Scientia Agricultura Sinica ›› 2020, Vol. 53 ›› Issue (6): 1247-1255.doi: 10.3864/j.issn.0578-1752.2020.06.016

• FOOD SCIENCE AND ENGINEERING • Previous Articles     Next Articles

Wheat Gluten, Gliadins and Glutenin Content Changes During Germination Based on the Methods of R5 ELISA and RP-HPLC

HuiMin HU,XueFeng PAN,Heng YANG,Chen CHEN,YinJi CHEN()   

  1. College of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing 210023
  • Received:2019-07-30 Accepted:2019-10-30 Online:2020-03-16 Published:2020-04-09
  • Contact: YinJi CHEN E-mail:chenyinji@hotmail.com

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Abstract:

【Objective】 The current study was conducted to determine the dynamic changes of gluten, gliadins and glutenin contents in different germination states of wheat, so as to provide a scientific basis for the development of gluten-free food and utilization of germinated wheat. 【Method】Seven wheat grains with different germination states were obtained by controlling the germination conditions. Changes of composition of gliadins and glutenin were analyzed by sodium dedecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and changes of gluten, gliadins and glutenin subunits during wheat germination were further determined by enzyme-linked immunosorbent assay based on R5 antibody (R5 ELISA) and reversed-phase high-performance liquid chromatography (RP-HPLC). 【Result】The content of gluten, gliadins and glutenin subunits in wheat could be determined by R5 ELISA and RP-HPLC. Germination treatment had different effects on the allergic proteins and subunits mentioned above. The content of gluten changed little at the early stage of germination, but decreased significantly at the later stage. The relative content of ω-gliadins did not change significantly. During the germination process, the relative content of α-/β-gliadins was significantly reduced, with the percentage of untreated wheat seeds decreased from 41.85% to 31.51%-35.35% after germination (P＜0.01). The relative content of γ-gliadins increased significantly from 31.37% to 36.69%-39.02% after germination (P＜0.05). The relative content changes of high molecular weight glutenin subunit (HMW-GS) and low molecular weight glutenin subunit (LMW-GS) were not significant. HMW-GS decreased slightly from 8.66% (untreated group) to 5.94% (1/4 bud length) and then to 7.28% (bud length=grain length). LMW-GS increased slightly from 8.30% (untreated group) to 10.45% (bud length=grain length).【Conclusion】Both of the methods R5 ELISA and RP-HPLC could be used for quantitative analysis of wheat sensitized proteins. The content of allergenic protein in wheat decreased during germination. In particular, when the germinating bud was up to 1/2 long, the α-/β-gliadin which containing the most sensitive peptide decreased significantly. It was suggested that moderate germination treatment could reduce sensitization of wheat.

Key words: celiac disease, gliadin, glutenin, gluten, R5 ELISA, RP-HPLC

Fig. 1

Changes of wheat in different germination states A-G represent unsoaked, soaked, exposed, seed coat broken, bud length 1/4, bud length 1/2, bud length equals grain length. The same as below"

Fig. 2

SDS-PAGE of gliadins from wheat in different germination states"

Fig. 3

SDS-PAGE of glutenin from wheat in different germination states"

Fig. 4

Gluten changes during wheat germination determined by R5 ELISA The lowercase letters indicate significant differences at 0.05 level"

Fig. 5

RP-HPLC of gliadins and glutenin subunits from wheat in different germination states"

Table 1

Relative amounts of gliadins and glutenin from wheat in different germination state (%)"

