Proteomic Analysis of Xuanwei Ham by Two-Dimensional Gel Electrophoresis

Abstract

Abstract: In the present study, a method for the proteomic analysis of Xuanwei ham was established using two-dimensional gel electrophoresis (2-DE). Muscle protein from Xuanwei ham was extracted with trichloroacetic acid-acetone, quantified, and analyzed by isoelectric focusing electrophoresis and SDS-PAGE. After electrophoresis, the gels were stained with coomassie brilliant blue, and the observed protein spots were analyzed using imageMaster 2D platinum software and one of them was selected and analyzed by matrix-assisted laser desorption ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF/TOF MS) after tripsin digestion. The obtained peptide mapping fingerprint was analyzed using Mascot software. A total of 1749 clear protein spots were observed in the 2-DE pattern of Xuanwei ham, which were mainly distributed over a pH range of 4.0－7.0. The tested protein spot was identified as gamma-2-actin protein and found to have a Mascot score of 129 (threshold was 67). These results demonstrate the successful establishment of a 2-DE method for the proteomic analysis of Xuanwei ham.

Key words: Xuanwei ham, proteomics, two-dimensional electrophoresis

CLC Number:

TS207.3

References

[1]乔发东.宣威火腿标准化生产与品质改良技术研究[D][J].北京中国农业大学,2004,6:48-58
[2]LARREA V, HERNANDO I, QUILES A, et al. .Changes in proteins during Teruel dry-cured ham processing [J].,2006,74:586-
[3]TOLDRá F, RICO E, FLORES J.Cathepsin B, D, H and L activity in the Processing of dry-cured ham[J].Journal of the Science of Food and Agriculture,1993,62(4):157-161
[4]TOLDRá F, FLORES M, SANZ Y.Dry cured ham falvour: emzymatic generation and process influence[J].Food Chemistry,1997,59(4):523-530
[5]LARREA V, HERNANDO I, QUILES A, et al.Changes in proteins during Teruel dry-cured ham processing[J].Meat Science,2006,74:586-593
[6]FLORES M, BARAT J M, ARISTOY M C, et al.Accelerated processing of dry-cured hamPart 2. In?uence of brine thawing/salting operation on proteolysis and sensory acceptability[J].. Meat Science,2006,72:766-772
[7]DI LUCIA A, PICARIELLA G, CACACE G, et al.Proteomic analysis of water soluble and myofibrillar protein changes occuring in dry cured ham[J].Meat Science,2005,69:479-491
[8]MORA L, SENTANDREU M A, TOLDRá F.Identification of small troponin T peptides generated in dry-cured ham[J].Food Chemistry,2010,123:691-697
[9]BRADFORD M M.A rapid and sensitive method for the quantitation of microgram of protein utilizing the principle of protein dye binding[J].Analytical Biochemistry,1976,72:248-254
[10]THéRON L, SAYD T, PINGUET J, et al.Proteomic analysis of semimembranosus and biceps femoris muscles from Bayonne dry-cured ham[J].Meat Science,2011,88(1):82-90
[11]BODZON-KULAKOWSKA A, BIERCZYNSKA-KRZYSIK A, DYLAG T, et al.Methods for samples preparation in proteomic research[J].Journal of Chromatography B,2007,849:1-31
[12]HOLLUNG K, VEISETH E, JIA X H, et al.Application of proteomics to understand the molecular mechanisms behind meat quality[J].Meat Science,2007,77:97-104
[13]KVASNICKA F.Proteomics: general strategies and application to nutritionally relevant proteins[J].Journal of Chromatography B,2003,787:77-89
[14]CARBONARO M.Proteomics: present and future in food quality evaluation[J].Trends in Food Science & Technology,2004,15:209-216
[15]HAN Jianzhong, Wang Yanbo.Proteomics: present and future in food science and technology[J].Trends in Food Science & Technology,2008,19:26-30