可口革囊星虫Phasolosma esculenta体腔液的抗菌活性研究

梁倩蓉; 于曙光; 刘顺; 王国良; 郑晓叶; 彭雪美惠; 周素明

doi:10.3969/j.issn.1001-909X.2015.02.010

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海洋学研究 ›› 2015, Vol. 33 ›› Issue (2) : 70-75. DOI: 10.3969/j.issn.1001-909X.2015.02.010

研究论文

可口革囊星虫Phasolosma esculenta体腔液的抗菌活性研究

梁倩蓉, 于曙光, 刘顺, 王国良, 郑晓叶, 彭雪美惠, 周素明^*

作者信息 +

Antibacterial activity of the coelomic fluid from a marine worm, Phasolosma esculenta

LIANG Qian-rong, YU Shu-guang, LIU Shun, WANG Guo-liang, ZHENG Xiao-ye, PENG Xue-mei-hui, ZHOU Su-ming^*

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文章历史 +

摘要

可口革囊星虫Phasolosma esculenta为我国特有的一种可食用海洋经济动物。本文以可口革囊星虫为研究对象,利用琼脂径向扩散、涂板计数等方法研究其体腔液中物质的抗菌活性以及理化特性。研究结果表明,可口革囊星虫体腔液中存在抗菌活性物质,并且该物质对金黄色葡萄球菌Staphylococcus aureus等3种革兰氏阳性细菌具有明显的抗菌效果,但对大肠杆菌Escherichia coli等3种革兰氏阴性细菌以及葡萄牙假丝酵母Candida lusitaniae抗菌效果不明显。结果还发现:(1)不同提取方式出现明显不同的抗菌效果,与体腔液上清液和2%乙酸提取体腔液所得提取物的抗菌效果相比,超声破碎处理提取物的抗菌效果最为明显。(2)理化特性研究发现该抗菌物质抗菌效力持久,抗冻融,并且在5~30 ℃范围内最适抗菌温度约为25 ℃;该物质不耐高温,60 ℃处理10 min后该物质的抗菌作用完全消失;此外,该物质的抗菌作用不依赖金属离子的存在。(3)超滤管过滤截留琼脂扩散结果表明该抗菌物质的分子量在100 kDa以上。

Abstract

Phasolosma esculenta is a peculiar edible marine economic species in China. In this study, the antimicrobial effect and physical and chemical characteristics of coelomic fluid extracted from Phasolosma esculenta was examined with agar diffusion and plate counting method. The result shows that this fluid is found to have an antibacterial activity against three G⁺ bacteria (Staphylococcus aureus, etc), but has no obvious antibacterial activity against three G^- bacteria (Escherichia coli, etc) and the yeast Candida lusitaniae. Results also show that: (1) Different extraction procedure shows different antibacterial activity. Three extraction procedures are compared: (a) the coelomic fluid without any treatment; (b) with ultra sonication and (c) mixed with 2% acetic acid. Pretreated with an ultra-sonication step, the extract has the best antibacterial effect. (2) The antibacterial activity of this component is effective up to 90 minutes and freezing-resistant. With its optimal temperature of 25 ℃, it shows an antibacterial effect within temperature ranging from 5 to 30 ℃. The fluid is inactivated with water bath treatment at 60 ℃ for 10 minutes. In addition, the antibacterial activity is independent from the metal ions. (3) Moreover, by separating fluid components based on molecular weights, it shows that the effective component is larger than 100 kDa.

