|本期目录/Table of Contents|

 LIN Xinyu,ZHANG Ying,ZHANG Weini,et al.Inhibitory kinetics of β-N-acetyl-D-glucosaminidase from Nile tilapia (Oreochromis niloticus) by cadmium[J].Chinese Journal of Applied & Environmental Biology,2018,24(03):609-614.[doi:10.19675/j.cnki.1006-687x.2017.08027]





Inhibitory kinetics of β-N-acetyl-D-glucosaminidase from Nile tilapia (Oreochromis niloticus) by cadmium
1福建省兽医中药与动物保健重点实验室(福建农林大学)? 福州 350002 2宁德市动物疾病预防控制中心 宁德 352100
LIN Xinyu1 ZHANG Ying1 ZHANG Weini1 WU Fangda2 HUANG Yifang1 & HUANG Xiaohong1**
1 Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health, Fujian Agriculture and Forestry University, Fuzhou 350002, China 2 Ningde Centers for Animal Disease Control and Prevention, Ningde 352100, China
inhibitory kinetics β-N-acetyl-D-glucosaminidase Nile tilapia Cd2+
Q556.2 : X174
镉离子污染对水生生物的繁殖造成了不利影响,N-乙酰-β-D-氨基葡萄糖苷酶(NAGase)被认为参与了动物的受精过程. 为了解镉离子对罗非鱼繁殖的影响,以尼罗罗非鱼精巢分离获得的NAGase酶为研究对象,采用底物反应动力学方法,建立镉离子对NAGase酶的失活模型,探究镉离子对该酶活性的影响. 结果显示,镉离子对NAGase表现出了可逆的抑制,对NAGase抑制的IC50为40.95 mmol/L.镉离子对NAGase的抑制机理为竞争型抑制,抑制常数KI为17.13 mmol/L,同时测定了微观失活速度常数. 本研究表明镉离子对NAGase的抑制类型为竞争型的可逆抑制,结果可为后期研究镉离子对罗非鱼繁殖性能的研究提供理论基础.(图8 表1 参33)
Cadmium (Cd2+) pollution in an aquatic environment can negatively affect certain reproductive parameters of aquatic animals. β-N-acetyl-D-glucosaminidase (NAGase) is considered to play an important role in the fertilization process. The aim of the present study was to investigate the effect of Cd2+ on the activity of NAGase purified from the testis of Nile tilapia, toward contributing new knowledge on the breadth of negative effects of Cd2+ for Nile tilapia production. The kinetic method of substrate reaction was used for this assessment, and an inhibitory model was established to study the kinetics of NAGase under inhibition by Cd2+. The results showed that Cd2+ could reversibly inhibit the enzymatic activity of NAGase, and the half-maximal inhibitory concentration was estimated to be 40.95 mmol/L. Cd2+ was found to be a competitive inhibitor of NAGase, and the inhibitory constant was determined to be 17.13 mmol/L. The microscopic rate constants of inactivation were also determined. Together, these findings demonstrate that Cd2+ is a reversible inhibitor that can competitively inhibit NAGase. These results may provide a theoretical foundation for further studies on the reproduction of tilapia.


