food Engineering,2019,258:45-53.
[35] LIU W L,TIAN M M,KONG Y Y,et al.Multilayered vitamin C nanoliposomes by self-assembly of alginate and chitosan:long-term stability and feasibility application in mandarin juice[J].LWT-Food Science and Technology,2017,75:608-615.
[36] GIBIS M,THELLMANN K,THONGKAEW C,et al.Interaction of polyphenols and multilayered liposomal-encapsulated grape seed extract with native and heat-treated proteins [J].Food Hydrocolloids,2014,41:119-131.
[37] NAM J,VANDERLICK T K,BEALES P A.Formation and dissolution of phospholipid domains with varying textures in hybrid lipo-polymersomes [J].Soft Matter,2012,8(30):7 982-7 988.
[38] SONG F F,CHEN J,ZHANG Z Q,et al.Preparation,characterization,and evaluation of flaxseed oil liposomes coated with chitosan and pea protein isolate hydrolysates [J].Food Chemistry,2023,404(Pt A):134547.
[39] NIU Z G,CONEJOS-SÁNCHEZ I,GRIFFIN B T,et al.Lipid-based nanocarriers for oral peptide delivery [J].Advanced Drug Delivery Reviews,2016,106:337-354.
[40] ANSARIFAR E,MOHEBBI M,SHAHIDI F,et al.Novel multilayer microcapsules based on soy protein isolate fibrils and high methoxyl pectin:production,characterization and release modeling [J].International Journal of Biological Macromolecules,2017,97:761-769.
[41] LATORRES J M,AQUINO S,ROCHA M,et al.Nanoencapsulation of white shrimp peptides in liposomes:characterization,stability,and influence on bioactive properties[J].Journal of Food Processing and Preservation,2021,45(7):e15591.
[42] LI Y,ZHANG Y C,CHAI Z,et al.Entrapment of an ACE inhibitory peptide into ferritin nanoparticles coated with sodium deoxycholate:improved chemical stability and intestinal absorption [J].LWT-Food Science and Technology,2021,147:111547.
[43] RAMEZANZADE L,HOSSEINI S F,AKBARI-ADERGANI B,et al.Cross-linked chitosan-coated liposomes for encapsulation of fish-derived peptide [J].LWT-Food Science and Technology,2021,150:112057.
[44] 杨雪.基于胃肠消化的大分子大米降血压肽酶法制备及其过程原位实时监测 [D].镇江:江苏大学,2018:63.YANG X.The preparation of macromolecular rice hypotensive peptide basing on gastrointestinal digestion and in situ real-time monitoring of its enzymatic process [D].Zhenjiang:Jiangsu University,2018:63.
[45] MCCLEMENTS D J.Food hydrocolloids:application as functional ingredients to control lipid digestion and bioavailability [J].Food Hydrocolloids,2021,111:106404.
