海洋微藻的无菌化处理及对其生长特性和生化组成的影响

周文俊; 郑立; 郑明刚; 高伟; 崔志松; 徐鲁燕

海洋微藻的无菌化处理及对其生长特性和生化组成的影响

国家海洋局第一海洋研究所, 海洋生态研究中心, 山东青岛266061

基金项目: 海洋公益性行业专项项目"海洋微藻制备生物柴油的规模化开发利用技术研究"(200805039);国家自然科学基金(41076108;40806053;30870247);青岛市科技发展技术(08-1-3-10-JCH);海洋可再生能源资金项目(GHME2001SW02)。

计量
- 文章访问数: 1701
- HTML全文浏览量: 36
- PDF下载量: 2229
- 被引次数: 0
出版历程
- 收稿日期: 2011-08-10
- 修回日期: 2012-06-15

Axenation of marine microalgae and effects of axenic cultivation on growth characteristics and biochemical compositions of marine microalgae

Research Center for Marine Ecology, the First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China

摘要

摘要: 利用常用抗生素(氨苄青霉素、链霉素、氯霉素和卡那霉素)对4株海洋微藻(1株金藻和3株小球藻)进行了无菌化处理,并对这4株微藻在带菌和无菌培养时的生长特征和生化组成进行了比较。结果表明:中高浓度(≥100 mg/dm³)的单种抗生素或抗生素组合可抑制微藻的生长,个别低浓度(50 mg/dm³)的抗生素或抗生素组合可促进微藻的生长;氨苄青霉素、链霉素和卡那霉素各200 mg/dm³联合使用处理球等鞭金藻,氯霉素100, 50, 50 mg/dm³单独使用分别处理小球藻C95, CV和C146,可获得这4株微藻的无菌藻系;无菌培养的4株海洋微藻的稳定期持续时间由除菌前的4~5 d延长到15~20 d,藻液悬浮性增强,藻细胞不易老化;除菌后微藻的主要生化成分发生明显变化,球等鞭金藻的叶绿素A、叶黄素和总脂含量显著增加,小球藻C95和CV的总蛋白含量显著增加,3株小球藻的脂肪酸组成如EPA, PUFA等的含量发生显著变化。
- 微藻 /
- 无菌化 /
- 抗生素 /
- 生长特征 /
- 生化组成
Abstract: Axenation of four strains of marine microalgae(Isochrysis galbana, Chlorella sp. CV, Chlorella sp. C95 and Chlorella sp. C146) was carried out by using several common antibiotics(including ampicillin, streptomycin, chloramphenicol and kanamycin). The growth characteristics and biochemical compositions of four strains between axenic and nonaxenic cultures were investigated. The results showed: the growth of microalgae would be inhibited if the concentrations of single antibiotic or combined antibiotics were equal or higher than 100 mg/L, while the growth would be stimulated if the concentrations of some single or combined antibiotics were at 50mg/L; the axenic culture of I. galbana was obtained by combinedly using 200 mg/L of ampicillin, streptomycin and kanamycin, and the axenic cultures of C. CV, C. 95 and C. C146 were obtained by singly using 100, 50 and 50 mg/L of chloramphenicol respectively; the duration of stationary phase of the four axenic cultures prolonged for about 15 days, and the axenic algae could maintain a good suspension property rather than began aging prematurely; the contents of chlorophyll A, lutein and total lipids in axenic I. galbana and total protein in axenic C. CV and C. 95 increased significantly, and the composition of fatty acids such as EPA and PUFA in axenic Chlorella species changed markedly.
- microalgae /
- axenation /
- antibiotic /
- growth characteristics /
- biochemical compositions

HTML全文

参考文献(38)

SHI X M, ZHANG X W, CHEN F. Heterotrophic production of biomass and lutein by Chlorella protothecoides on various nitrogen sources[J]. Enzyme and Microbial Technology, 2000, 27(3/5): 312-318.

WEN Z Y, CHEN F. Heterotrophic production of eicosapentaenoid acid by the diatom Nitzschia laevis: effects of silicate and glucose[J]. Journal of Industrial Microbiology & Biotecnology, 2000, 25(4): 218-224.

MIAO X L, WU Q Y. Biodiesel production from heterotrophic microalgal oil[J]. Bioresource Technology, 2006, 97(6): 841-846.

PAHL S L, LEWIS D M, CHEN F, et al. Heterotrophic growth and nutritional aspects of the diatom Cyclotella cryptica(Bacillariophyceae): Effect of some environmental factors[J]. Journal of Bioscience and Bioengineering, 2010, 109(3): 235-239.

林伟,陈騳,刘秀云.海洋微藻除菌及除菌与自然带菌微藻生长特点比较[J].海洋与湖沼,2000,31(6):647-652.

BOWYER J W, SKERMAN V B D. Production of axenic cultures of soil-borne and endophytic blue-green algae[J]. Journal of General Microbiology, 1968, 54(2): 299-306.

汪本凡,赵良侠,叶霁,等.微藻无菌化技术的研究进展[J].微生物学通报,2007,34(2):363-366.

AL-AHMAD A, DASCHNER F D, KVMMERER K. Biodegradability of cefotiam, ciprofloxacin, meropenem, penicillin G, and sulfamethoxazole and inhibition of waste water bacteria[J]. Archives of Environmental Contamination and Toxicology, 1999, 37(2): 158-163.

SPENCER C P. On the use of antibiotics for isolating bacteria-free cultures of marine phytoplankton organisms[J]. Journal of the Marine Biological Association of the United Kingdom, 1952, 31(1): 97-106.

JONES A K, RHODES M E, EVANS S C. The use of antibiotics to obtain axenic cultures of algae[J]. European Journal of Phycology, 1973, 8(2): 185-196.

