Purification and comparison of enzymatic properties of endogenous transglutaminase between silver carp and black carp

Authors

YI Lin, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China;National R & D Branch Center for Conventional Freshwater Fish Processing 〔Wuhan〕, Huazhong Agricultural University, Wuhan, Hubei 430070, China
AN Yueqi, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China;National R & D Branch Center for Conventional Freshwater Fish Processing 〔Wuhan〕, Huazhong Agricultural University, Wuhan, Hubei 430070, China
LIU Ru, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China;National R & D Branch Center for Conventional Freshwater Fish Processing 〔Wuhan〕, Huazhong Agricultural University, Wuhan, Hubei 430070, China
HU Yang, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China;National R & D Branch Center for Conventional Freshwater Fish Processing 〔Wuhan〕, Huazhong Agricultural University, Wuhan, Hubei 430070, China
XIONG Shanbai, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China;National R & D Branch Center for Conventional Freshwater Fish Processing 〔Wuhan〕, Huazhong Agricultural University, Wuhan, Hubei 430070, ChinaFollow

Corresponding Author(s)

熊善柏（1963—），男，华中农业大学教授，硕士。E-mail: xiongsb@mail.hzau.edu.cn

Abstract

Objective: This study aimed to investigate the differences in enzymatic properties of endogenous transglutaminase （TGase） in silver carp and black carp. Methods: STG and BTG were purified from the muscle of silver carp and black carp, respectively, by 80% ammonium sulfate precipitation, Q-Sepharose FF, and Sephacryl S-200 HR chromatographies. Two enzymes were analyzed for relative molecular weights, peptide sequences, secondary structures, optimal reaction conditions, and thermal inactivation kinetics. Results: The purified STG and BTG showed similar relative molecular weights, of which the enzyme activities were 14.34 U/mg and 12.67 U/mg, respectively. Both enzymes showed differences in peptide sequences. The secondary structures of them were mainly the β-fold, though the content of β-fold in STG was slightly higher than that of BTG. The optimal temperatures for STG and BTG were both 50 ℃, and the optimal pH values were 8.0 and 7.5, respectively. The enzymes required Ca²⁺ up to 1 mmol/L for full activation. The activities of STG and BTG were enhanced by DTT, whereas PMSF, NH₄Cl, NEM, EDTA, Cu²⁺, Ba²⁺, Zn²⁺, and Mg²⁺ showed inhibitory effects. When the temperature was 37~50 ℃, the passivations of STG and BTG by thermal treatment conformed to the first-order exponential decay kinetics with similar values of E_a. Conclusion: The primary and secondary structures of STG and BTG exhibited obvious differences, yet they still exhibited similar properties in terms of optimal reaction conditions and thermal inactivation kinetics.

Publication Date

12-26-2023

First Page

4

Last Page

12

DOI

10.13652/j.spjx.1003.5788.2023.80841

Recommended Citation

Lin, YI; Yueqi, AN; Ru, LIU; Yang, HU; and Shanbai, XIONG (2023) "Purification and comparison of enzymatic properties of endogenous transglutaminase between silver carp and black carp," Food and Machinery: Vol. 39: Iss. 10, Article 1.
DOI: 10.13652/j.spjx.1003.5788.2023.80841
Available at: https://www.ifoodmm.cn/journal/vol39/iss10/1