导出引用

梁倩蓉, 于曙光, 刘顺, 王国良, 郑晓叶, 彭雪美惠, 周素明. 可口革囊星虫Phasolosma esculenta体腔液的抗菌活性研究[J]. 海洋学研究. 2015, 33(2): 70-75 https://doi.org/10.3969/j.issn.1001-909X.2015.02.010

LIANG Qian-rong, YU Shu-guang, LIU Shun, WANG Guo-liang, ZHENG Xiao-ye, PENG Xue-mei-hui, ZHOU Su-ming. Antibacterial activity of the coelomic fluid from a marine worm, Phasolosma esculenta[J]. Journal of Marine Sciences. 2015, 33(2): 70-75 https://doi.org/10.3969/j.issn.1001-909X.2015.02.010

中图分类号： S917.4

参考文献

[1] LI Feng-lu. A checklist of Sipuncula from the China coasts[J]. Journal of Ocean University of Qingdao, 1989,19(3):78-90.
李凤鲁.中国沿海革囊星虫属(星虫动物门)的研究[J].青岛海洋大学学报,1989,19(3):78-90.
[2] JIN Chun-hua, ZHU Jun-quan, XU Shi-jian, et al. Study on the embryonic and larval development of Phascolosoma esculenta[J]. Oceanologia et Limnologia Sinica, 2011,42(1):94-100.
金春华,竺俊全,许式见,等.可口革囊星虫(Phascolosoma esculenta)胚胎及幼虫发育研究[J].海洋与湖沼,2011,42(1):94-100.
[3] DENG Zhong-ri, HUANG Bo, FANG Zai-guang. Morphological and histological characteristics of body wall in Phscolosoma esculenta[J]. Fisheries Science, 2007, 26(6):335-339.
邓中日, 黄勃, 方再光. 可口革囊星虫体壁形态和组织学结构[J]. 水产科学, 2007, 26(6):335-339.
[4] ZHOU Ying-song, DING Li-fa, XU Ji-lin, et al. Analysis of main nutritional components in Phascolosoma esulenta[J]. Acta Nutrimenta Sinica, 2007,29(4):413-414.
周迎松,丁理发,徐继林,等.可口革囊星虫主要营养要素的分析[J].营养学报,2007,29(4):413-414.
[5] ZENG Hai-xiang, ZHU Jun-quan, DING Li-fa. The heavy metal Cadmium and Zinc toxicity test on Phascolosoma esculenta[J]. Reservoir Fisheries, 2006,26(2):96-98.
曾海祥,竺俊全,丁理法.重金属镉和锌对可口革囊星虫的毒性试验[J].水利渔业,2006,26(2):96-98.
[6] DING Li-fa, ZHU Jun-quan, YE Rong-hua, et al. Great artificial breeding of Phascolosoma esculenta[J]. Scientific Fish Farming,2006,2004(5):31-32.
丁理法,竺俊全,叶荣华,等.可口革囊星虫人工养殖获高效[J].科学养鱼,2004(5):31-32.
[7] NIU Rong-li, TANG Jian-hong. Research of anti-fatigue effect of Phascolosoma esculenta[J]. Science and Technology of Food Industry,2012,33(24):389-391.
牛荣丽,唐健红.可口革囊星虫抗疲劳作用的研究[J].食品工业科技,2012,33(24):389-391.
[8] TANG Jing-cai, LIAO Gong-shan, LEI Dan-qing, et al. Study on fibrinolytic enzyme from Phascolosoma esculenta[J]. Journal of SNAKE(Science & NAture are KEy to health),2010,22(4):339-341.
唐景财,廖共山,雷丹青,等.可口革囊星虫体内纤溶酶的初步研究[J].蛇志,2010,22(4):339-341.
[9] LIU Chen, LI Man-xiang, LU Shuai, et al. Progress on antibacterial peptide[J]. Progress in Veterinary Medicine, 2011,32(3):94-99.
刘诚,黎满香,卢帅,等.抗菌肽研究进展[J].动物医学进展,2011,32(3):94-99.
[10] ZHAO Fei. Advances of antimicrobial active substance from Musca domestica( Houseflies)[J]. Journal of Shanxi Agricultural Sciences, 2007,35(9):23-26.
赵飞.家蝇抗菌活性物质研究进展[J].山西农业科学,2007,35(9):23-26.
[11] LIU Yan-qin, WANG Dong-hui, SUN Zhen-jun. In vitro antibacterial effect of earthworm coelomic fluid and crude extracts[J]. Ecology of Domestic Animal,2004,25(4):51-54.
刘艳琴,王东辉,孙振钧.蚯蚓体腔液及粗组分体外抗菌特性[J].家畜生态,2004,25(4):51-54.
