产胞外多糖乳杆菌的筛选及多糖功能的研究  

Screening of Exopolysaccharide-producing Lactobacillus Strains and Study of Exopolysaccharide Properties

作　　者：董阳勤 妥彦峰[1] 牟光庆[1,2] 姜淑娟[1] 钱方[1] 宋莹龙 DONG Yang.-qin TUO Yan-feng MU Guang-qing JIANG Shu-juan QIAN Fang SONG Ying-long(School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China Synergetic Innovation Center of Food Safety and Nutrition, Northeast Agricultural University, Harbin 150030, China)

机构地区：[1]大连工业大学食品学院,辽宁大连116034 [2]东北农业大学食品安全与营养协同创新中心,黑龙江哈尔滨150030

出　　处：《现代食品科技》2017年第2期61-68,共8页Modern Food Science and Technology

基　　金：国家十二五科技支撑计划项目(2013BAD18B09);国家自然科学基金项目(31571813);国家海洋食品工程技术研究中心项目(2012FU125X03);辽宁省自然科学基金项目(2014026018)资助

摘　　要：本论文从大连地区海产品消化腺中筛选产胞外多糖(EPS)的乳杆菌属菌株,并研究其所产EPS的功能特性,选出一株目标菌,对其EPS分离纯化,测定纯化后各多糖组分的相对分子量。获得EPS产量相对较高的菌株经16S r RNA序列测定鉴定为Lactobacillus plantarum subsp.plantarum-4,Lactobacillus plantarum-12,Lactobacillus plantarumsubsp.plantarum-49;对3株植物乳杆菌及其EPS进行清除DPPH自由基的测定,Lactobacillus plantarum-12的EPS浓度为0.5 mg/mL时,清除率为48.82±3.88%,显著高于另外2株菌(p<0.05);测定植物乳杆菌的EPS抑制E.coli ATCC25922在HT-29细胞上的粘附效果,Lactobacillus plantarum-12的EPS浓度为0.2mg/mL时,抑制率为78.23%±2.46%,显著高于另外两株菌(p<0.05);Lactobacillus plantarum-12的EPS可显著抑制E·coli ATCC25922刺激HT-29细胞产生IL-8(p<0.05)。Lactobacillus plantarum-12的EPS经DEAE-Sepharose Fast Flow离子交换柱、Sepharose CL-6B凝胶柱和Sephacryl HR S200凝胶柱层析分离得到两组分,测定两组分的相对分子质量分别为8.5×10~4 u和7.4×10~4 u。Exopolysaccharide（EPS）-producing Lactobacillus strains were screened from the digestive glands of seafood from the Dalian area,and the functional properties of the produced EPSs were studied.One target strain was selected,the corresponding EPS was isolated and purified,and the relative molecular weights of all purified polysaccharide components were measured.Strains with relatively high EPS-producing ability were identified as Lactobacillus plantarum subsp.plantarum-4,Lactobacillus plantarum-12,and Lactobacillus plantarum subsp.plantarum-49.The 2,2-diphenyl-1-picrylhydrazyl（DPPH） free radical scavenging abilities of the three Lactobacillus plantarum strains and their EPSs were measured.When the EPS produced by Lactobacillus plantarum-12 was at a concentration of 0.5 mg/mL,the DPPH free radical scavenging rate was 48.82±3.88%,significantly higher than those of other two strains（P〈0.05）.The effect of EPS produced by Lactobacillus plantarum strains on the inhibition of the adhesion of Escherichia coli ATCC 25922 to HT-29 cells was measured.When the EPS produced by Lactobacillus plantarum-12 was at a concentration of 0.2 mg/ml,the inhibitory rate was 78.23%±2.46%,significantly higher than those of other two strains（P〈0.05）.The EPS produced by Lactobacillus plantarum-12 could significantly inhibit IL-8 production by HT-29 cells stimulated by E.coli ATCC 25922（P〈0.05）.The crude EPS produced by Lactobacillus plantarum-12 was purified by DEAE-Sepharose anion-exchange,Sepharose CL-6B,and Sephacryl HR S200 chromatography columns;two fractions were obtained,and their relative molecular weights were 8.5×10-4 u and 7.4×10-4 u.

关 键 词：胞外多糖 抗氧化性 粘附性 IL-8 纯化 

分 类 号：TS201.3[轻工技术与工程—食品科学]