亚基
Subunit		 发芽状态Germination state
A B C D E F G
ω 9.48±1.06a 9.81±0.40a 9.69±0.84a 11.30±1.00a 10.20±0.37a 12.03±0.47a 10.57±0.88a
α+β 41.85±0.63A 33.81±1.65B 34.36±1.83B 35.35±0.77B 33.19±1.35B 31.51±0.32B 33.52±0.77B
γ 31.37±0.76a 36.69±0.37b 39.02±1.04b 37.62±2.40b 38.70±2.27b 37.28±1.29b 36.91±1.10b
HMW 8.66±0.61a 8.98±1.33a 8.12±1.11a 6.32±1.15a 5.94±0.34a 8.00±0.20a 7.28±0.19a
LMW 8.30±0.74a 9.80±0.61a 8.81±0.40a 8.73±0.80a 10.13±1.03a 10.49±0.33a 10.45±0.43a
[1] LEE G J, KAO J Y . Recent advances in pediatric celiac disease. Expert Review of Gastroenterology & Hepatology, 2017,11(6):583-592.
[2] BOUZIAT R, HINTERLEITNER R, BROWN J J , et al. Reovirus infection triggers inflammatory responses to dietary antigens and development of celiac disease. Science, 2017,356(6333):44-50.
[3] HOLMES G K T, MUIRHEAD A . Epidemiology of coeliac disease in a single centre in Southern Derbyshire 1958-2014. BMJ Open Gastroenterology, 2017,4(1):e000137.
[4] CHO K, BEOM H R, JANG Y R, ALTENBACH S B, VENSEL W H, SIMON-BUSS A, LIM S H, KIM M G, LEE J Y . Proteomic profiling and epitope analysis of the complex α-, γ-, and ω-gliadin families in a commercial bread wheat. Frontiers in Plant Science, 2018,9:818.
[5] JIANG Y R, BEOM H R, ALTENBACH S B, LEE M K, LIM S H, LEE J Y . Improved method for reliable HMW-GS identification by RP-HPLC and SDS-PAGE in common wheat cultivars. Molecules, 2017,22(7):1055.
[6] WIESER H, KOEHLER P . The biochemical basis of celiac disease. Cereal Chemistry, 2008,85(1):1-13.
[7] KOGA S, BÖCKER U, WIESER H, KOEHLER P, UHLEN A K, MOLDESTAD A . Polymerisation of gluten proteins in developing wheat grain as affected by desiccation. Journal of Cereal Science, 2017,73:122-129.
[8] 赵艳丽 . 发芽小麦粉的利用研究[D]. 郑州: 河南工业大学, 2013.
ZHAO Y L . Study on the use of germinated wheat flour[D]. Zhengzhou: Henan University of Technology, 2013. (in Chinese)
[9] 董召荣, 柯建国, 马传喜 . 不同品种小麦籽粒发芽深度对其加工品质的影响. 南京农业大学学报, 2000,23(2):9-12.
DONG Z R, KE J G, MA C X , Effect of different germinating conditions on kernel qualities in common wheat. Journal of Nanjing Agricultural University, 2000,23(2):9-12. (in Chinese)
[10] OLAERTS H, ROYE C, DERDE L J, SINNAEVE G, MEZA W R, BODSON B, COURTIN C M . Impact of preharvest sprouting of wheat (Triticum aestivum) in the field on starch, protein, and arabinoxylan properties. Journal of Agricultural and Food Chemistry, 2016,64:8324-8332.
[11] Codex Committee of Methods of Analysis and Sampling. MAS/36 CRD/15. Report on the 36th session in Budapest, Hungary, 2015.
[12] DIAZ-AMIGO C, POPPING B . Accuracy of ELISA detection methods for gluten and reference materials: A realistic assessment. Journal of Agricultural and Food Chemistry, 2013,61(24):5681-5688.
[13] SCHERF K A, POMS R E . Recent developments in analytical methods for tracing gluten. Journal of Cereal Science, 2016,67:112-122.
[14] SHETTY R, VESTERGAARD M, JESSEN F, HÄGGLUND P, KNORR V, KOEHLER P, PRAKASH S H, HOBLEY T J . Discovery, cloning and characterisation of proline specific prolyl endopeptidase, a gluten degrading thermo-stable enzyme from, Sphaerobacter thermophiles. Enzyme and Microbial Technology, 2017,107:56-63
[15] CHO S W, KANG C S, KANG T G, CHO K M, PARK C S . Influence of different nitrogen application on flour properties,gluten properties by HPLC and end-use quality of Korean wheat. Journal of Integrative Agricultural, 2018,17(5):982-993.
[16] GIANIBELLI M C, ECHAIDE M, LARROQUE O R, CARRILLO J M, DUBCOVSKY J . Biochemical and molecular characterisation of Glu-1 loci in Argentinean wheat cultivars. Euphytica, 2002,128(1):61-73.
