为获得良好的产生物胺氧化酶的肉品发酵剂,本试验以新疆不同地区的44种熏马肠样品为研究对象,对产生物胺氧化酶菌进行筛选和鉴定,利用单层培养、双层显色的方法剔除产生物胺菌株,对分离后得到的菌株进行氧化酶试验、抑菌试验、耐盐和耐亚硝酸盐试验。采用DGGE剔除重复菌株,通过HPLC对所筛选菌株氧化生物胺能力进行验证,最后对筛选出的菌株提取DNA,并对其16S rDNA的V6-V8区段进行PCR扩增后测序。结果表明:本次试验共筛选出6株菌有氧化酶特性,耐盐能力在6%以上,耐亚硝酸盐能力在0.15 g/kg以上,对致病性大肠杆菌,单增李斯特菌,金黄色葡萄球菌三种致病菌有良好的抑制效果,经HPLC验证后氧化生物胺能力较强,分别为鼠李糖乳杆菌(Lactobacillus rhamnosus),枯草芽孢杆菌(Bacillus subtilis),腐生葡萄球菌(Staphylococcus saprophyticus),木糖葡萄球菌(Staphylococcus xylosus),戊糖片球菌(Pediococcus pentosaceus),植物乳杆菌(Lactobacillus plantarum)。为发酵肉制品的工业化生产和安全性提供了良好的理论依据。
In order to obtain good amine oxidase productive starters for meat product,amine oxidase productive strains were isolated from 44 samples in different areas of the smoked horse intestine.Productive amine bacteria were eliminated by double color method in monolayer culture.The oxidase test,bacteriostasis experiment and the resistance to salt and nitrite test were conducted for isolated strains.The same strains were eliminated by DGGE and the ability of oxidizing biogenic amine was verified by HPLC.Finally,the DNA was extracted from the screened strains,and V6-V8 segment of 16s rDNA were sequenced after PCR amplification.The results showed that six strains productive amine oxidase bacteria were screened out,with salt tolerance above 6%,nitrite tolerance above 0.15 g/kg,and had good inhibitory effect to Escherichia coli,Listeria monocytogenes,and Staphylococcus aureus,and they had strong oxidizing ability of biogenic amines.The strains were Lactobacillus rhamnosus,Bacillus subtilis,Staphylococcus saprophyticus,Staphylococcus xylosus,Pediococcus pentosaceu and Lactobacillus plantarums.This research provides a good theoretical basis for the industrial production and security of fermented meat.