鄭志強(qiáng) 劉 晉 魏曉娟 郝利民 郭順堂
(1.中國(guó)農(nóng)業(yè)大學(xué)食品科學(xué)與營(yíng)養(yǎng)工程學(xué)院, 北京 100083; 2.中央軍委后勤保障部軍需裝備研究所, 北京 100010;3.首都機(jī)場(chǎng)出入境檢驗(yàn)檢疫局, 北京 101300)
加工條件及模擬胃腸消化對(duì)小麥肽抗氧化穩(wěn)定性的影響
鄭志強(qiáng)1,2劉 晉2魏曉娟3郝利民2郭順堂1
(1.中國(guó)農(nóng)業(yè)大學(xué)食品科學(xué)與營(yíng)養(yǎng)工程學(xué)院, 北京 100083; 2.中央軍委后勤保障部軍需裝備研究所, 北京 100010;3.首都機(jī)場(chǎng)出入境檢驗(yàn)檢疫局, 北京 101300)
小麥肽; 加工條件; 胃腸消化; 抗氧化穩(wěn)定性
小麥肽是以小麥蛋白為原料,經(jīng)蛋白酶酶解、離心、過(guò)濾、噴霧干燥等工藝制成,主要由不同分子質(zhì)量的多肽組成,是一種多肽混合物。小麥肽可作為一種安全可靠的新型食品原料添加應(yīng)用于食品加工中。近年來(lái),大豆肽、玉米肽等食品原料已被廣泛應(yīng)用于食品工業(yè)中,而小麥肽作為一種新資源食品原料應(yīng)用較少。小麥肽與大豆肽、玉米肽類似,同屬于植源性肽,可能也是一種良好的食品原料。因此,隨著小麥蛋白深加工利用的不斷深入,小麥肽具有非常廣闊的開發(fā)應(yīng)用前景。
小麥蛋白(蛋白質(zhì)質(zhì)量分?jǐn)?shù)82%),商丘華陽(yáng)生態(tài)農(nóng)業(yè)發(fā)展有限公司。
堿性蛋白酶(2.17×105U/mL)、風(fēng)味蛋白酶(3.09×104U/mL)、胃蛋白酶(6.78×104U/g)、胰蛋白酶(3.61×105U/g),諾維信(中國(guó))生物技術(shù)有限公司。
1,1-二苯基-2-三硝基苯肼(DPPH),Sigma公司;葡萄糖、蔗糖、檸檬酸、山梨酸鉀、苯甲酸鈉、NaCl、KCl、CaCl2、MgSO4、ZnSO4、CuSO4,國(guó)藥集團(tuán)化學(xué)試劑有限公司;其他所用試劑均為國(guó)產(chǎn)分析純。
LGJ-10型真空冷凍干燥機(jī),北京松源華興科技發(fā)展有限公司;P-2102UV型紫外分光光度計(jì),上海新嘉電子有限公司;CR22G型高速冷凍離心機(jī),日本日立公司;SHZ-B型水浴恒溫振蕩器,上海五相儀器儀表有限公司; PHS-3C型pH計(jì),上海理達(dá)儀器廠;BW3200S型電子天平,上海精密科學(xué)儀器有限公司。
1.3.1小麥肽制備
將小麥蛋白以0.112 g/mL的質(zhì)量濃度分散在去離子水中,在55℃的水浴鍋中保溫,調(diào)節(jié)溶液的pH值至8.5,加入2 200 U/g的堿性蛋白酶開始酶解,攪拌酶解4.3 h,酶解結(jié)束后迅速沸水滅酶10 min;后調(diào)節(jié)溫度至50℃、pH值至6.5,加入1 070 U/g的風(fēng)味蛋白酶進(jìn)行第2步酶解,攪拌酶解2.2 h;酶解過(guò)程中每20 min用1.0 mol/L NaOH或HCl調(diào)節(jié)溶液pH值使預(yù)設(shè)pH值保持不變,酶解結(jié)束后迅速沸水滅酶10 min,冷卻后3 000 r/min離心20 min,取上清液冷凍干燥,-20℃保存待用。
1.3.2自由基清除率測(cè)定
1.3.3溫度對(duì)小麥肽抗氧化穩(wěn)定性的影響
1.3.4pH值對(duì)小麥肽抗氧化穩(wěn)定性的影響
1.3.5食品原輔料對(duì)小麥肽抗氧化穩(wěn)定性的影響
1.3.6金屬離子對(duì)小麥肽抗氧化穩(wěn)定性的影響
1.3.7體外模擬胃腸消化對(duì)小麥肽抗氧化穩(wěn)定性的影響
人工胃液和人工腸液按照文獻(xiàn)[17]的方法進(jìn)行配制。
人工胃液配制:取濃度為1 mol/L稀鹽酸16.4 mL,加水約800 mL與胃蛋白酶10 g,搖勻后,加水稀釋至1 000 mL。
人工腸液配制:取磷酸二氫鉀6.8 g,加水500 mL使溶解,用0.1 mol/L NaOH溶液調(diào)節(jié)pH值至6.8;另取胰蛋白酶10 g,加水適量使其溶解,將兩液混合后,加水稀釋至1 000 mL。
體外模擬胃腸消化參照江慎華等[18]以及YOU等[19]的方法。
1.3.7.1人工胃液?jiǎn)为?dú)作用對(duì)小麥肽抗氧化穩(wěn)定性的影響
1.3.7.2人工腸液?jiǎn)为?dú)作用對(duì)小麥肽抗氧化穩(wěn)定性的影響
1.3.7.3模擬人體胃腸消化對(duì)小麥肽抗氧化穩(wěn)定性的影響