DIVAN C L, SCHNOES H K. Production of axenic Gonyaulax cultures by treatment with antibiotics[J]. Applied and Environmental Microbiology, 1982, 44(1): 250-254.

COTTRELL M T, SUTTLE C A. Production of axenic cultures of Micromonas pusilla(Prasinophyceae) using antibiotics[J]. Journal of Phycology, 1993, 29(3): 385-387.

林伟.几种海洋微藻的无菌化培养[J].海洋科学,2000,24(10):4-6.

王江涛,尹晓楠,宋茜.几种抗生素的抑菌效果及其对中肋骨条藻生长的影响[J].中国海洋大学学报,2008,38(3):468-472.

周文礼,乔秀亭,肖慧,等.三种抗生素对几种海洋微藻叶绿素a含量影响的初步研究[J].海洋环境科学,2009,28(3):268-271.

郝雯瑾,王悠,唐学玺.除菌处理对强壮前沟藻和青岛大扁藻生长的影响[J].应用与环境生物学报,2009,15(3):326-331.

BOROWITZKA M A, BOROWITZKA L J. Micro-algal Biotechnology[M]. Cambridge: Cambridge University Press, 1988: 457-465.

程红艳,陈军辉,张道来,等.超声波辅助提取RP-HPLC法测定浒苔中的叶绿素a、b[J].海洋科学,2010,34(2):23-27.

程红艳,陈军辉,赵恒强,等.反相高效液相色谱-电喷雾质谱法测定浒苔中叶黄素[J].食品科学,2010,31(18):206-211.

BLIGH E G, DYER W J. A rapid method of total lipid extraction and purification[J]. Canadian Journal of Biochemistry and Physiology, 1959, 37(8): 911-917.

CARVALHO A P, MALCATA F X. Preparation of fatty acid methyl esters for gas-chromatographic analysis of marine lipids: insight studies[J]. Journal of Agricultural and Food Chemistry, 2005, 53(13): 5049-5059.

RASOUL-AMINI S, GHASEMI Y, MOROWVAT M H, et al. PCR amplification of 18S rRNA, single cell protein production and fatty acid evaluation of some naturally isolated microalgae[J]. Food Chemistry, 2009, 116(1): 129-136.

赵培,王雪青,朱潮峰,等.3种常用抗生素应用于海洋微藻无菌化培养的研究[J].天津师范大学学报(自然科学版),2007,27(2):27-30.

FAUST M, ALTENBURGER R, BACKHAUS T, et al. Predictive assessment of the aquatic toxicity of multiple chemical mixtures[J]. Journal of Environmental Quality, 2000, 29(4): 1063-1068.

BACKHAUS T, SCHOLZE M, GRIMME L H. The single substance and mixture toxicity of quinolones to the bioluminescent bacterium Vibrio fischeri[J]. Aquatic Toxicology, 2000, 49(1/2): 49-61.

URIARTE I, FARÍAS A, CASTILLA J C. Effect of antibiotic treatment during larval development of the Chilean scallop Argopecten purpuratus[J]. Aquacultural Engineering, 2001, 25(3): 139-147.

周文礼,王悠,肖慧,等.不同海洋饵料微藻对抗生素的敏感性差异分析[J].武汉大学学报(理学版),2007,53(2):249-254.

SAHUL-HAMEED A S, BALASUBRAMANIAN G. Antibiotic resistance in bacteria isolated from Artemia nauplii and efficacy of formaldehyde to control bacterial load[J]. Aquaculture, 2000, 183(3/4): 195-205.

TORKILDSEN L, MAGNESEN T. Hatchery production of scallop larvae(Pecten maximus)-survival in different rearing systems[J]. Aquaculture International, 2004, 12(4/5): 489-507.

杨世平,刘慧玲,李活,等.两种微藻在无菌和带菌状态下生长特点比较研究[J].渔业现代化,2009,36(5):45-49.

MAYALI X, DOUCETTE G J. Microbial community interactions and population dynamics of an algicidal bacterium active against Karenia brevis(Dinophyceae)[J]. Harmful Algae, 2002, 1(3): 277-293.

龚良玉,王修林,李雁宾,等.铜绿假单胞菌产吩嗪类色素的分离纯化及其对赤潮生物生长的影响[J].复旦学报(自然科学版),2004,43(4):494-499.

AMARO A M, FUENTES M S, OGALDE S R, et a1. Identification and characterization of potentially algal-lytic marine bacteria strongly associated with the toxic dinoflagellate Alexandrium catenella[J]. Journal of Eukaryotic Microbiology, 2005, 52(3): 191-200.

VOLK R B. Screening of microalgal culture media for the presence of algicidal compounds and isolation and identification of two bioactive metabolites, excreted by the cyanobacteria Nostoc insulare and Nodularia harveyana[J]. Journal of Applied Phycology, 2005, 17(4): 339-347.

郑立,韩笑天,严小军,等.海洋生物共栖细菌抑藻活性的初步研究[J].海洋科学进展,2006,24(4):511-519.

SANDERS R W, CARON D A, DAVIDSON J M, et al. Nutrient acquisition and population growth of a mixotrophic alga in axenic and bacterized cultures[J]. Microbial Ecology, 2001, 42(4): 513-523.

徐金森,郑天凌,郭清华,等.两种海洋细菌对赤潮藻的细胞生物量的影响研究[J].海洋科学,2002,26(12): 57-60.

王金娜,严小军,周成旭,等.产油微藻的筛选及中性脂动态积累过程的检测[J].生物物理学报,2010,26(6):472-480.

施引文献

资源附件(0)

访问统计