降解脱氧雪腐镰刀菌烯醇菌株的筛选与鉴定  

Screening and Identification of Strains Degrading Deoxynivalenol

作　　者：王俊峰[1] 耿凯伟 刘宇翾 时子瑶 宫玉杰 田亚东[2] 康相涛[2] 王彦彬[1] 孙向丽[2] WANG Junfeng;GENG Kaiwei;LIU Yuxuan;SHI Ziyao;GONG Yujie;TIAN Yadong;KANG Xiangtao;WANG Yanbin;SUN Xiangli(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China;College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物医学院,郑州450046 [2]河南农业大学动物科技学院,郑州450046

出　　处：《动物营养学报》2023年第8期5430-5440,共11页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：河南省高校科技创新团队支持计划(21IRTSTHN022);中原科技创新领军人才项目(214200510003)。

摘　　要：本试验旨在筛选能够降解脱氧雪腐镰刀菌烯醇(DON)的微生物,应用于DON的脱毒。试验以DON为唯一碳源,从土壤、鸡和猪粪便等样品中筛选可以降解DON的菌株。选取其中降解率最高的菌株,通过细菌形态学、生理生化特征、16S rRNA以及特异性基因gyrB序列测定进行菌株鉴定,并探究其降解DON活性成分,以及DON的降解产物对猪小肠上皮细胞(IPEC-J2细胞)活力和凋亡的影响。结果表明:1)从小麦田土壤样品中分离出了1株能够降解DON(1μg/mL)的菌株,在37℃培养96 h后对DON的降解率为60.77%,经鉴定并命名为解淀粉芽孢杆菌H7(Bacillus amyloliquefaciens H7)。2)解淀粉芽孢杆菌H7菌株主要通过分泌胞外酶降解DON。3)在CCK-8试验中,处理IPEC-J2细胞24和48 h后,0.5和1.0μg/mL DON降解产物组细胞存活率极显著高于DON组(P<0.01);在细胞凋亡试验中,处理IPEC-J2细胞24和48 h后,0.5μg/mL DON降解产物组细胞早期凋亡率和总凋亡率都显著低于DON组(P<0.05)。综上所述,本试验筛选到1株可降解DON的解淀粉芽孢杆菌H7,其活性物质胞外酶主要分布在细菌培养液上清中;DON经降解后对猪小肠上皮细胞的毒性降低。The aim of this study was to screen the microorganisms capable of degrading deoxyniniolenol(DON)for use in the detoxification of DON.The strains that degrade DON were screened from the soil,chicken and pig manure samples using DON as the sole carbon source.The strains with the highest degradation rates were selected and identified by morphology,physiological and biochemical characteristics,16S rRNA and specific gyrB sequences,and investigated the effects of the active components of DON degradation and the deg⁃radation products of DON on the viability and apoptosis of porcine small intestinal epithelial cells(IPEC⁃J2).The results showed as follows:1)a strain capable of degrading DON(1.0μg/mL)was isolated from wheat field soil samples,and the degradation rate was 60.77%after 96 h incubation at 37℃.It was identified and named Bacillus amyloliquefaciens H7.2)Bacillus amyloliquefaciens strain H7 degraded DON mainly by secre⁃ting extracellular enzymes.3)In the CCK⁃8 assay,the survival rates of the 0.5 and 1.0μg/mL DON degrada⁃tion product groups were extremely significantly higher than those of the DON group after 24 and 48 h of treat⁃ment of IPEC⁃J2 cells(P<0.01);in the apoptosis assay,after treating IPEC⁃J2 cells for 24 and 48 h,the ear⁃ly apoptosis rate and the total apoptosis rate were significantly lower in the 0.5μg/mL DON degradation prod⁃uct group than in the DON group(P<0.05).In summary,this assay identified a strain of Bacillus amylolique⁃faciens H7 that efficiently degraded DON,with its active substance extracellular enzyme mainly distributed in the culture supernatant of the bacteria;the toxicity of DON to porcine small intestinal epithelial cells is reduced after degradation.

关 键 词：解淀粉芽孢杆菌 脱氧雪腐镰刀菌烯醇 酶降解 细胞毒性 

分 类 号：S816[农业科学—饲料科学]