[1]林建城,陈伟豪,曾水琴,等.消毒剂对日本鳗鲡肠道N-乙酰-β-D-氨基葡萄糖苷酶活性的影响[J].江苏农业学报,2025,(07):1366-1374.[doi:doi:10.3969/j.issn.1000-4440.2025.07.013]
　LIN Jiancheng,CHEN Weihao,ZENG Shuiqin,et al.Effects of disinfectants on N-acetyl-β-D-glucosaminidase activity of Japanese eel Anguilla japonica[J].,2025,(07):1366-1374.[doi:doi:10.3969/j.issn.1000-4440.2025.07.013]
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消毒剂对日本鳗鲡肠道N-乙酰-β-D-氨基葡萄糖苷酶活性的影响()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2025年07期
页码:
1366-1374
栏目:
畜牧兽医·水产养殖·益虫饲养
出版日期:
2025-07-31

文章信息/Info

Title:
Effects of disinfectants on N-acetyl-β-D-glucosaminidase activity of Japanese eel Anguilla japonica
作者:
林建城; 陈伟豪; 曾水琴; 林娟娟
(莆田学院环境与生物工程学院/福建省新型污染物生态毒理效应与控制重点实验室/生态环境及其信息图谱福建省高等学校重点实验室<莆田学院> ,福建莆田351100)
Author(s):
LIN Jiancheng; CHEN Weihao; ZENG Shuiqin; LIN Juanjuan
(College of Environmental and Biological Engineering, Putian University/Fujian Provincial Key Laboratory of Ecology-toxicological Effects & Control for Emerging Contaminants/Key Laboratory of Ecological Environment and Information Atlas of Fujian Provincial University , Putian 351100, China)
关键词:
消毒剂; 日本鳗鲡; N-乙酰-β-D-氨基葡萄糖苷酶; 抑制作用; 构象变化
Keywords:
disinfectants; Anguilla japonica; N-acetyl-β-D-glucosaminidase; inhibition; conformational change
分类号:
Q556+.2；S965.223
DOI:
doi:10.3969/j.issn.1000-4440.2025.07.013
文献标志码:
A
摘要:
N-乙酰-β-D-氨基葡萄糖苷酶(NAGase)是几丁质酶系主要成员之一,为了探讨消毒剂对养殖鱼体消化道几丁质酶活性的影响,从日本鳗鲡(Anguilla japonica)肠道中分离纯化获得NAGase酶制剂,利用酶促反应动力学和酶抑制作用动力学方法研究10种消毒剂[甲基蓝、高锰酸钾、硫酸铜、氧化钙、甲醛、戊二醛、乙醇、次氯酸钠、二氯异氰尿酸钠和三氯异氰尿酸(TCCA)]对日本鳗鲡NAGase活性的影响；通过对酶荧光发射光谱的测定,研究消毒剂对NAGase蛋白质空间构象的影响。结果表明,在特定质量浓度范围内,甲基蓝和硫酸铜对NAGase活性具有一定的激活作用,高锰酸钾对NAGase有较强的抑制效应，而40.0 mg/L氧化钙可使NAGase活性下降11.79个百分点；甲醛、戊二醛、乙醇、次氯酸钠、二氯异氰尿酸钠和三氯异氰尿酸等对NAGase活性均具有较强的抑制作用。甲醛对NAGase活性的抑制是可逆的,属于非竞争性抑制作用,抑制常数为1.43 mol/L；而TCCA对NAGase的抑制是不可逆的。此外,甲醛可使NAGase蛋白质内源荧光发射强度明显下降,荧光发射峰出现红移。说明甲醛可引起NAGase空间构象变化,也改变了酶中心微环境,导致酶的失活。而小于100 μmol/L浓度的TCCA对NAGase蛋白质内源荧光发射光谱没有明显影响,即TCCA对NAGase空间构象影响很小,可能是通过对酶蛋白质巯基的氧化或与蛋白质氨基的共价结合引起NAGase的不可逆失活。本研究结果证实了10种消毒剂对日本鳗鲡肠道NAGase活性具有较大的调控作用,对水体消毒剂在鱼类养殖中的科学使用具有指导意义。
Abstract:
N-acetyl-β-D-glucosidase (NAGase) is one of the main components of chitinolytic enzymes. In order to investigate the effects of disinfectants on chitinase in the digestive tract of cultured fish, NAGase was extracted from the intestinal tract of Japanese eel, Anguilla japonica. The effects of ten disinfectants including methylene blue, potassium permanganate, copper sulfate, calcium oxide, formaldehyde, glutaraldehyde, ethanol, sodium hypochlorite, sodium dichloroisocyanurate, and trichloroisocyanuric acid (TCCA) on NAGase activities were studied by using the kinetics method of enzyme-catalyzed reaction and kinetics method of enzymatic inhibition. Spatial conformation changes of the NAGase influenced by disinfectants were determined through measuring their fluorescence emission spectrum. The results showed that methlene blue and copper sulfate could activate the enzyme activity, while potassium permanganate had a strong inhibitory effect on the enzyme activity in a certain range of mass concentration. 40.0 mg/L of calcium oxide could lead to losing the enzyme activity by 11.79 percentage points, and formaldehyde, glutaraldehyde, ethanol, sodium hypochlorite, sodium dichloroisocyanurate and TCCA also had strong inhibitory effects on NAGase. The inhibitory effect of formaldehyde on NAGase was reversible and was the non-competitive type. The inhibition constant (Ki) of formaldehyde on the enzyme was determined to be 1.43 mol/L. But the endogenous inhibitory effect of TCCA on NAGase was irreversible. Moreover, the fluorescence emission intensity of the NAGase treated by formaldehyde was decreased obviously and the fluorescence emission peaks were red-shifted. The results indicated that the inactivation effects of formaldehyde on NAGase were caused by the conformational changes and the microenvironment changes in active center of enzyme. But the fluorescence emission spectrum of the NAGase treated by TCCA less than 100 μmol/L concentration had no obvious changes, indicating that TCCA had little effect on the conformation of NAGase. TCCA might oxidate the enzyme’s sulfhydryls or covalently bound to the amino groups of enzyme, leading to the irreversible inactivation on NAGase. The results suggested that ten disinfectants have great regulatory effects on the activities of NAGase in the intestines of Anguilla japonica. The result is instructive for the scientific use of water disinfectants in fish culture.

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备注/Memo

备注/Memo:
收稿日期：2024-11-04基金项目：福建省自然科学基金项目(2023J011004)作者简介：林建城(1966-),男,福建莆田人,硕士,教授,主要从事生物酶学与食品生物技术研究。(E-mail) ptljc660402@sina.com
更新日期/Last Update: 2025-08-19