1 Rao Cv Fau - Edgerton LA, Edgerton LA. Dissimilarity of corpora lutea within the same ovaries or those from right and left ovaries of pigs during the oestrous cycle [J]. J Reprod Fertil, 1984, 70 (1): 61-66 2 Zhang WN, Bai DP, Huang YF, Hu CW, Chen QX,Huang XH. Enzymatic characterizations and activity regulations of N-acetyl-beta-D-glucosaminidase from the spermary of Nile tilapia (Oreochromis niloticus) [J]. J Biosci Bioeng, 2014, 117 (2): 153-157 3 Koyanagi R,Honegger TG. Molecular cloning and sequence analysis of an ascidian egg beta-N-acetylhexosaminidase with a potential role in fertilization [J]. Dev Growth Differ, 2003, 45 (3): 209-218 4 Sarosiek B, Glogowski J, Cejko BI, Kujawa R, Szczepkowski M, Kuzminski H, Dobosz S,Kowalski RK. Inhibition of beta-N-acetylglucosaminidase by acetamide affects sperm motility and fertilization success of rainbow trout (Oncorhynchus mykiss) and Siberian sturgeon (Acipenser baerii) [J]. Theriogenology, 2014, 81 (5): 723-732 5 Cattaneo F, Ogiso M, Hoshi M, Perotti ME,Pasini ME. Purification and characterization of the plasma membrane glycosidases of Drosophila melanogaster spermatozoa [J]. Insect Biochem Mol Biol, 2002, 32 (8): 929-941 6 Miller DJ, Gong X,Shur BD. Sperm require beta-N-acetylglucosaminidase to penetrate through the egg zona pellucida [J]. Development, 1993, 118 (4): 1279-1289 7 Martinez ML, Martelotto L,Cabada MO. Purification and biological characterization of N-acetyl beta-D glucosaminidase from Bufo arenarum spermatozoa [J]. Mol Reprod Dev, 2000, 57 (2): 194-203 8 傅鹏, 王飞, 马秀平, 王兰. 沁河沉积物重金属垂直分布特征与风险评价[J]. 应用与环境生物学报, 2013, 19 (2): 305-312 [Fu P, Wang F, Ma XP, Wang L. Vertical distribution of heavy metals in sediments of Qinhe river and its risk assessment [J]. Chin J Appl Environ Boil, 2013, 19 (2): 305-312] 9 Chahid A, Hilali M, Benlhachimi A,Bouzid T. Contents of cadmium, mercury and lead in fish from the Atlantic sea (Morocco) determined by atomic absorption spectrometry [J]. Food Chem, 2014, 147: 357-360 10 Taweel A, Shuhaimi-Othman M, Ahmad AK. Assessment of heavy metals in tilapia fish (Oreochromis niloticus) from the Langat River and Engineering Lake in Bangi, Malaysia, and evaluation of the health risk from tilapia consumption [J]. Ecotoxicol Environ Saf, 2013, 93: 45-51 11 Zhou Q, Zhang J, Fu J, Shi J, Jiang G. Biomonitoring: an appealing tool for assessment of metal pollution in the aquatic ecosystem [J]. Anal Chim Acta, 2008, 606 (2): 135-150 12 Tang W, Shan B, Zhang W, Zhang H, Wang L,Ding Y. Heavy metal pollution characteristics of surface sediments in different aquatic ecosystems in eastern China: a comprehensive understanding [J]. PLoS ONE, 2014, 9 (9): e108996 13 Luo Y, Shan D, Zhong H, Zhou Y, Chen W, Cao J, Guo Z, Xiao J, He F, Huang Y, Li J, Huang H, Xu P. Subchronic effects of cadmium on the gonads, expressions of steroid hormones and sex-related genes in tilapia Oreochromis niloticus [J]. Ecotoxicology, 2015, 24 (10): 2213-2223 14 Takiguchi M,Yoshihara S. New aspects of cadmium as endocrine disruptor [J]. Environ Sci, 2006, 13 (2): 107-116 15 Garcia-Santos S, Fontainhas-Fernandes A, Monteiro SM, Wilson JM. Effects of exposure to cadmium on some endocrine parameters in tilapia, Oreochromis niloticus [J]. Bull Environ Contam Toxicol, 2013, 90 (1): 55-59 16 Vergilio CS, Moreira RV, Carvalho CE, Melo EJ. Evolution of cadmium effects in the testis and sperm of the tropical fish Gymnotus carapo [J]. Tissue Cell, 2015, 47 (2): 132-139 17 Xu LC, Sun H, Wang SY, Song L, Chang HC,Wang XR. The roles of metallothionein on cadmium-induced testes damages in Sprague-Dawley rats [J]. Environ Toxicol Pharmacol, 2005, 20 (1): 83-87 18 Oliveira H, Spano M, Santos C,Pereira Mde L. Adverse effects of cadmium exposure on mouse sperm [J]. Reprod Toxicol, 2009, 28 (4): 550-555 19 de Oliveira MM, Ribeiro T, Orlando TM, de Oliveira DG, Drumond MM, de Freitas RT,Rosa PV. Effects crude protein levels on female Nile tilapia (Oreochromis niloticus) reproductive performance parameters [J]. Anim Reprod Sci, 2014, 150 (1-2): 62-69 20 Wang R, Feng XB, Wang WX. In vivo mercury methylation and demethylation in freshwater tilapia quantified by mercury stable isotopes [J]. Environ Sci Technol, 2013, 47(14): 7949-7957 21 Wang WX. Incorporating exposure into aquatic toxicological studies: an imperative [J]. Aquat Toxicol, 2011, 105 (3-4 Suppl): 9-15 22 Wang R, Wang WX. Contrasting mercury accumulation patterns in tilapia (Oreochromis niloticus) and implications on somatic growth dilution [J]. Aquat Toxicol, 2012, 114-115: 23-30 23 沈林松, 张颖, 张伟妮, 赖育河,黄小红. 果糖对莆田黑猪精液中NAGase的抑制动力学[J]. 应用与环境生物学报, 2015, 21 (1): 46-50 [Shen LS, Zhang Y, Zhang WN, Lai YH, Huang XH. Inhibitory kinetics of fructose on NAGase in semen of Putian black pigs [J]. Chin J Appl Environ Boil, 2015, 21 (1): 46-50] 24 张伟妮, 陈晓佳, 沈林松, 黄一帆,黄小红. 强氯精对尼罗罗非鱼精巢N-乙酰-β-D-氨基葡萄糖苷酶的失活动力学[J]. 应用与环境生物学报, 2017, 23 (2): 306-311 [Zhang WN, Chen XJ, Shen LS, Huang YF, Huang XH. Inactivation kinetics of N-Acetyl-β-D-glucosaminidase from the spermary of Nile tilapia (Oreochromis niloticus) by trichloroisocyanuric acid [J]. Chin J Appl Environ boil, 2017, 23 (2): 306-311] 25 Lowry OH, Rosebrough NJ, Farr AL,Randall RJ. Protein measurement with the Folin phenol reagent [J]. J Biol Chem, 1951, 193 (1): 265-275 26 Tsou CL. Kinetics of substrate reaction during irreversible modification of enzyme activity [J]. Adv Enzymol Relat Areas Mol Biol,?1988, 61: 381-436 27 Zhang JP, Hu YH, Wang Q, Wang W, Wang Y, Yan JH,Chen QX. Inhibitory kinetics of beta-N-acetyl-D-glucosaminidase from green crab (Scylla serrata) by zinc ion [J]. J Agric Food Chem, 2010, 58 (15): 8763-8767 28 Luo J, Pei S, Jing W, Zou E,Wang L. Cadmium inhibits molting of the freshwater crab Sinopotamon henanense by reducing the hemolymph ecdysteroid content and the activities of chitinase and N-acetyl-beta-glucosaminidase in the epidermis [J]. Comp Biochem Physiol C Toxicol Pharmacol, 2015, 169: 1-6 29 Leoni G, Bogliolo L Fau - Deiana G, Deiana G Fau - Berlinguer F, Berlinguer F Fau - Rosati I, Rosati I Fau - Pintus PP, Pintus Pp Fau - Ledda S, Ledda S Fau - Naitana S,Naitana S. Influence of cadmium exposure on in vitro ovine gamete dysfunction [J]. Reprod Toxicol,?2002, 16 (4): 371-377 30 Xie XL, Huang QS, Gong M, Du J, Yang Y,Chen QX. Inhibitory kinetics of beta-N-acetyl-D-glucosaminidase from prawn (Litopenaeus vannamei) by zinc ion [J]. IUBMB Life, 2009, 61 (2): 163-170 31 Xie JJ, Chen QX, Zhang JP, Wang Q,Yang XM. Inhibitory kinetics of mercuric ion on the activity of beta-N-acetyl-D-glucosaminidase from green crab (Scylla serrata) [J]. Int J Biol Macromol, 2006, 39 (4-5): 159-164 32 Zhang WN, Chen QX, Lin XY, Huang XH, Huang YF. Irreversible inhibitory kinetics of mercuric ion on N-acetyl-beta-D-glucosaminidase from Nile tilapia (Oreochromis niloticus) [J]. Aquat Toxicol, 2014, 154: 163-167 33 Pan C, Liu HD, Gong Z, Yu X, Hou XB, Xie DD, Zhu XB, Li HW, Tang JY, Xu YF, Yu JQ, Zhang LY, Fang H, Xiao KH, Chen YG, Wang JY, Pang Q, Chen W, Sun JP. Cadmium is a potent inhibitor of PPM phosphatases and targets the M1 binding site [J]. Sci Rep, 2013, 3: 2333


