乌鸡低聚肽铁的分离纯化及结构鉴定
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摘要
为了考察乌鸡低聚肽铁中肽段的结构,以乌鸡为原料,以氯化亚铁为铁源制备乌鸡低聚肽铁,对其总蛋白质质量分数、螯合率、得率进行测定,结果表明乌鸡低聚肽铁总蛋白质质量分数为(56.79±0.18)%,螯合率为(83.53±0.15)%,得率为(40.86±0.14)%。然后利用反相高效液相色谱(RP-HPLC)对其进行分离纯化,收集4个组分峰,利用四极杆飞行时间串联质谱仪(QTOF质谱仪)测定肽序列,分析出1个肽段,氨基酸序列为Thr-Ser-Gly-Met-Pro,相对分子质量为491.56。
In order to investigate the structure of peptides fromiron-chelating black-bone silky fowloligopeptides(BSFP-Fe),BSFP-Fe wasprepared by using black-bone silky fowl as raw material andiron chloride as iron resource. The total protein content,chelating rate and yield were determined.Results showed that the total protein content,chelating rate and yield were 56.79 ±0.18%,83.53 ±0.15% and 40.86±0.14%,respectively. Then,BSFP-Fe was separated and purified by reversed phase high performance liquid chromatography(RP-HPLC).A total of4 fractions were collected and subjected to Q-TOF mass spectrometer to identify peptide sequences. One peptide,Thr-Ser-Gly-MetPro,was identified and its molecular weight was 491.56 u. This studyconfirmed the structure of peptides from BSFP-Feand provided certain theoretical support for the development of BSFP-Fe as iron supplement food.
In order to investigate the structure of peptides fromiron-chelating black-bone silky fowloligopeptides(BSFP-Fe),BSFP-Fe wasprepared by using black-bone silky fowl as raw material andiron chloride as iron resource. The total protein content,chelating rate and yield were determined.Results showed that the total protein content,chelating rate and yield were 56.79 ±0.18%,83.53 ±0.15% and 40.86±0.14%,respectively. Then,BSFP-Fe was separated and purified by reversed phase high performance liquid chromatography(RP-HPLC).A total of4 fractions were collected and subjected to Q-TOF mass spectrometer to identify peptide sequences. One peptide,Thr-Ser-Gly-MetPro,was identified and its molecular weight was 491.56 u. This studyconfirmed the structure of peptides from BSFP-Feand provided certain theoretical support for the development of BSFP-Fe as iron supplement food.
引文
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[21]慕现敏.鱼鳞钙离子结合活性肽的分离纯化及其生理活性研究[D].青岛:中国海洋大学,2013.
[22]NIU Rui,YU Jiansheng.Antioxidant activity and purification of pollockpeptides[J].Journal of Food Science and Biotechnology,2010,29(4):562-566.(in Chinese)
[23]ZHANG Q X,WU H,LING Y F.Isolation and identification of antioxidant peptides derived from whey protein enzymatic hydrolysate by consecutive chromatography and Q-TOF MS[J].J Dairy Res,2013,80(3):367-373.
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[26]TORRES-FUENTES C,ALAIZ M,VIOQUE J.Iron-chelating activity of chickpea protein hydrolysate peptides[J].Food Chem,2012,134:1585-1588.
[2]TIAN Y G,XIE M Y,WANG W Y,et al.Determination ofcarnosine in Black-Bone Silky Fowl(Gallus gallusdomesticus Brisson)and commonchicken by HPLC[J].Eur Food Res Technol,2007,226:311-314.
[3]LIN Lin,TIAN Yinggang,XIE Mingyong,et al.Research on antioxidant activity in vitro and molecular weight distribution of Black-bone silky fowl bioactive peptides[J].Food Science,2007,28(10):41-45.(in Chinese)
[4]TIAN Yinggang,XIE Mingyong,WU Hongjing,et al.Effects of hexane extract from Black-bone silky fowl on hematopoiesis[J].Pharmacology and Clinics of Chinese Materia Medica,2007,23(1):48-50.(in Chinese)
[5]SUN Ning,WANG Zhigeng,MEI Lin,et al.Experiment on the effects of hydrolysedcasein bound iron in ironsupplementation on rats[J].Journal of Food Science and Biotechnology,2013,32(3):330-335.(in Chinese)
[6]LIU Yong,YE Xian,WEI Shoulian.Optimization of the preparation process for iron chelate of Tilapia scale collagen peptide[J].Fine Chemicals,2013,30(7):761-765.(in Chinese)
[7]ZHENG Jiong,WANG Xuerong,KAN Jianquan.Study on preparation of haemoglobin polypeptide-ironanditsanti-anaemiafunction[J].Science and Technology of Food Industry,2009,30(10):312-314.(in Chinese)
[8]LV Y,LIU Q,BAO X,et al.Identification and characteristics of ironchelatingpeptides from soybean protein hydrolysates using IMAC-Fe3+[J].J Agr Food Chem,2009,57(11):4593-4597.
