Parvalbumin (PV) is the main allergen of fish. To understand parvalbumin isotypes, amino acid sequence and protein modification in fish, silver carp muscle was used as research object, PV isotypes were isolated and purified by salting out. High-performance liquid chromatography combined with size exclusion chromatography and high-resolution mass spectrometry method were used in this study, and their amino acid sequence and protein modification were explored. The results showed that PV made from silver carp muscle contained three isotypes (I, II and III), and their average molecular weights were 12.029 0 kDa, 11.581 0 kDa and 11.622 0 kDa, respectively. The amino acid sequences of PVI, PVII and PVIII) matched 74%, 100% and 99% respectively with the amino acid sequences of PV isotypes Q804W2, Q6IMW7 and Q9I8V0 in the protein database, and PVII and PVIII had 89% and 88% similarities with the reported PV isotypes of silver carp gi 209902357 and gi 209902355, respectively. In protein modification, except PVI, which only had acylation modification, the other two isotypes mainly had phosphorylation, acylation, methylation and ubiquitination modification, and the modification degree of PVII was significantly higher than that of PVIII and PVI. This study provides a new idea for the extraction, purification and identification of different PV isotypes of fish, and provide a scientific basis for the safety evaluation of freshwater fish and their products.
[1] 孙登瑶. 淡水鱼过敏原PV的消减研究及低敏性食品的开发[D].上海:上海师范大学, 2018.
SUN D Y.Reduction of allergens of freshwater fish allergrns and development of low sensitive food[D].Shanghai:Shanghai Normal University, 2018.
[2] LIANG J, TAYLOR S L, BAUMERT J, et al.Effects of thermal treatment on the immunoreactivity and quantification of parvalbumin from Southern hemisphere fish species with two anti-parvalbumin antibodies[J/OL].Food Control, 2020.DOI:10.1016/j.foodcont.2020.107 675.
[3] LOPATA A L, JEEBHAY M F.Airborne seafood allergens as a cause of occupational allergy and asthma[J].Current Allergy & Asthma Reports, 2013, 13(3):288-297.
[4] RUETHERS T, RAITH M, SHARP M F, et al.Characterisation of Ras k 1 a novel major allergen in Indian mackerel and identification of parvalbumin as the major fish allergen in 33 Asia-pacific fish species[J].Clinical & Experimental Allergy, 2017, 48(4):452-463.
[5] RUETHERS T, TAKI A C, JOHNSTON E B, et al.Seafood allergy:A comprehensive review of fish and shellfish allergens[J].Molecular Immunology, 2018, 100:28-57.
[6] RUETHERS T,TAKI A C,KARNANEEDI S, et al.Expanding the allergen repertoire of salmon and catfish[J/OL].Allergy, 2020.DOI:10.1111/j.ALL.14574.
[7] ELSATED S M, AAS K.Characterization of a major allergen (Cod) chemical composition and immunological properties[J].International Archives of Allergy and Applied Immunology, 1970, 38(5):536-548.
[8] SWOBODA I, BUGAJSKA-SCHRETTER A, VERDINO P, et al.Recombinant carp parvalbumin, the major cross-reactive fish allergen:A tool for diagnosis and therapy of fish allergy[J].The Journal of Immunology, 2002, 168(9):4 576-4 584.
[9] VANDO T, HORDVIK I, ENDRESEN C, et al.Characterization of parvalbumin, the major allergen in Alaska pollack, and comparison with codfish allergen M[J].Molecular Immunology, 2005, 42(3):345-353.
[10] BUGAJSKA-SCHRETTER A, GROTE M, VANGELISTA L, et al.Purification, biochemical, and immunological characterisation of a major food allergen:Different immunoglobulin E recognition of the apo- and calcium-bound forms of carp parvalbumin[J].Gut, 2000, 46(5):661-669.
[11] CAI Q F, LIU G M, LI T, et al.Purification and characterization of parvalbumins, the major allergens in red stingray (Dasyatis akajei)[J].Journal of Agricultural & Food Chemistry, 2010, 58(24):12 964-12 969.
[12] 农业农村部渔业渔政管理局. 中国渔业统计年鉴[M].北京:中国农业出版社, 2020.