 HUANG Xiaohong,CHEN Honghui,JIN Yandong,et al.Effects of Amino Acids on N-acetyl-β-D-glucosaminidase Activity of Eriocheir sinensis[J].Chinese Journal of Applied & Environmental Biology,2009,15(03):207.[doi:10.3724/SP.J.1145.2009.00207]
[2]张伟妮,陈欣颖,黄小红,等.4种重金属离子对克氏原螯虾(Procambarus clarkii)NAGase活力的影响[J].应用与环境生物学报,2012,18(06):943.[doi:10.3724/SP.J.1145.2012.00943]
 ZHANG Weini,CHEN Xinying,HUANG Xiaohong,et al.Effects of Four Heavy Metal Ions on NAGase of Procambarus clarkii[J].Chinese Journal of Applied & Environmental Biology,2012,18(03):943.[doi:10.3724/SP.J.1145.2012.00943]
[3]沈林松,张 颖,张伟妮,等.果糖对莆田黑猪精液中NAGase的抑制动力学[J].应用与环境生物学报,2015,21(01):46.[doi:10.3724/SP.J.1145.2014.07013]
 SHEN Linsong,ZHANG Ying,ZHANG Weini,et al.Inhibitory kinetics of fructose on NAGase in semen of Putian black pigs[J].Chinese Journal of Applied & Environmental Biology,2015,21(03):46.[doi:10.3724/SP.J.1145.2014.07013]
 ZHANG Weini,CHEN Xiaojia,SHEN Linsong,et al.Inactivation kinetics of N-Acetyl-β-D-glucosaminidase from the spermary of Nile tilapia (Oreochromis niloticus) by Trichloroisocyanuric acid[J].Chinese Journal of Applied & Environmental Biology,2017,23(03):306.[doi:10.3724/SP.J.1145.2016.04033]
[5]林心宇 张颖 张伟妮 吴方达 黄一帆 黄小红 **.镉离子对尼罗罗非鱼精巢N-乙酰-β-D-氨基葡萄糖苷酶的失活动力学研究[J].应用与环境生物学报,2018,24(04):1.[doi:10.3724/SP.J.1145.2017.08027]
 LIN Xinyu,ZHANG Ying,ZHANG Weini,et al.Inhibitory kinetics of β-N-acetyl-D-glucosaminidase from Nile tilapia (Oreochromis niloticus) by cadmium[J].Chinese Journal of Applied & Environmental Biology,2018,24(03):1.[doi:10.3724/SP.J.1145.2017.08027]

更新日期/Last Update: 2018-06-30