[9]TORRES-FUENTES C,ALAIZ M,VIOQUE J.Iron-chelating activity ofchickpea protein hydrolysate peptides[J].Food Chem,2012,134(3):1585-1588.
[10]HUANG G,REN Z,JIANG J.Separation of iron-binding peptides fromshrimp processing by-products hydrolysates[J].Food Bioprocess Tech,2011,4(8):1527-1532.
[11]SUN Ning,WANG Zhigeng,MEI Lin,et al.Experiment on the effects of hydrol Ysed casein bound iron in iron supplementation on rats[J].Journal of Food Science and Biotechnology,2013,32(3):330-335.(in Chinese)
[12]LIU Wenying,LU Jun,GU Ruizeng,et al.Stability of iron-chelating Bback-bone silky fowl oligopeptides[J].Food and Fermentation Industries,2014,40(10):124-128.(in Chinese)
[13]SONG Shasha,GAO Fei,REN Difeng,et al.Preparation and infrared spectrum identification of black-bonesilky fowl oligopeptide-iron(Ⅱ)chelates[J].Food and Fermentation Industries,2013,39(6):13-17.(in Chinese)
[14]LIU Wenying,GU Ruizeng,LU Jun,et al.Preparation and stability of black-bone silky fowl oligopeptides[J].Food Science and Technology,2014,39(12):157-160.(in Chinese)
[15]LIU Wenying,MA Rongqing,JIN Zhentao,et al.Antioxidant activities in vitro of black-bone silky fowl oligopeptides[J].Food and Fermentation Industries,2010,36(10):19-23.(in Chinese)
[16]国家卫生部.食品中蛋白质的测定:GB/T5009.5-2003[S].北京:中国标准出版社,2003.
[17]LIU Wenying,GU Ruizeng,LIN Feng,et al.Study on separation and purification of marine collagen oligopeptidesand their antioxidant properties[J].Science and Technology of Food Industry,2012,33(18):131-134.(in Chinese)
[18]GU R Z,LIU W Y,LIN F,et al.Antioxidant and angiotensin I-converting enzyme inhibitory properties of oligopeptides derived from black-bone silky fowl(Gallus gallusdomesticus Brisson)muscle[J].Food Res Int,2012,49(1):326-333.
[19]GU R Z,LI C Y,LIU W Y,et al.Angiotensin I-converting enzyme inhibitory activity of low-molecular-weight peptides from Atlantic salmon(Salmo salar L.)skin[J].Food Res Int,2011,44(5):1536-1540.
[20]LIU W Y,LU J,GAO F,et al.Preparation,characterization and identification of calcium-chelating Atlantic salmon(Salmo salar L.)ossein oligopeptides[J].Eur Food Res Technol,2015,DOI:10.1007/s00217-015-2510-2
[21]慕现敏.鱼鳞钙离子结合活性肽的分离纯化及其生理活性研究[D].青岛:中国海洋大学,2013.
[22]NIU Rui,YU Jiansheng.Antioxidant activity and purification of pollockpeptides[J].Journal of Food Science and Biotechnology,2010,29(4):562-566.(in Chinese)
[23]ZHANG Q X,WU H,LING Y F.Isolation and identification of antioxidant peptides derived from whey protein enzymatic hydrolysate by consecutive chromatography and Q-TOF MS[J].J Dairy Res,2013,80(3):367-373.
[24]CHALKLEY R J,BURLINGAME A L.Identification of Glc NAcylation sites of peptides and alpha-crystallin using Q-TOF mass spectrometry[J].J Am Soc Mass Spectrom,2001,12(10):1106-1113.
[25]林慧敏.带鱼下脚料酶解小肽亚铁螯合物结构鉴定及其生物活性研究[D].福州:福建农林大学,2012.
[26]TORRES-FUENTES C,ALAIZ M,VIOQUE J.Iron-chelating activity of chickpea protein hydrolysate peptides[J].Food Chem,2012,134:1585-1588.