圖1 不同溫度處理對(duì)小麥肽DPPH自由基和·OH清除率的影響Fig.1 Effect of different temperature treatments on DPPH radical, · and ·OH scavenging activities of wheat peptide
圖2 不同pH處理對(duì)小麥肽DPPH自由基和·OH清除率的影響Fig.2 Effect of different pH values treatments on DPPH radical, · and ·OH scavenging activities of wheat peptide
2.3.1NaCl
圖3 NaCl對(duì)小麥肽DPPH自由基和·OH清除率的影響Fig.3 Effect of NaCl on DPPH radical, · and ·OH scavenging activities of wheat peptide
2.3.2糖類
圖4 不同糖類質(zhì)量分?jǐn)?shù)對(duì)小麥肽DPPH自由基和·OH清除率的影響Fig.4 Effects of different sugars on DPPH radical, · and ·OH scavenging activities of wheat peptide
2.3.3檸檬酸
圖5 檸檬酸質(zhì)量分?jǐn)?shù)對(duì)小麥肽DPPH自由基和·OH清除率的影響Fig.5 Effect of citric acid on DPPH radical, · and ·OH scavenging activities of wheat peptide
2.3.4防腐劑
圖6 不同防腐劑對(duì)小麥肽DPPH自由基和·OH清除率的影響Fig.6 Effect of different preservatives on DPPH radical, · and ·OH scavenging activities of wheat peptide
由圖 6 可知,3種自由基清除率隨著防腐劑山梨酸鉀、苯甲酸鈉質(zhì)量分?jǐn)?shù)的提高均沒(méi)有發(fā)生顯著變化,3種自由基清除率均保持在70%以上。這說(shuō)明食品加工中常用的防腐劑對(duì)小麥肽的抗氧化活性沒(méi)有影響,在國(guó)家標(biāo)準(zhǔn)規(guī)定的添加范圍內(nèi)添加山梨酸鉀和苯甲酸鈉不會(huì)改變小麥肽的抗氧化活性。
圖7 不同金屬離子對(duì)小麥肽DPPH自由基和·OH清除率的影響Fig.7 Effects of different metal ions on DPPH radical, · and ·OH scavenging activities of wheat peptide
2.5體外模擬胃腸消化對(duì)小麥肽抗氧化穩(wěn)定性的影響
圖8 體外模擬胃腸消化對(duì)小麥肽DPPH自由基和·OH清除率的影響Fig.8 Effect of simulated gastrointestinal digestion on DPPH radical, · and ·OH scavenging activities of wheat peptide
研究了不同加工條件及體外模擬胃腸消化對(duì)小麥肽抗氧化穩(wěn)定性的影響。研究結(jié)果顯示,對(duì)小麥肽分別進(jìn)行熱處理以及添加蔗糖、山梨酸鉀、苯甲酸鈉、K+、Ca2+、Mg2+對(duì)其抗氧化活性均沒(méi)有明顯影響;添加NaCl、葡萄糖、檸檬酸、Zn2+、Cu2+對(duì)其抗氧化活性均有一定提高作用;堿性條件下小麥肽抗氧化活性下降明顯;人工胃液?jiǎn)为?dú)消化有助于提高小麥肽的抗氧化活性,而人工腸液?jiǎn)为?dú)消化則導(dǎo)致其抗氧化活性明顯下降,人工胃液、人工腸液分步消化同樣致使小麥肽抗氧化活性有所下降,但仍能保持較高的抗氧化活性。不同加工條件及胃腸消化方式對(duì)小麥肽的抗氧化活性影響各異,因此,采取合理優(yōu)化的加工條件有利于避免小麥肽抗氧化活性的下降,本試驗(yàn)結(jié)果可為小麥肽在食品工業(yè)中的應(yīng)用提供理論參考。
1 WIESER H.Chemistry of gluten proteins[J].Food Microbiology,2007,24(2):115-119.
2 GIANIBELLI M C,LARROQUE O R,MACRITCHIE F,et al.Biochemical, genetic, and molecular characterization of wheat glutenin and its component subunits[J].Cereal Chemistry,2001,78(6):635-646.
