产胞外多糖极地微生物的筛选及其胞外多糖对大菱鲆免疫活力的初步研究  

Screening of Exopolysaccharide-producing Antarctic Bacteria and the Preliminary Study of Effects of EPSs on Nonspecific Immunity of Turbot (Scophthalmus maximus)

作　　者：赵惠娅[1,2] 刘同军[1] 郑风荣[2] 林学政[2] 

机构地区：[1]山东轻工业学院食品与生物工程学院,山东济南250353 [2]国家海洋局第一海洋研究所国家海洋局海洋生物活力物质重点实验室,山东青岛266061

出　　处：《海洋科学进展》2013年第2期237-246,共10页Advances in Marine Science

基　　金：海洋公益性行业研究专项--极地微生物免疫活性多糖的研制及其在海水养殖中的应用技术研究(20105020)

摘　　要：采用苯酚硫酸法和醇沉淀法,从600余株南极微生物中筛选得到10株胞外多糖产量较高的菌株。分子鉴定与系统发育分析表明,9株菌株属于假交替单胞菌属(Pseudoalteromonas),1株属于嗜冷杆菌属(Psychrobacter)。对10株菌株进行发酵培养,发酵液经醇沉、除蛋白、冷冻干燥后得到粗胞外多糖,其中菌株3-3-1-2的胞外多糖产量达0.6g/L。对粗多糖产量较高的4株菌所产的胞外多糖进行免疫活性试验,粗胞外多糖20#3在注射3d时,实验组AKP活性比对照组增加了39%,CAT活性比对照组增加了24%,二者均差异显著(P<0.05),注射5d时,ACP活性比对照组增加了20%,差异显著(P<0.05),总超氧化物歧化酶(T-SOD)活性比对照组增加了14%,差异极显著(P<0.01);粗胞外多糖3-3-1-2在注射3d时,实验组AKP活性比对照组增加了12%,差异不显著(P>0.05),注射5d时,酸性磷酸酶(ACP)活性比对照组增加了24%,过氧化氢酶(CAT)活性比阳性对照组增加了5%,二者均差异显著(P<0.05)。研究结果表明,菌株20#3与3-3-1-2产生的胞外多糖对大菱鲆(Scophthalmus maxoi-mus)具有显著的免疫促进作用。由此可知,筛选出的极地细菌胞外多糖20#3和3-3-1-2可显著提高大菱鲆非特异性免疫力,可作为水产饲料添加剂应用于大菱鲆的生产养殖。Ten Antarctic bacteria strains with higher exopolysaccharide-producing ability were screened from more than 600 strains by phenol-sulfuric acid and alcohol precipitation. Phylogenetic analysis showed that 9 strains belonged to genus of Pseudoalteromonas and 1 strain belonged to genus of Psyehrobacter. The crude exopolysaecharide （EPS） was obtained after fermentation broth treated by alcohol precipitation, proteins removal （Sevag method） and lyophilized. Strain 3-3-1-2 had highest EPS-producing ability, and its EPS yield was up to 0. 6 g/L. The EPS produced by 4 strains with higher yield was chosen to test their immunologic activity on turbot. Results showed that on the 3 d after injected with EPS 20 #3, the activi- ties of both AKP and CAT in the serum of turbot increased 39% and 24% than those of control groups re- spectively, the differences were significant （P〈0.05）. On the 5 d after injection, the activity of ACP in creased 20% than that of control group, the difference was significant（P%0.05）. The activity of T-SOD increased 24 % than that of control group, the increase was extremely significant than that of control group （P%0.01）. On the 3 d after injected with EPS 3-3-1-2, the activity of AKP in the serum of turbot in- creased 12% than that of control group, the increase was not significant（P〉0.05）. On the 5 d after injec tion, the activity of both ACP and CAT increased 24% and 5% than those of positive control groups re spectively, the increases were significant higher（P〈0.05）. All these results indicated that 2 of 4 kinds of EPS had significant effects on immunologic activity of turbot. It indicates that EPS produced by strain 20 3 and 3-3-1-2 had significant effects on the nonspecific immunity index of turbot, and they could be used as additives and immunity-stimulants in the aquaculture industry of turbot.

关 键 词：极地微生物 胞外多糖 大菱鲆 免疫活力 

分 类 号：Q939.91[生物学—微生物学]