Fishery Administration Bureau of Ministry of Agriculture.China Fishery Statistical Yearbook[M].Beijing:China Agricultural Press, 2020.
[13] 李军, 罗娟, 涂宗财, 等.鲢鱼骨胶原多肽的制备及其抗氧化活性研究[J].食品与发酵工业, 2020, 46(2):222-230.
LI J, LUO J, TU Z C, et al.Study on the preparation and antioxidant activity of collagen polypeptide from silver carp bone[J].Food and Fermentation Industries, 2020, 46(2):222-230.
[14] 孙礼瑞, 林洪, 李振兴, 等.牙鲆(Paralichthys olivaceus)中不同亚型小清蛋白的提纯与鉴定[J].食品科学, 2019, 40(12):308-314.
SUN L D, LIN H, LI Z X, et al.Purification and characterization of parvalbumin isotypes from Japanese flounder (Paralichthys olivaceus)[J].Food Science, 2019, 40(12):308-314.
[15] LIU J, TU Z C, WANG H, et al.Comparative studies on physicochemical properties of bovine serum albuminglucose and galactose conjugates formed by glycation combined with ultrasonic pretreatment[J/OL].International Journal of Food Engineering, 2017.DOI:10.1515/ijfe-2017-0160.
[16] LIU J, TU Z C, ZHANG L, et al.Influence of ultrasonication prior to glycation on the physicochemical properties of bovine serum albumin-galactose conjugates[J].Food Science & Technology Research, 2018, 249(1):35-44.
[17] OFFENGENDEN M, FENTABIL M A, WU J.N-glycosylation of ovomucin from hen egg white[J].Glycoconjugate Journal, 2011, 28(3):113-123.
[18] 蔡秋凤, 曹敏杰, 刘光明, 等.鲢鱼小清蛋白的分离纯化及其单克隆抗体的制备与鉴定[J].中国免疫学杂志, 2011, 27(8):731-734.
CAI Q F, CAO M J, LIU G M, et al.Preparation and identification of monoclonal antibody against silver carp parvalbumin[J].Chinese Journal of Immunology, 2011, 27(8):731-734.
[19] LIU G M, WANG N, CAI Q F, et al.Purification and characterization of parvalbumins from silver carp (Hypophthalmichthy molitrix)[J].Journal of the Science of Food and Agriculture, 2010, 90(6):1 034-1 040.
[20] 李铮. 草鱼主要过敏原小清蛋白亚型纯化鉴定及加工对过敏原影响的研究[D].北京:中国农业大学, 2014.
LI Z.Purification and characterization of parvalbumin isotypes and effect of processing methods on the major allergen of grass carp[D].Beijing:China Agricultural University, 2014.
[21] SUN L R, XU L L, HUANG Y H, et al.Identification and comparison of allergenicity of native and recombinant fish major allergen parvalbumins from Japanese flounder (Paralichthys olivaceus)[J].Food & Function, 2019, 10(10):6 615-6 623..
[22] GUO F F, KUBOTA H, SHIOMI K.Purification, immunological properties and molecular cloning of two allergenic parvalbumins from the crimson sea bream, Evynnis japonica[J].Food Chemistry, 2012, 132(2):835-840.
[23] MARINA P G, JING L, LUDMILLA B, et al.Epitope mapping of Atlantic salmon major allergen by peptide microarray immunoassay[J].International Archives of Allergy & Immunology, 2011, 157(1):31-40.
[24] DIKIC I.Proteasomal and autophagic degradation systems[J].Annual Review of Biochemistry, 2017, 86(1):193-224.
[25] HIRANO H, KIMURA Y, KIMURA A.Biological significance of co- and post-translational modifications of the yeast 26S proteasome[J].Journal of Proteomics, 2016, 134:37-46.
[26] 卢慧, 李衍常, 黄学石.蛋白酶体翻译后修饰功能机制研究[J/OL].中国生物化学与分子生物学报, 2020.http://kns.cnki.net/kcms/detail/11.3870.Q.20201130.1705.002.html.
LU H, LI Y C, HUANG X S.Mechanistic studies of post-translational modifications on the proteasome[J/OL].Chinese Journal of Biochemistry and Molecular Biology, 2020.http://kns.cnki.net/kcms/detail/11.3870.Q.20201130.1705.002.html.
