三种方法提取的澳洲坚果分离蛋白的功能性质与构象的关系

摘要/Abstract

摘要： 本文采用碱提法（MAPI）、三（羟甲基）氨基甲烷（Tris（hydroxymethyl）aminomethane, Tris-HCl）提取法（MTPI）及氯化钠提取法（MSPI）从澳洲坚果脱脂粉中提取得到三种分离蛋白并对其功能和构象进行比较研究。研究发现，在酸性pH范围内MSPI比MAPI和MTPI有更高的溶解性，而在碱性pH范围内恰好相反。MSPI比MAPI和MTPI具有更高的乳化活性指数。MAPI、MTPI和MSPI的蛋白含量分别为88.9%，80.3%和86.1%。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gel electrophoresis, SDS-PAGE）和氨基酸分析表明，三种分离蛋白具有相似的条带分布和氨基酸组成，但它们的条带强度和氨基酸数量不同。此外，与MAPI和MTPI相比，MSPI具有较高的表面疏水性、荧光强度和较小的粒度。场发射扫描电子显微镜（field emission scanning electron microscope, SEM）结果表明，MSPI比MAPI和MTPI具有更多的由不同大小的球状结构组成的颗粒。

关键词: 澳洲坚果, 分离蛋白, 功能特性, 构象

Abstract: Functional and conformational properties of three protein isolates from defatted macadamia nut kernel flour prepared by alkaline (MAPI), Tris-HCl (MTPI), or sodium chloride (MSPI) extraction methods were investigated. MSPI were found to have higher solubility than MAPI and MTPI in the acid pH regions, while as for alkaline pH regions, MSPI showed lower solubility than that of MAPI and MTPI . MSPI has higher emulsifying activity index (246.88±34.23 m2/g) than that of MAPI and MTPI. The protein contents of MAPI, MTPI and MSPI were 88.9%, 80.3% and 86.1%, respectively. The SDS-PAGE and amino acid analysis showed that three protein isolates had similar protein band and amino acid patterns, but different band intensity and amino acid amount. Furthermore, MSPI showed higher surface hydrophobicity (730.86), fluorescence intensity and smaller particle size than that of MAPI and MTPI. And SEM results revealed MSPI had more heterogeneous particles consisting of globular structure in different sizes compared with MAPI and MTPI. Overall, the difference of conformation of proteins induced by three prepared methods may contribute to their diversity of functional properties.

Key words: Macadamia, protein isolates, functional properties, conformation

中图分类号:

TS255.6

陈豪钟俊桢黄宗兰钟业俊刘成梅. 三种方法提取的澳洲坚果分离蛋白的功能性质与构象的关系[J]. 食品科学, 0, (): 0-0.

CHEN HAO yejun zhong. Comparison of Functional Properties of Macadamia (Macadamia integrifolia) Proteins Prepared by Three Different Methods Related to Their Conformation[J]. FOOD SCIENCE, 0, (): 0-0.

