引用本文:	蒋经伟,程雨卉,周遵春,董颖,陈仲,姜北,高杉.光棘球海胆(Strongylocentrotus nudus)酚氧化酶氧化产物的抗菌特性研究*.海洋与湖沼,2015,46(4):969-975.

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光棘球海胆(Strongylocentrotus nudus)酚氧化酶氧化产物的抗菌特性研究*
蒋经伟¹, 程雨卉^1,2, 周遵春¹, 董颖¹, 陈仲¹, 姜北¹, 高杉¹
1.辽宁省海洋水产科学研究院大连 116023;2.大连海洋大学大连 116023

摘要:

本文通过线性连续梯度非变性电泳结合邻苯二酚发色法分离纯化了光棘球海胆(Strongylocentrotus nudus)体腔液上清中的酚氧化酶(PO), 然后通过生长曲线测定法进行了光棘球海胆PO氧化产物的抗菌特性分析。结果显示: (1) 光棘球海胆体腔液上清中存在3种PO (SnPO1, SnPO2, SnPO3), 在聚丙烯酰胺凝胶中与邻苯二酚反应分别呈现褐色、黄色和紫色;(2) 以多巴胺为底物时, SnPO1的氧化产物对灿烂弧菌Vibrio splendidus、哈维氏弧菌Vibrio harveyi和金黄色葡萄球菌Staphylococcus aureus有抗菌作用, SnPO2氧化产物对哈维氏弧菌、金黄色葡萄球菌和拟诺卡式菌Nocardiopsis sp.有抗菌作用, SnPO3氧化产物仅对哈维氏弧菌和拟诺卡式菌有抗菌作用;(3) 以左旋多巴为底物时, 仅SnPO2氧化产物对拟诺卡式菌有抗菌作用;(4) 光棘球海胆PO氧化产物对假交替单胞菌Pseudoalteromonas nigrifaciens、希瓦氏菌Shewanella baltica和溶壁微球菌Micrococcus lysodeikticus的生长无明显影响。上述结果表明, 在光棘球海胆中, PO氧化产物具有窄谱抗菌活性, 多巴胺氧化产物的抗菌活力高于左旋多巴氧化产物, 并且不同PO亚型的氧化产物具有不同的抗菌谱和抗菌特性。

关键词: 光棘球海胆体腔液上清酚氧化酶抗菌特性

DOI：10.11693/hyhz20141200352

分类号:

基金项目:国家863项目,2012AA10A412号;国家自然科学基金项目,31402336号;辽宁省科技计划项目,2014203006号;辽宁省海洋与渔业厅项目,201301号。

附件

ANTIBACTERIAL CHARACTERIZATION OF OXIDATION PRODUCTS GENERATED BY PHENOLOXIDASES FROM THE SEA URCHIN STRONGYLOCENTROTUS NUDUS

JIANG Jing-Wei¹, CHENG Yu-Hui^1,2, ZHOU Zun-Chun¹, DONG Ying¹, CHEN Zhong¹, JIANG Bei¹, GAO Shan¹

1.Liaoning Ocean and Fisheries Science Research Institute, Dalian 116023, China;2.Dalian Ocean University, Dalian 116023, China

Abstract:

We purified three types of phenoloxidase (PO) from the coelomic fluid supernatant of sea urchin Strongylocentrotus nudus (named SnPO1, SnPO2 and SnPO3) using linear-gradient native-PAGE method combined with catechol staining, and studied the antibacterial characteristics of S.nudus PO oxidation products by bacterial growth curve determination.The results show that the three POs presented brown, yellow, and purple, respectively, after reacted with catechol in polyacrylamide gel.With dopamine as substrate, the oxidation products of SnPO1 showed antibacterial activities to Vibrio splendidus, V.harveyi and Staphylococcus aureus, those of SnPO2 to V.harveyi, S.aureus and Nocardiopsis sp., and those of SnPO3 to V.harveyi and Nocardiopsis sp..With L-3, 4-dihydroxyphenylalanine as substrate, only the oxidation products of SnPO2 showed antibacterial activity to Nocardiopsis sp..The oxidation products of S.nudus POs had no obvious effects on the growth of Pseudoalteromonas nigrifaciens, Shewanella baltica and Micrococcus lysodeikticus.Therefore, it could be concluded that S.nudus PO oxidation products had limited antibacterial spectrum, the antibacterial activities in dopamine-derived oxidation products were stronger than those in L-3, 4-dihydroxyphenylalanine-derived oxidation products, and different PO isoforms had different antibacterial spectrum and characteristics.

Key words: Strongylocentrotus nudus coelomic fluid supernatant phenoloxidase antibacterial characteristics