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玉米赤霉烯酮降解菌的分离、鉴定及其适宜降解条件的研究
引用本文:金博文,徐长春,李根,李晓宇,王丽丽,李纪彬,徐永平.玉米赤霉烯酮降解菌的分离、鉴定及其适宜降解条件的研究[J].中国畜牧兽医,2020,47(9):2808-2815.
作者姓名:金博文  徐长春  李根  李晓宇  王丽丽  李纪彬  徐永平
作者单位:1. 大连理工大学生物工程学院, 大连 116024;2. 大连金普新区农业农村发展服务中心, 大连 116100;3. 大连赛姆生物工程技术有限公司, 大连 116620;4. 大连赛姆生物工程技术有限公司博士后工作站, 大连 116620
基金项目:国家自然科学基金国际(地区)合作与交流项目(41861124004)
摘    要:试验旨在利用微生物降解法对饲料中存在的玉米赤霉烯酮(zearalenone,ZEN)进行快速、有效的消减。从不同地区采集了牛粪、羊粪及土壤样品共70份,并从中分离出164株菌株。利用环戊酮作为唯一碳源对这164株菌株进行初筛,再以ZEN作为唯一碳源对初筛菌株进行复筛并获得了8株ZEN降解菌株。采用高效液相色谱法(HPLC)对复筛得到的8株ZEN降解菌株进行ZEN降解率的测定,得到1株降解效果良好的菌株即NF-PJ-5号菌株。该菌株经16S rDNA序列比对、构建系统发育树、菌落形态观察及生理生化试验鉴定后确定为多食鞘氨醇杆菌(Sphingobacterium multivorum)。分别研究该菌株在不同温度、不同pH及不同菌液浓度条件下对ZEN的降解能力,评估菌株性质;同时,初步探究该菌株对ZEN的清除机理。结果显示,在28~32 ℃,pH 6.5~7.5,菌液浓度在2×109 CFU/mL以上时,该菌株对ZEN保持较高的降解率。在30 ℃、pH 7.0、160 r/min培养48 h后,该菌株对初始浓度为10 μg/mL的ZEN降解率达到83%。综上,本试验筛选到1株ZEN降解能力较强的菌株,经鉴定为多食鞘氨醇杆菌。初步推测该菌株可通过产生胞外酶降解ZEN,同时该菌细胞壁对ZEN具有一定吸附能力。

关 键 词:玉米赤霉烯酮  微生物降解  多食鞘氨醇杆菌  鉴定  
收稿时间:2020-02-27

Isolation,Identification and Its Suitable Degradation Conditions of a Zearalenone-degrading Bacteria
JIN Bowen,XU Changchun,LI Gen,LI Xiaoyu,WANG Lili,LI Jibin,XU Yongping.Isolation,Identification and Its Suitable Degradation Conditions of a Zearalenone-degrading Bacteria[J].China Animal Husbandry & Veterinary Medicine,2020,47(9):2808-2815.
Authors:JIN Bowen  XU Changchun  LI Gen  LI Xiaoyu  WANG Lili  LI Jibin  XU Yongping
Institution:1. School of Bioengineering, Dalian University of Technology, Dalian 116024, China;2. Center for Agricultural and Rural Development and Service, Jinpu New District of Dalian, Dalian 116100, China;3. Dalian SEM Bio-Engineering Technology Co., Ltd., Dalian 116620, China;4. Post-Doctoral Workstation, Dalian SEM Bio-Engineering Technology Co., Ltd., Dalian 116620, China
Abstract:The aim of this study was to reduce zearalenone (ZEN) in feed by bio-degradation rapidly and effectively.A total of 70 samples of cow manure,sheep manure and soil were collected from different areas and then 164 strains were isolated from these samples.Cyclopentanone was used as the only carbon source to preliminarily screen the 164 strains isolated from the collected samples,and the second-screening was performed using zearalenone as the only carbon source.As a result,8 ZEN-degrading strains were obtained.The ZEN-degrading efficiency of 8 strains was determined by high performance liquid chromatography (HPLC),and a strain with high ZEN-degrading,namely NF-PJ-5 strain.The strain was identified as Sphingobacterium multivorum by 16S rDNA sequence comparison,phylogenetic tree construction,colony morphology observation and physiological and biochemical identification.The ability of NF-PJ-5 strain to degrade zearalenone at different temperature,different pH and different concentration of bacterial solution were studied.In addition,the mechanism of ZEN degradation by this strain was studied.The results showed that at 28-32 ℃,pH 6.5-7.5,and the concentration of bacterial solution above 2×109 CFU/mL,the strain could maintain a high degradation rate.Besides,the strain could degrade 83% ZEN with an initial concentration of 10 μg/mL,after 48 h culture at 30 ℃,pH 7.0,and a shaking speed of 160 r/min.From the above,a highly efficient ZEN-degrading strain was screened in this study,and it was identified as Sphingobacterium multivorum.It was preliminarily speculated that the strain could degrade ZEN by producing extracellular enzyme,and the cell wall of the strain had a certain adsorption capacity for ZEN.
Keywords:zearalenone  bio-degradation  Sphingobacterium multivorum  identification  
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