参考文献

[1] VENKATACHALAM M, SATHE S K. Chemical composition of selected edible nut seeds[J]. Journal of Agricultural and Food Chemistry, 2006, 54(13): 4705-4714. DOI:10.1021/jf0606959.
[2] CURD J D, WERGOWSKE G, DOBBS J C, et al. Serum lipid effects of a high-monounsaturated fat diet based on macadamia nuts[J]. Archives of Internal Medicine, 2000, 160(8): 1154-1158. DOI:10.1001/archinte.160.8.1154.
[3] KAIJSER A, DUTTA P, SAVAGE G. Oxidative stability and lipid composition of macadamia nuts grown in New Zealand[J]. Food Chemistry, 2000, 71(1): 67-70. DOI:10.1016/S0308-8146(00)00132-1.
[4] MAGUIRE L S, O'SULLIVAN S M, GALVIN K, et al. Fatty acid profile, tocopherol, squalene and phytosterol content of walnuts, peanuts, hazelnuts and the macadamia nut[J]. International Journal of Food Sciences and Nutrition, 2004, 55(3):171-178. DOI:10.1080/09637480410001725175.
[5] ALBERTSON P L, GIOVANNI C D, COCKSEDGE R H, et al. Inducing biochemical changes to simulate after-roast darkening in macademia kernel[J]. Australian Journal of Experimental Agriculture, 2005, 45(10):1315-1323. DOI:10.1071/EA04176.
[6] WALL M M, GENTRY T S. Carbohydrate composition and color development during drying and roasting of macadamia nuts (Macadamia integrifolia)[J]. LWT - Food Science and Technology, 2007, 40(4):587-593. DOI:10.1016/j.lwt.2006.03.015.
[7] STEPHENSON R A, GALLAGHER E C. Effects of temperature during latter stages of nut development on growth and quality of macadamia nuts[J]. Scientia Horticulturae, 1986, 30(3):219-225. DOI:10.1016/0304-4238(86)90100-7.
[8] OGUNWOLU S O, HENSHAW F O, MOCK H P, et al. Production of protein concentrate and isolate produced from cashew (Anacardum occidentale L.) nut[J]. African Journal of Food, Agriculture, Nutrition and Development, 10, 10(5):2501-2514. DOI:10.4314/ajfand.v10i5.56334.
[9] SIWAPORN J, JUTHAMAS H, KANITHA T. Chemical compositions, functional properties, and microstructure of defatted macadamia flours[J]. Food Chemistry, 2008, 110(1):23-30. DOI:10.1016/j.foodchem.2008.01.050.
[10] BORA P S, RIBEIRO D. Note: influence of pH on the extraction yield and functional properties of macadamia (Macadamia integrofolia) protein isolates[J]. Food Science and Technology International, 2004, 10(4):263-267. DOI:10.1177/1082013204045779.
[11] LIU C M, ZHONG J Z, LIU W, et al. Relationship between functional properties and aggregation changes of whey protein induced by high pressure microfluidization[J]. Journal of Food Science, 2011, 76(4):341-347. DOI: 10.1111/j.1750-3841.2011.02134.x.
[12] BRADFORD M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding [J]. Analytical Biochemistry, 1976, 72(s 1–2):248–254. DOI:10.1016/003-2697(76)90527-3.
[13] ZHAO G L, YAN L, ZHAO M M, et al. Enzymatic hydrolysis and their effects on conformational and functional properties of peanut protein isolate[J]. Food Chemistry, 2011, 127(4):1438-1443. DOI:10.1016/j.foodchem.2011.01.046
[14] LAEMMLI U K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J]. Nature, 1970, 227(5259):680-685. DOI:10.1038/22768a0.
[15] LIU Y, ZHAO G, ZHAO M, et al. Improvement of functional properties of peanut isolate by conjugation with dextran through Maillard reaction[J]. Food Chemistry, 2012, 131(3):901-906. DOI: 10.1016/j.foodchem.2011.09.074.
[16] KATO A, NAKAI S. Hydrophobicity determined by a fluorescence probe method and its correlation with surface properties of proteins[J]. Biochimica et Biophysica Acta (BBA)-Protein Structure, 1980, 624(1):13-20. DOI: 10.1016/0005-2795(80)90220-2
[17] OGUNWOLU S O, HENSHAW F O, MOCK H P, et al. Functional properties of protein concentrates and isolates produced from cashew (Anacardum occidentale L.) nut[J]. Food Chemistry, 2009, 115(3):852-858. DOI:10.1016/j.foodchem.2009.09.007.