3 DRAGO S R,GONZLEZ R J.Foaming properties of enzymatically hydrolysed wheat gluten[J].Innovative Food Science and Emerging Technologies,2000,1(4):269-273.
4 BOLLECKER S,VIROBEN G,POPINEAU Y,et al.Acid deamidation and enzymatic modification at pH10 of wheat gliadins: influence on their functional properties[J].Sciences Des Aliments,1990,10(2):343-356.
6 ZHU K X,ZHOU H M,QIAN H F.Antioxidant and free radical-scavenging activities of wheat germ protein hydrolysates (WGPH) prepared with alcalase[J].Process Biochemistry,2006,41(6):1296-1302.
7 WANG J S,ZHAO M M,ZHAO Q Z,et al.Antioxidant properties of papain hydrolysates of wheat gluten in different oxidation systems[J].Food Chemistry,2007,101(4):1658-1663.
8 MOTOI H,KODAMA T.Isolation and characterization of angiotensin I—converting enzyme inhibitory peptides from wheat gliadin hydrolysate[J].Food Nahrung,2003,47(5):354-358.
9 JIA J Q,MA H,ZHAO W R,et al.The use of ultrasound for enzymatic preparation of ACE-inhibitory peptides from wheat germ protein[J].Food Chemistry,2010,119(1):336-342.
10 THEWISSEN B G,PAULY A,CELUS I,et al.Inhibition of angiotensin I converting enzyme by wheat gliadin hydrolysates[J].Food Chemistry,2011,127(4):1653-1658.
11 FUKUDOME S I,JINSMAA Y,MATSUKAWA T,et al.Release of opioid peptides, gluten exorphins by the action of pancreatic elastase[J].FEBS Letters,1997,412(3):475-479.
12 CALZUOLA I,GIAVARINI F,SASSI P,et al.Short acidic peptides isolated from wheat sprout chromatin and involved in the control of cell proliferation. Characterization by infrared spectroscopy and mass spectrometry[J].Peptides,2005,26(11):2074-2085.
13 ZHU C Z,ZHANG W G,KANG Z L,et al.Stability of an antioxidant peptide extracted from Jinhua ham[J].Meat Science,2014,96(2):783-789.
14 LI X C,LIN J,GAO Y X,et al.Antioxidant activity and mechanism of Rhizoma Cimicifugae[J].Chemistry Central Journal,2012,6:Article140.
15 LI X C.Improved pyrogallol autoxidation method: a reliable and cheap superoxide-scavenging assay suitable for all antioxidants[J].Journal of Agricultural and Food Chemistry,2012,60(25):6418-6424.
16 HALLIWELL B,GUTTERIDGE J M C,ARUOMA O I.The deoxyribose method: a simple “test tube” assay for determination of rate constants for reactions of hydroxyl radicals[J].Analytical Biochemistry,1987,165(1):215-219.
17 國(guó)家藥典委員會(huì).中華人民共和國(guó)藥典(2010年版二部)[M].北京:中國(guó)醫(yī)藥科技出版社,2010.
18 江慎華,蔡志鵬,廖亮,等.丁香抗氧化活性物質(zhì)提取及人工胃腸液對(duì)其活性的影響[J/OL].農(nóng)業(yè)機(jī)械學(xué)報(bào),2012,43(7):149-155.http:∥www.j-csam.org/jcsam/ch/reader/view_abstract.aspx?flag=1&file_no=20120728&journal_id=jcsam.DOI:10.6041/j.issn.1000-1298.2012.07.028. JIANG Shenhua,CAI Zhipeng,LIAO Liang,et al.Extraction of antioxidants from clove and effect of artificial gastrointestinal juice immersion on its antioxidant properties[J/OL].Transactions of the Chinese Society for Agricultural Machinery,2012,43(7):149-155.(in Chinese)
19 YOU L,ZHAO M,REGENSTEIN J M,et al.Changes in the antioxidant activity of loach (Misgurnusanguillicaudatus) protein hydrolysates during a simulated gastrointestinal digestion[J].Food Chemistry,2010,120(3):810-816.
20 ARCAN I,YEMENICIOLU A.Antioxidant activity of protein extracts from heat-treated or thermally processed chickpeas and white beans[J].Food Chemistry,2007,103(2):301-312.
21 LIARDONY R,JOST R.Racemization of free and protein-bound amino acids in strong mineral acid[J].International Journal of Peptide and Protein Research,1981,18(5):500-505.
