新疆熏马肠中产氨基酸脱羧酶优势细菌的分离及鉴定

Separation and Identification of Dominant Bacterial Strains Producing Amino Acid Decarboxylase in Xinjiang Smoked Horsemeat Sausage

本研究从新疆熏马肠中分离鉴定得到32株产氨基酸脱羧酶的优势细菌,并确定了各株菌的产胺能力。试验从新疆多产熏马肠的四个地区采集44个熏马肠样品,利用双层培养基显色法初步分离其中的产胺优势菌。再采用PCR-DGGE技术,剔除重复菌株并通过同源性比对,确认32株产胺菌。最后利用高效液相色谱检测菌液产物,验证其产生物胺的能力。此次研究中,得到的菌株主要高产腐胺、色胺、组胺及苯乙胺,其中4号菌株(气味沙雷氏菌,Serratia odorifera)产生物胺总量(平均值)最高985.14μg/mL,且组胺生成量最高7259.64μg/mL;17号菌株(阴沟肠杆菌,Enterobacter cloacae)产生物胺总量(平均值)次高737.78μg/mL,且色胺生成量最高4507.81μg/mL;11号菌株(克雷伯氏菌,Klebsiella sp.)产腐胺量最高2550.54μg/mL;8号菌株(木糖葡萄球菌,Staphylococcus xylosus)产苯乙胺量最高3831.50μg/mL。

In this study,32 dominant bacterial strains producingamino acid decarboxylase were isolated from 44 samples of Xinjiang smoked horsemeat sausage,collected from four districts in Xinjiang where producing smoked horsemeat sausage.The ability of the isolates to producebiogenic amines was determined and dominant biogenic amine-producing strains were distiguished by a chromogenic reaction in double-layered media.Subsequently,polymerase chain reaction/denaturing gradient gel electrophoresis（PCR-DGGE） was used to reject repetitive strains and 32 biogenic amine-producing strains were identified viahomology comparison.Finally,high-performance liquid chromatography（HPLC） was used to examine bacterial broth to confirm the production of biogenic amine.The strains thus obtained had strong ability to produce putrescine,tryptamine,histamine,and phenylethylamine.Strain number 4（Serratia odorifera） showed the highest average production of total biogenic amines（985.14 μg/mL） and the highest production of histamine（7259.64 μg/mL）.Strain number 17（Enterobacter cloacae） showed the second-highest production of total biogenic amines（737.78 μg/mL） and the highest production of tryptamine（4507.81 μg/mL）.Strain number 11（Klebsiella sp.） showed the highest production of putrescine（2550.54 μg/mL）,whereas Strain number 8（Staphylococcus xylosus） showed the highest production of phenylethylamine（3831.50 μg/mL）.