摘要
为开发绿色野生药用大型真菌源饲料抗菌添加剂，本研究采用杯碟法筛选活性菌株和一剂量法测定效价，通过单因素试验法、Plackett-Burman筛选、最陡爬坡、中心组合试验、响应面分析及正交试验优化抗菌物质的发酵生产工艺，获得了一株对病原菌具有高效、广谱抗性的野生冬菇0612-9，其5倍浓缩发酵液对金黄色葡萄球菌、大肠杆菌和白色念珠菌的抑菌圈直径分别为23.26 mm、16.12 mm和19.70 mm，与盐酸四环素比，抗金黄色葡萄球菌的相对效价为1806.34 U/mL。0612-9菌株发酵生产抗菌物质的最佳工艺为：土豆270.53 g/L，麸皮7.429 g/L，葡萄糖30 g/L，MgSO4·7H2O 0.4551 g/L，KH2PO4 2 g/L，培养基初始pH值为4.0，发酵周期11 d，接种量5 mL/100 mL，转速180 r/min，摇床温度27 ℃，装液量100 mL/250 mL。该胞外抗菌物质在低于60 ℃、pH1.0～6.0抑菌活性稳定，对紫外照射不敏感，乙酸乙酯为抗菌物质的最佳提取剂。
To develop green feed antibacterial additives originated from wild edible and medicinal fungi, a total of 65 fungi were screened by cup plate method, the potency were tested by a dose method and the process for the production of antimicrobial substances were optimized by single factor experiment, the Plackett-Burman screening, steepest ascent, central composite design, response surface analysis and orthogonal test. The ＂0612-9＂, a kind of wild mushroom produced the substances which possessed broad-spectrum antibacterial activity. The antibacterial circle diameter of 5 times concentrated fermentation broth on Staphylococcus aureus, Escherichia coli and Monila albicans were 23.26 mm, 16.12 mm and 19.70 mm, the relative potency against S.aureus was 1.806.34 U/mL. The optimal fermentation parameters were 270.53 g/L of potatoe, 7.429 g/L of wheat bran, 30 g/L of glucose, 0.4551 g/L of MgSO4·7H2O and 2 g/L of KH2PO4. The initial pH value of the medium was 4.0, the fermentation period of 11day, packing in the volume of 100 mL/250 mL, inoculating at 5 mL∶100 mL, rotating at speed of 180 r/min and incubating at 27 ℃ respectively. The extracellular antibacterial substances could remain active less than 60 ℃, pH1.0-6.0.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2014年第3期208-213,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
南昌市生物资源保护和利用重点实验室项目(2012-sys-04)
关键词
野生冬菇
发酵工艺
响应面法
抗菌物质
Flammulina ssp
fermentation process
response surface methodology
antibacterial material