[18] PINTO RAMOS C M, BORA P S. Functionality of succinylated brazil nut (Bertholletia excelsa HBK) kernel globulin[J]. Plant Foods for Human Nutrition, 2005, 60(1):1-6. DOI:10.1007/s11130-005-2533-0.
[19] SZE-TAO K W C, SATHE S K. Walnuts (julans regia L): proximate composition, protein solubility, protein amino acid composition and protein in vitro digestibility[J]. Journal of the Science of Food and Agriculture, 2000, 80(9):1393-1401. DOI:10.1002/1097-0010(200007)80:9<1393::AID-JSFA653>3.0.CO;2-F.
[20] SWANSON B G. Pea and lentil protein extraction and functionality[J]. Journal of the American Oil Chemists' Society, 1990, 67(5):276-280. DOI:10.1007/BF02539676.
[21] PAPALAMPROU E M, DOXASTAKIS G I, KIOSSEOGLOU V. Chickpea protein isolates obtained by wet extraction as emulsifying agents[J]. Journal of the Science of Food and Agriculture, 2010, 90(2):304–313. DOI: 10.1002/jsfa.3816.
[22] LIU L H, HUNG T V, BENNETT L. Extraction and characterization of chickpea (Cicer arietinum) albumin and globulin[J]. Journal of Food Science, 2008, 73(5):299-305. DOI:10.1111/j.1750-3841.2008.00773.x.
[23] ALUKO R E, MCINTOSH T. Polypeptide profile and functional properties of defatted meals and protein isolates of canola seeds[J]. Journal of the Science of Food and Agriculture, 2001, 81(4):391–396. DOI:10.1002/1097-0010(200103)81:4<391::AID-JSFA823>3.0.CO;2-S.
[24] YOSHIKO-STARK Y, WADA Y, WASCHE A. Chemical composition, functional properyies, and bioactivities of rapeseed protein isolates[J]. Food Chemistry, 2008, 107(1):32-39. DOI: 10.1016/j.foodchem.2007.07.061.
[25] JAMDAR S N, RAJALAKSHMI V, PEDNEKAR M D, et al. Influence of degree of hydrolysis on functional properties, antioxidant activity and ACE inhibitory activity of peanut protein hydrolysate[J]. Food Chemistry, 2010, 121(1):178-184. DOI: 10.1016/j.foodchem.2009.12.027.
[26] KARACA A C, LOW N, NICKERSON M. Emulsifying properties of canola and flaxseed protein isolates produced by isoelectric precipitation and salt extraction[J]. Food Research International, 2011, 44(9):2991-2998. DOI:10.1006/j.foodres.2011.07.009.
[27] KRAUDE J P, SCHULTZ M, DUDEK S. Effect of extraction conditions composition, surface activity and rheological of protein isolates from flaxseed (Linum usitativissimum L)[J]. Journal of the Science of Food & Agriculture, 2002, 82(9):970-976. DOI:10.1002/jsfa.1140.
[28] MOURE A, SINEIRO J, DOMINGUEZ H, et al. Functionality of oilseed protein products: A reviw. Food Research International, 2006, 39(9):945-963. DOI:10.1006/j.foodres.2006.07.002.
[29] AYDEMIR L Y, GOKBULUT A A, BARAN Y, et al. Bioactive, functional and edible film-forming properties of isolated hazelnut[J]. Food Hydrocolloids, 2014, 36:130-142. DOI:10.1016/j.foodhyd.2013.09.014.
[30] DU Y, JIANG Y, ZHU X, et al. Physicochemical and functional properties of the protein isolate and major fractions prepared from Akebia trifoliatea var. australis seed[J]. Food Chemistry, 2012, 133(3):923-929. DOI:10.1006/j.foodchem.2012.02.005.
[31] HORAX R, HETTIARACHCHY N, KANNAN A, et al. Protein extraction optimization, characterization, and functionalities of protein isolate from bitter melon (Momordica charantia) seed[J]. Food Chemistry, 2011, 124(2):545-550. DOI:10.1006/j.foodchem.2010.06.068.
[32] KARACA A C, LOW N, NICKERSON M. Emulsifying properties of chickpea, faba bean, lentil and pea proteins produced by isoelectric precipitation and salt extraction[J]. Food Research International, 2011, 44(9):2742-2750. DOI:10.1016/j.foodres.2011.06.012.
[33] BOUAOUINA H, DESRUMAUX A, LOISEL C, et al. Functional properties of whey proteins as affected by dynamic high-pressure treatment[J]. International Dairy Journal, 2006, 16(4):275-284. DOI:10.1016/j.idairyj.2005.05.004.