22 林松毅,郭洋,王瑩,等.蛋清抗氧化肽增效劑的優(yōu)化[J].華南理工大學(xué)學(xué)報(bào):自然科學(xué)版,2010,38(8):100-104. LIN Songyi,GUO Yang,WANG Ying,et al.Optimization of synergist of antioxidant peptide derived from egg white protein[J].Journal of South China University of Technology:Natural Science Edition,2010,38(8):100-104.(in Chinese)
23 GUéRARD F,SUMAYA-MARTINEZ M T.Antioxidant effects of protein hydrolysates in the reaction with glucose[J].Journal of the American Oil Chemists’ Society,2003,80(5):467-470.
24 DELGADO-ANDRADE C,MORALES F J,SEIQUER I,et al.Maillard reaction products profile and intake from Spanish typical dishes[J].Food Research International,2010,43(5):1304-1311.
25 BENJAKUL S,VISESSANGUAN W,PHONGKANPAI V,et al.Antioxidative activity of caramelisation products and their preventive effect on lipid oxidation in fish mince[J].Food Chemistry,2005,90(1-2):231-239.
26 SU G,ZHENG L,CUI C,et al.Characterization of antioxidant activity and volatile compounds of Maillard reaction products derived from different peptide fractions of peanut hydrolysate[J].Food Research International,2011,44(10):3250-3258.
27 王瑩.茶多酚的抗氧化和抑菌活性及其增效劑[J].生物學(xué)雜志,2007,24(5):54-56. WANG Ying.The antioxidation and antimicrobial activities of tea polyphenols and its increased reagents[J].Journal of Biology,2007,24(5):54-56.(in Chinese)
28 NKHILI E,BRAT P.Reexamination of the ORAC assay: effect of metal ions[J].Analytical and Bioanalytical Chemistry,2011,400(5):1451-1458.
29 RAJAPAKSE N,MENDIS E,JUNG W K,et al.Purification of a radical scavenging peptide from fermented mussel sauce and its antioxidant properties[J].Food Research International,2005,38(2):175-182.
30 杜枘宣.牛乳酪蛋白胃腸水解多肽檢測(cè)及其Caco-2細(xì)胞吸收模型建立[D].西安:陜西科技大學(xué),2016. DU Ruixuan.Detection of gastrointestinal hydrolysis from milk casein and its absorption through Caco-2 monolayer cell model [D].Xi’an:Shanxi University of Science and Technology,2016.(in Chinese)
31 陳亮,林峰,金鎮(zhèn)濤,等.玉米低聚肽穩(wěn)定性的研究[J].食品與發(fā)酵工業(yè),2009,35(12):61-65. CHEN Liang,LIN Feng,JIN Zhentao,et al.Study on stability of corn oligopeptide[J].Food and Fermentation Industries,2009,35(12):61-65.(in Chinese)
32 周存山,秦曉佩,余筱潔,等.綠鰭?cǎi)R面鲀魚皮蛋白抗氧化肽模擬胃腸消化制備[J/OL].農(nóng)業(yè)機(jī)械學(xué)報(bào),2015,46(8):211-216.http:∥www.j-csam.org/jcsam/ch/reader/view_abstract.aspx?flag=1&file_no=20150829&journal_id=jcsam.DOI:10.6041/j.issn.1000-1298.2015.08.029. ZHOU Cunshan,QIN Xiaopei,YU Xiaojie,et al.Antioxidant activity and characteristics of simulated gastrointestinal digestion hydrolysate from filefishNavodonseptentrionalisskin protein[J/OL].Transactions of the Chinese Society for Agricultural Machinery,2015,46(8):211-216.(in Chinese)
EffectsofProcessingConditionsandSimulatedGastrointestinalDigestiononAntioxidativeStabilityofWheatPeptide
ZHENG Zhiqiang1,2LIU Jin2WEI Xiaojuan3HAO Limin2GUO Shuntang1
(1.CollegeofFoodScienceandNutritionalEngineering,ChinaAgriculturalUniversity,Beijing100083,China2.TheQuartermasterEquipmentInstituteofLogisticSupportDepartment,CMC,Beijing100010,China3.CapitalAirportEntry-ExitInspectionandQuarantineBureau,Beijing101300,China)
wheat peptide; processing conditions; gastrointestinal digestion; antioxidative stability
TS210.9
A
1000-1298(2017)09-0330-07
10.6041/j.issn.1000-1298.2017.09.042
2017-05-16
2017-07-06
國(guó)家自然科學(xué)基金項(xiàng)目(31471582)和全軍后勤科研重點(diǎn)項(xiàng)目(BX116C007)
鄭志強(qiáng)(1984—),男,博士生,中央軍委后勤保障部軍需裝備研究所工程師,主要從事蛋白質(zhì)加工與利用研究,E-mail: zzq198408@126.com
郭順堂(1962—),男,教授,博士生導(dǎo)師,主要從事蛋白質(zhì)加工與利用研究,E-mail: shuntang@cau.edu.cn