刺参海带饲料原料褐藻胶降解菌的分离与鉴定  

Isolation and Identification of Algin-Degrading Bacteria in Kelp(Laminaria japonica) Forage Materials for Spiny Sea Cucumber(Apostichopus Japonicas)

作　　者：王熙涛[1,2] 李淑英 王丽丽[3] 李晓宇[3] 徐永平[1,2,3] 

机构地区：[1]动物性食品安全保障技术教育部工程研究中心,辽宁大连116620 [2]大连赛姆生物工程技术有限公司,辽宁大连116620 [3]大连理工大学生命科学与技术学院,辽宁大连116024

出　　处：《微生物学杂志》2016年第6期61-67,共7页Journal of Microbiology

基　　金：国家海洋公益项目(201405003)

摘　　要：从海带及刺参养殖环境中筛选有效降解褐藻胶,且对刺参无致病性的微生物,对海带饲料原料进行降解处理,以降低海带饲料中刺参难以消化的褐藻胶成分,显著提高饲料利用率,增加海带原料价值。以褐藻胶为唯一碳源选择培养基初筛;DNS法测定褐藻胶裂解酶酶活;16S r DNA测序及生理生化试验对菌种进行鉴定;高浓度腹腔攻毒试验考察筛选所得菌株对刺参的潜在致病性;分子排阻色谱及高效凝胶色谱法对微生物酶解褐藻胶的终产物进行分析。系统发育树分析表明,菌株WB1与Bacillus amyloliquifaciens有最高同源性,对刺参无潜在致病性;其褐藻胶裂解酶酶解褐藻胶的终产物主要为二糖和三糖,相对含量分别为74.1%和25.9%,平均分子量为516 Da。解淀粉芽胞杆菌WB1可作为一种安全的有益微生物用于刺参海带饲料原料中褐藻胶成分的降解。An impathogenic microbe that can effectively degrade algin was screen.ed and isolated from kelp and the sea cucumber culturing environment. Using the microbe for degrading and treating kelp forage materials could decrease the algin component that is difficult to digest by spiny sea cucumber （A. japonicas） and increase the utility ratio and the value of kelp forage material. The culture medium with sodium alginate as the sole carbon source was used for preliminary screening; DNS method was used to determine the activity of alginate lyase; 16S rDNA sequencing and physiological and biochemical tests were carried out to characterize the experimental bacterial strain; The strain was investigated by injecting the sea cucumber with a high dose of the bacterial suspension at the coelom to test the toxicity of the microbe for the potential pathogenicity; The end-product of alginate lyase digestion of algin by the strain was analyzed with the method of molecular exclusion chromatography and high gel permeation chromatography. The results showed that the strain WB1 had the highest homology with Bacillus amyloliquefacie~, and had no pathogenicity against spiny sea cucumber, the end products of its algin lysis by lyase mainly were disaccharide and trisaecharide with relative contents at 74. 1% and 25.9% respectively, and average molecular weight at 516 Da. Therefore, the strain B. amyloliquefaciens WB1 can be used as safety, beneficial microbe to lysate the algin component in spiny sea cucumber forage.

关 键 词：刺参 解淀粉芽胞杆菌 海带 褐藻胶 

分 类 号：Q93-331[生物学—微生物学]