[17] ZHOU J X, LIU D M, DENG X, ZHEN S M, WANG Z M, ZHOU J W . Effects of water deficit on breadmaking quality and storage protein compositions in bread wheat (Triticum aestivum L.). Journal of the Science of Food and Agriculture, 2018,98:4357-4368.
[18] ŚWIECA M, DZIKI D, GAWLIK-DZIKI U . Starch and protein analysis of wheat bread enriched with phenolics-rich sprouted wheat flour. Food Chemistry, 2017,228:643-648.
[19] 刘可欣, 杨润强, 顾振新, 王沛 . 发芽对小麦营养和加工品质影响研究进展. 中国粮油学报, 2018,33(5):135-140.
LIU K X, YANG R Q, GU Z X, WANG P . Research progress for effect of germination on wheat nutrition and processing quality. Journal of Chinese Cereals and Oils Association, 2018,33(5):135-140. (in Chinese)
[20] 中华人民共和国国家标准. 小麦. GB 1351-2008.
中华人民共和国国家标准. Wheat. GB 1351-2008. (in Chinese)
[21] 张佳灵 . 芽萌动对小麦粉品质影响的研究[D]. 武汉: 武汉轻工大学, 2015.
ZHANG J L . Influence of germination on wheat flour quality[D]. Wuhan: Wuhan Polytechnic University, 2015. (in Chinese)
[22] 菅桂玲, 田建珍, 郑学玲, 刘翀, 韩小贤 . 发芽时间对不同系统小麦粉品质的影响. 粮食与饲料工业, 2015(3). doi: 10.7633/j.issn. 1003-6202.2015.03.003.
JIAN G L, TIAN J Z, ZHEN X L, LIU C, HAN X X . Impact of germination time on wheat flour quality in different milling streams. Cereal and Feed Industry, 2015(3). doi: 10.7633/j.issn.1003-6202. 2015.03.003. (in Chinese)
[23] BOUKID F, PRANDI B, BUHLER S, SFORZA S . Effectiveness of germination on protein hydrolysis as a way to reduce adverse reactions to wheat. Journal of Agricultural and Food Chemistry, 2017,65(45):9854-9860.
[24] 邱然, 杨文洲, 张春玲 . 萌发大麦种子内肽酶活力、醇溶蛋白及氨基酸含量变化. 食品与生物技术学报, 2007,26(5):48-52.
QIU R, YANG W Z, ZHANG C L . Changes in endopeptidase activity, content of hordein and free amino acids in barley seeds during germination. Journal of Food and Biotechnology, 2007,26(5):48-52. (in Chinese)
[25] PRANDI B, BENCIVENNI M, FACCINI A, TEDESCHI T, DOSSENA A, MARCHELLI R, GALAVERNA G, SFORZA S . Composition of peptide mixtures derived from simulated gastrointestinal digestion of prolamins from different wheat varieties. Journal of Cereal Science, 2012,56(2):223-231.
[26] BOUKID F, PRANDI B, SFORZA S, SAYAR R, SEO Y W, MEJRI M, YACOUBI I . Understanding the effects of genotype, growing year and breeding on Tunisian durum wheat allergenicity. 1. The Baker’s Asthma case. Journal of Agricultural and Food Chemistry, 2017,65(28):5831-5836.
[27] CICCOCIOPPO R, DI SABATINO A, CORAZZA G R . The immune recognition of gluten in coeliac disease. Clinical & Experimental Immunology, 2005,140(3):408-416.
[28] BIGIARINI L . Hydrolysis of gliadin during germination of wheat seeds. Journal of Plant Physiology, 1995,147(2):161-167.
[29] SINGH P, ARORA A, STRAND T A, LEFFLER D A, CATASSI C, GREEN P H, KELLY C P, AHUJA V, MAKHARIA G K . Global prevalence of celiac disease: Systematic review and meta-analysis. Clinical Gastroenterology and Hepatology, 2018,16(6):823-836.
[30] LIU E, DONG F, BARÓN A E, TAKI I, NORRIS J M, FROHENERT B I, HOFFENBERG E J, REWERS M . High incidence of celiac disease in a long-term study of adolescents with susceptibility genotypes. Gastroenterology, 2017,152(6):1329-1336.
[31] ABADIE V, SOLLID L M, BARREIRO L B, JABRI B . Integration of genetic and immunological insights into a model of celiac disease pathogenesis. Annual Review of Immunology, 2011,29:493-525.
[32] YUAN J L, ZHOU C Y, GAO J Y, LI J J, LU J, LI X, WANG X Z, TONG P, WU Z H, YANG A S, YAO Y H, NADIF S, SHU H, JIANG X, WU Y J, GILISSEN L, CHEN H B . Prevalence of celiac disease autoimmunity among adolescents and young adults in China. Clinical Gastroenterology and Hepatology, 2017,15(10):1572-1579.
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