[12] DING Yun-chao, ZHANG Shi-cui. Progress on marine animal antimicrobial peptide[J]. Chinese Journal of Marine Drugs,2013,32(6):87-96.
丁云超,张士璀.海洋动物抗菌肽研究进展[J].中国海洋药物,2013,32(6):87-96.
[13] HAUG T, KJUUL A K, STENSVARG K, et al. Antibacterial activity in four marine crustacean decapods[J]. Fish & Shellfish Immunology,2002,12(5):371-385.
[14] CHISHOLMT J R S,SMITH V J. Comparison of antibacterial activity in the hemocytes of different crustacean species[J]. Comparative Biochemistry and Physiology Part A: Physiology,1995,110(1):39-45.
[15] LI C, HAUG T, STYRVOLD O B, et al. Strongylocins, novel antimicrobial peptides from the green sea urchin, Strongylocentrotus droebachiensis[J].Developmental and Comparative Immunology,2008,32(12):1 430-1 440.
[16] CASAS S M, COMESANA P, CAO A, et al. Comparison of antibacterial activity in the hemolymph of marine bivalves from Galicia (NW Spain)[J]. Journal of Invertebrate Pathology, 2011,106(2):343-345.
[17] LANGE S, KAUSCHKE E, MOHRIG W, et al. Biochemical characteristics of Eiseniapore, a pore-forming protein in the coelomic fluid of earthworms[J].European Journal of Biochemistry, 1999, 262(2):547-556.
[18] GUO Dao-sen, WEI Yu-xi, LI Li, et al. Purification of antimicrobial protein from plasma of the mussel Scapharca subcrenata and its antimicrobial activity[J]. Marine Sciences, 2005, 29(3):25-29.
郭道森,魏玉西,李丽,等.毛蚶血浆中抗菌蛋白的纯化及抗菌活性研究[J].海洋科学,2005,29(3):25-29.
[19] BULGAKOV A A, PARK K I, CHOI K S, et al. Purication and characterisation of a lectin isolated from the Manila clam Ruditapes philippinarum in Korea[J]. Fish & Shellfish Immunology,2004,16(4):487-499.
[10] TAKAHASHI K G, KURODA T, MUROGA K. Purification and antibacterial characterization of a novel isoform of the Manila clam lectin (MCL-4) from the plasma of the Manila clam, Ruditapes philippinarum[J]. Comparative Biochemistry and Physiology, Part B, 2008,150(1):45-52.
[21] HUANG Wen-shu, WANG Ke-jian, YANG Ming, et al. Purification and part characterization of a novel antibacterial protein Scygonadin, isolated from the seminal plasma of mud crab, Scylla serrata (Forskl, 1775)[J]. Journal of Experimental Marine Biology and Ecology, 2006,339(1):37-42.
[22] JOHNSON P T, CHAPMAN F A. Comparative studies on the in Vitro response of bacteria to invertebrate body fluids. I.Dendrostomum zostericolum, a sipunculid worm[J]. Journal of Invertebrate Pathology, 1970,16(1):127-138.
[23] KRASSNER S M, FLORY B. Antibacterial factors in the sipunculid worms Golfingia gouldii and Dendrostomum pyroides[J]. Journal of Invertebrate Pathology, 1970,16(3):331-338.
[24] SONG Hong-xia, ZENG Ming-yong, LIU Zun-ying, et al. Biological activity and its mechanism of action of antimicrobial peptides[J]. Science and Technology of Food Industry, 2006,27(9):185-188.
宋宏霞,曾明勇,刘尊英,等.抗菌肽的生物活性及其作用机理[J].食品工业科技,2006,27(9):185-188.