白芥子诱导的鸡内源性芥子碱降解菌的筛选、鉴定及酶活测定  

Screening,identification and enzyme activity determination of chicken endogenous sinapine-degrading bacteria induced by semen brassicae

作　　者：邓卉[1] 余丹[1] 范景胜[1] 屈东[1] 邹成义[1] 李斌[1] 殷勤[1] 郑钰嘉 倪青松[1] 汪林书[1] DENG Hui;YU Dan;FAN Jingsheng;QU Dong;ZOU Chengyi;LI Bin;YIN Qin;ZHENG Yujia;NI Qingsong;WANG Linshu(Sichuan Animal Science Academy,Animal Breeding and Genetics Key Laboratory of Sichuan Province,Chengdu,Sichuan Province 610066,China)

机构地区：[1]四川省畜牧科学研究院,动物遗传育种四川省重点实验室,四川成都610066

出　　处：《中国饲料》2023年第3期44-49,共6页China Feed

基　　金：四川省公益性科研院所基本科研项目(SASA202107);四川省公益性科研院所基本科研项目(SASA202204)。

摘　　要：本试验旨在通过分离鉴定白芥子诱导的芥子碱降解菌,并对其核心功能酶漆酶进行酶活测定,建立芥子碱的生物降解方法,为探寻白芥子等十字花科类非粮饲料资源的高效利用提供技术支撑。本试验以藏香鸡为模型,在基础日粮中添加20%白芥子饲喂28 d进行芥子碱降解菌体内诱导。分别采集鸡空肠、回肠、盲肠内容物在不同培养基中分离筛选芥子碱降解菌,对筛选出的优势菌进行形态学和分子生物学鉴定,测定该菌株的芥子碱降解效果和漆酶活性。结果表明:(1)在鸡盲肠中筛选出9株目标菌,筛出率明显高于空肠和回肠,并进一步选出1株芥子碱降解优势菌SDB2(Sinapine-degrading bacteria 2)。(2)经鉴定,SDB2菌株属于变形菌门;γ-变形菌纲;肠杆菌目;肠杆菌科;大肠埃希菌。(3)芥子碱浓度与SDB2菌株培养时间存在极显著的线性回归关系(P <0.01),芥子碱浓度随培养时间增加而降低,培养72 h的芥子碱降解率可达50%。(4)漆酶活力与SDB2菌株培养时间存在极显著的线性回归关系(P <0.01),漆酶活力随培养时间增加而降低。(5)漆酶活力与芥子碱浓度存在极显著的线性回归关系(P <0.01),漆酶活力与芥子碱浓度呈正相关。综上,本试验条件下,筛选出的SDB2能利用自身分泌的漆酶有效降解芥子碱,为高效利用十字花科类非粮饲料资源奠定了基础。To explore the biodegradation method of sinapine and utilize the non-food feed resources of Brassicaceae such as semen brassicae efficiently,this study isolated culture and identified a sinapine-degrading bacteria induced by semen brassicae,determined the laccase activity of the bacteria. Tibetan chickens were fed with the broiler commodity rations which was mixed with 20% semen brassicae in 28 days.Contents of jejunum,ileum and cecum in the broilers was cultivated in different media respectively to separated sinapine-degrading bacteria.Identified the dominant bacteria by morphological and molecular biological methods,measured the dominant bacteria in degradation rate of sinapine and the laccase activity of the strain.The results showed as follows:(1)9 strains of target bacterium were isolated from the cecum of Tibetan chickens.It seemed that it was easier to isolate sinapine-degrading bacterium from the cecum than that from the jejunum and ileum in Tibetan chickens,and a dominant strain of sinapine-degrading bacteria was identified as SDB2(sinapine-degrading bacteria 2).(2)Strain SDB2 was identified as Proteobacteria;γ-proteobacteria;Enterobacterales;Enterobacteriaceae;Escherichia.(3)There was a very significant linear regression between the concentration of sinapine and the culture time of SDB2 strain(P < 0.01).The concentration of sinapine decreased with the increase of culture time,and the degradation rate of sinapine was up to 50%.(4) There was a very significant linear regression between laccase activity and culture time of SDB2 strain(P < 0.01),and the laccase activity decreased with the increase of culture time.(5)There was a very significant linear regression relationship between laccase activity and sinapine concentration(P < 0.01),and laccase activity was positively correlated with sinapine concentration.Under the conditions of this experiment,SDB2 could degrade sinapine by its laccase effectively and lay a foundation for the efficient utilization of non-food feed resources.

关 键 词：芥子碱降解菌 芥子碱 漆酶 白芥子 大肠杆菌 

分 类 号：S816.3[农业科学—饲料科学]