References

[1] WORRATAO A, YONGSAWATDIGUL J. Purification and characterization of transglutaminase from tropical tilapia （Oreochromis niloticus）[J]. Food Chemistry, 2005, 93（4）: 651-658.
[2] 沈晓蕾, 李向红, 俞健, 等. 大豆分离蛋白、木薯淀粉与转谷氨酰胺酶组合对鲢鱼鱼糜凝胶品质的影响[J]. 食品与机械, 2019, 35（9）: 26-31. SHEN X L, LI X H, YU J, et al. Effect of combination of soy protein isolate, tapioca starch and transglutamin-ase on gel quality of carp surimi[J]. Food & Machinery, 2019, 35（9）: 26-31.
[3] 贾丹, 刘茹, 刘明菲, 等. 转谷氨酰胺酶对鳙鱼糜热诱导胶凝特性的影响[J]. 食品科学, 2013, 34（9）: 37-41. JIA D, LIU R, LIU M F, et al. Effect of transglutaminase on heat-induced gel properties of bighead carp （Aristichthys nobilis） surimi[J]. Food Science, 2013, 34（9）: 37-41.
[4] 张梦玲, 张晋, 熊善柏, 等. 转谷氨酰胺酶及其在鱼糜制品加工中的应用[J]. 食品研究与开发, 2016, 37（24）: 190-195. ZHANG M L, ZHANG J, XIONG S B, et al. Transglutaminase and its application in processing of surimi product[J]. Food Research and Development, 2016, 37（24）: 190-195.
[5] 何立, 李细毛, 胡冰悦, 等. 3种鱼糜蛋白质特性及油炸特性比较[J]. 食品与机械, 2022, 38（6）: 40-44. HE L, LI X M, HU B Y, et al. Comparative study on protein and frying characteristics of three surimi[J]. Food & Machinery, 2022, 38（6）: 40-44.
[6] 贾丹. 青鱼肌肉蛋白质及其凝胶特性的研究[D]. 武汉: 华中农业大学, 2016: 36. JIA D. Study on protein and gelation properties of surimi from black carp[D]. Wuhan: Huazhong Agricultural University, 2016: 36.
[7] ZHANG Y, SIMPSON B K. Food-related transglutaminase obtained from fish/shellfish[J]. Critical Reviews in Food Science and Nutrition, 2020, 60（19）: 3 214-3 232.
[8] 李金玲, 叶蕾蕾, 尤娟, 等. 鲢内源性转谷氨酰胺酶的纯化及其性质[J]. 华中农业大学学报, 2018, 37（6）: 105-112. LI J L, YE L L, YOU J, et al. Purification and some enzymatic properties of transglutaminase from silver carp[J]. Journal of Huazhong Agricultural, 2018, 37（6）: 105-112.
[9] 孙静静, 罗自生, 吴翔, 等. 草鱼中内源性转谷氨酰胺酶特性的研究[J]. 中国食品学报, 2012, 12（9）: 67-72. SUN J J, LUO Z S, WU X, et al. Study on the characterization of transglutaminase from grass carp[J]. Journal of Chinese Institute of Food Science and Technology, 2012, 12（9）: 67-72.
[10] 娄忠纬, 徐坤华, 王潇, 等. 鳙内源性转谷氨酰胺酶特性的研究[J]. 南方水产科学, 2014, 10（6）: 72-77. LOU Z W, XU K H, WANG X, et al. Characteristics of transglutaminase from bighead carp[J]. South China Fisheries Science, 2014, 10（6）: 72-77.
[11] HEMUNG B O, YONGSAWATDIGUL J. Partial purification and characterization of transglutaminase from threadfin bream（Nemipterus sp.） liver[J]. Journal of Food Biochemistry, 2008, 32（2）: 182-200.
[12] LOWRY O H, ROSEBROUGH N J, FARR A L, et al. Protein measurement with the folin phenol reagent[J]. Journal of Biological Chemistry, 1951, 193（1）: 265-275.
[13] 王耀冉, 赵妍, 陈明杰, 等. 8种食用菌蛋白及其酶解产物抗氧化活性研究[J]. 食品与机械, 2022, 38（9）: 134-138, 152. WANG Y R, ZHAO Y, CHEN M J, et al. Study on antioxidant activities of proteins and enzymatic hydrolysates of eight edible fungi[J]. Food & Machinery, 2022, 38（9）: 134-138, 152.
[14] ALHASANI H A W, AL-YOUNIS Z K. Extraction, purification and characterization of transglutaminase from some plants[J]. IOP Conference Series: Earth and Environmental Science, 2021, 910: 012061.
[15] NOZAWA H, CHO S Y, SEKI N. Purification and characterization of transglutaminase from squid gill[J]. Fisheries Science, 2001, 67（5）: 912-919.
[16] SIRIKHARIN R, SDERHLL I, SDERHLL K. Characterization of a cold-active transglutaminase from a crayfish, Pacifastacus leniusculus[J]. Fish & Shellfish Immunology, 2018, 80: 546-549.
[17] LAKSONO U T, SUPRIHATIN, NURHAYATI T, et al. Purification of endogenous transglutaminase from daggertooth pike conger fish （Muraenesox cinerus） meat[J]. International Food Research Journal, 2021, 28（5）: 1 030-1 037.
[18] YASUEDA H, KUMAZAWA Y, MOTOKI M. Purification and characterization of a tissue-type transglutaminase from red sea bream （Pagrus major）[J]. Bioscience, Biotechnology, and Biochemistry, 1994, 58（11）: 2 041-2 045.
[19] NOGUCHI K, ISHIKAWA K, YOKOYAMA K, et al. Crystal structure of red sea bream transglutaminase[J]. Journal of Biological Chemistry, 2001, 276（15）: 12 055-12 059.
[20] 崔文璟, 杜坤, 周丽, 等. 吸水链霉菌谷氨酰胺转氨酶两种同源异形体的鉴定及性质分析[J]. 中国生物制品学杂志, 2013, 26（7）: 949-953. CUI W J, DU K, ZHOU L, et al. Identification and properties of two isoforms of Streptomyces hygroscopicus transglutaminase[J]. Chinese Journal of Biologicals, 2013, 26（7）: 949-953.
[21] SIDAURUK W S, TATI N, SUPTIJAH P, et al. Characterization of endogenous transglutaminase enzyme of yellow pike conger's liver[J]. Jurnal Pengolahan Hasil Perikanan Indonesia, 2017, 20: 582.
[22] ZHANG Y, HE S, SIMPSON B K. A cold active transglutaminase from antarctic krill （Euphausia superba）: Purification, characterization and application in the modification of cold-set gelatin gel[J]. Food Chemistry, 2017, 232: 155-162.
[23] DE SOUZA C F V, FACCIN D J L, MERTINS O, et al. Kinetics of thermal inactivation of transglutaminase from a newly isolated bacillus circulans BL32[J]. Journal of Chemical Technology & Biotechnology, 2009, 84（10）: 1 567-1 575.
[24] ZHANG Y, LI C, GEARY T, et al. Contribution of special structural features to high thermal stability of a cold-active transglutaminase[J]. Journal of Agricultural and Food Chemistry, 2020, 68（30）: 7 935-7 945.
[25] BINSI P K, SHAMASUNDAR B A. Purification and characterisation of transglutaminase from four fish species: Effect of added transglutaminase on the viscoelastic behaviour of fish mince[J]. Food Chemistry, 2012, 132（4）: 1 922-1 929.
[26] WANG B, WANG A, CAO Z, et al. Characterization of a novel highly thermostable esterase from the gram-positive soil bacterium streptomyces lividans TK64[J]. Biotechnology and Applied Biochemistry, 2016, 63（3）: 334-343.
[27] CUI L, DU G, ZHANG D, et al. Purification and characterization of transglutaminase from a newly isolated Streptomyces hygroscopicus[J]. Food Chemistry, 2007, 105: 612-618.
[28] QUEIRS R P, GOUVEIA S, SARAIVA J A, et al. Impact of pH on the high-pressure inactivation of microbial transglutaminase[J]. Food Research International, 2019, 115: 73-82.