研究简报 Short Communications

山羊瘤胃中单宁降解菌的分离、筛选和鉴定

展开
  • 1. 广西大学动物科学技术学院, 南宁 530005;
    2. 南宁学院, 南宁 530200
李兴芳(1962—),女,壮族,广西南宁人,高级实验师,主要从事生物技术在饲料中应用的研究。E-mail: 105321254@qq.com

收稿日期: 2015-01-05

  网络出版日期: 2015-06-17

基金资助

南宁市科学研究与技术开发计划(201102048B)

Isolation, Screening and Identification of Tannin Degradable Bacteria in Rumen of Goats

Expand
  • 1. Animal Science and Technology of Guangxi University, Nanning 530005, China;
    2. Nanning University, Nanning 530200, China

Received date: 2015-01-05

  Online published: 2015-06-17

摘要

本试验旨在从山羊瘤胃中分离、筛选和鉴定单宁降解菌。从隆林山羊瘤胃采集内容物,采用单宁浓度耐受培养基进行初筛,再采用单宁酶活力鉴定培养基分离、筛选单宁降解菌,筛选出的细菌通过16S rDNA基因序列测定初步鉴定,将菌株在单宁浓度为0.5%、1%、1.5%的单宁酶诱导培养基中培养,测定酶活力。结果表明:共分离、筛选到4株降解单宁的细菌,根据16S rDNA基因序列分析结果,分别为产碱普罗威登斯菌(Providencia alcalifaciens)、克斯特菌(Kerstersia)、普罗威登斯菌(Providencia vermicola)和铜绿假单胞菌(Pseudomonas aeruginosa)。经单宁酶诱导培养基培养后,Kerstersia的胞内单宁酶活力高于其他3株菌,而其他3株菌的酶活力较为接近;KerstersiaProvidencia vermicolaPseudomonas aeruginosa经1%单宁浓度诱导的单宁酶活力均高于经0.5%和1.5%单宁浓度诱导(P<0.05)。本试验成功从山羊瘤胃中筛选、分离、鉴定出4株单宁降解菌,且它们均表现出较高的单宁酶活力。

本文引用格式

李兴芳, 徐雯, 张胜男, 徐高跷, 汪珊如, 王士长, 梁世忠 . 山羊瘤胃中单宁降解菌的分离、筛选和鉴定[J]. 动物营养学报, 2015 , 27(6) : 1921 -1927 . DOI: 10.3969/j.issn.1006-267x.2015.06.031

Abstract

This study was conducted to isolate, screen and identify several strains of tannin degradable bacteria in rumen of goats. Ruminal contents was collected from goats, and were preliminary screened by tolerance culture medium with different tannic concentrations, then strains of tannin degradable bacteria were isolated, screened by tannase identification culture mediums. The screened strains were firstly identified by the methods of 16S rDNA gene sequencing, and then were cultured on tannase inductive culture mediums with different tannin concentrations (0.5%, 1% and 1.5%, respectively) for the analysis of enzyme activity. The results showed as follows: four strains of tannin degradable bacteria were isolated and screened. According to the results of 16S rDNA gene sequencing, the strains were Providencia alcalifaciens, Kerstersia, Providencia Vermicola and Pseudomonas aeruginosa, respectively. After cultured on tannase inductive culture medium, the intra-cellular tannase activity of Kerstersia was higher than that of the other three strains, which was pretty close to each other; after inducted by 1.5% tannin, the activity was significantly higher than that inducted by 0.5% and 1% tannin (P<0.05). In conclusion, four strains with high enzyme activity of tannin degradable bacteria in rumen of goats was successfully isolated, screened and identified.

参考文献

[1] FRAZIER R A,DEAVILLE E R,GREEN R J,et al.Interactions of tea tannins and condensed tannins with proteins[J].Journal of Pharmaceutical and Biomedical Analysis,2010,51(2):490-495.  

[2] GETACHEW G,PITTROFF W,PUTNAM D H,et al.The influence of addition of gallic acid,tannic acid,or quebracho tannins to alfalfa hay on in vitro rumen fermentation and microbial protein synthesis[J].Animal Feed Science and Technology,2008,140(3/4):444-461.

[3] LI M S,YAO K,HE Q,et al.Biodegradation of gallotannins and ellagitannins[J].Journal of Basic Microbiology,2006,46(1):68-84.  

[4] FIELD J A,LETTINGA G.Toxicity of tannic compounds to microorganisms[M]//HEMINGWAY R W,LAKES P E.Plant Polyphenols.New York:Plenum Press,1992:673-692.

[5] GOLDSTEIN J L,SWAIN T.The inhibition of enzymes by tannins[J].Phytochemistry,1965,4(1):185-192.  

[6] CLOUTIER M M,GUERNSEY L.Tannin inhibits adenylate cyclase in airway epithelial cells[J].The American Journal of Physiology,1995,268(5):815-855.

[7] OH H,HOFF J E.Effect of condensed grape tannins on the in vitro activity of digestive proteases and activation of their zymogens[J].Food Science,1986,51(3):577-580.  

[8] SKENE I K,BROOKER J D.Characterization of tannin acylhydrolase activity in the ruminal bacterium,Selenomonas ruminantium[J].Anaerobe,1995,1(6):321-327.  

[9] ODENYO A A,MCSWEENEY C A,PALMER B,et al.In vitro screening of rumen fluid samples from indigenous African ruminants provides evidence for rumen fluid with superior capacities to digest tannin-rich fodders[J].Australian Journal of Agricultural Research,1999,50(7):1147-1157.  

[10] GOEL G,PUNIYA A K,SINGH K.Phenotypic characterization of tannin-protein complex degrading bacteria from faeces of goat[J].Small Ruminant Research,2007,69(1/2/3):217-220.

[11] SINGH B,BHAT T K,SHARMA O P,et al.Isolation of tannase-producing Enterobacter ludwigii GRT-1 from the rumen of migratory goats[J].Small Ruminant Research,2011,102(2/3):172-176.

[12] 张武先,王金华,熊智,等.思茅松毛虫2龄幼虫肠道好氧细菌的筛选及毒力测定[J].安徽农业科学,2011,39(28):17288-17290.

[13] BRADOO S,GUPTA R,SAXENA R K.Screening of extracellular tannase-producing fung:development of a rapid and simple plate assay[J].Journal of General and Applied Microbiology,1996,42(4):325-329.  

[14] 郭鲁宏.单宁酶的制备及酶法制取没食子酸及其丙酯研究[D].博士学位论文.沈阳:中国科学院沈阳应用生态研究所,1999.

[15] 保玉心.固体发酵黑曲霉产单宁酶的研究[D].硕士学位论文.贵阳:贵州大学,2008.

[16] 许彬.米曲霉Aspergillus oryzae中单宁酶基因的克隆及其在大肠杆菌和毕赤酵母中的表达[D].硕士学位论文.南宁:广西大学,2003.

[17] 黄小凤,韦宇拓,韦传东,等.单宁酶基因在黑曲霉ST31中的克隆与表达[J].中国生物工程杂志,2005,25(9):74-77.

[18] NOGUCHI N,OHASHI T,SHIRATORI T,et al.Association of tannase-producing Staphylococcus lugdunensis with colon cancer and characterization of a novel tannase gene[J].Journal of Gastroenterology,2007,42(5):346-351.  

[19] RODRÍGUEZA H,DE LAS RIVASA B,GÓMEZ-CORDOVÉSB C,et al.Characterization of tannase activity in cell-free extracts of Lactobacillus plantarum CECT 748T[J].International Journal of Food Microbiology,2008,121(1):92-98.  
文章导航

/