蜜环菌YN3862菌株的高密度发酵工艺

High-density Fermentation Technology of Armillaria mellea YN3862 Strain

以蜜环菌YN3862菌株为试材,采用固体培养和液体深层发酵相结合的方法,研究蜜环菌的最优母种培养基和最佳发酵条件,以期达到高密度培养蜜环菌目的。结果表明:该菌株最适母种培养基成分为木屑100 g·L^-1,麦麸100 g·L^-1,葡萄糖20 g·L^-1,琼脂15 g·L^-1,牛肉膏5 g·L^-1;最佳液体培养基成分为葡萄糖1%,蔗糖2%,蚕蛹粉3%,MgSO4·7H2O 0.075%,KH2PO40.15%,乙醇1%;液体深层发酵采用200 L发酵罐,装入100 L液体培养基,以5%接种量接入匀质后的液体菌种,发酵温度25℃,溶解氧含量(DO)为55%左右,在发酵的0~24、24~36、36~48、48~144 h,转速分别设置为60、80、120、150 r·min^-1,在发酵时间为120 h时,生物量(干质量)最高,达到2.5 g·(100mL)^-1。因此,采用上述培养基和液体深层发酵工艺,产出的生物量大,菌丝球活力好,实现了高密度培养。

Armillaria mellea YN3862 strain was used as test material,a combination of solid culture and deep liquid fermentation was used to study the optimal mother culture medium and fermentation conditions of A.mellea,in order to achieve the purpose of high-density culture of A.mellea.The results showed that the best culture medium for A.mellea includes sawdust 100 g·L^-1,wheat bran 100 g·L^-1,glucose 20 g·L^-1,agar 15 g·L^-1 and beef extract 5 g·L^-1.The optimum liquid medium composition was 1%glucose,2%sucrose,3%silkworm pupa powder,0.075%magnesium sulfate heptahydrate,0.15%potassium dihydrogen phosphate and 1%ethanol.Deep liquid fermentation was conducted in a 200 L fermentation tank and filled with 100 L liquid culture medium,the liquid strain was inoculated with 5%inoculation amount,the fermentation temperature was 25℃and the dissolved oxygen was about 55%.At 0-24,24-36,36-48,48-144 hours of fermentation,the rotational speeds were set as 60,80,120,150 r·min^-1,respectively.The optimal fermentation time was 120 hours.At this time,the mycelium quantity could reach 2.5 g·(100 mL)^-1.In conclusion,by using the above medium and deep liquid fermentation technology,the biomass produced was large,and the produced mycelium pellet had good vitality and realized high-density culture.