高产D-甘露醇虫草菌种分离纯化的研究

本试验系用生物工程技术分离纯化虫草菌种的研究。先用组织分离法从野生虫草整体组织上分离出虫草菌种 ,再用平板稀释法将菌种纯化 ,获得一株高产D -甘露醇 (虫草酸 )的优良菌种。该菌种被鉴定为蝙蝠蛾拟青霉。使用该菌种经一级种子培养和发酵培养 ,获得含D -甘露醇 9 9% (野生虫草约 7% )、总氮 5 1% (野生虫草约 4% )的虫草菌丝体。原种斜面培养基组成 (% ) :琼脂 2、葡萄糖 2、蛋白胨 0 2、磷酸二氢钾 0 1、硫酸镁 0 0 3、麦麸 5。种子培养基组成 (% ) ;葡萄糖 2、蛋白胨 0 5、磷酸二氢钾 0 1、硫酸镁 0 0 3、麦麸 5。原种培养温度 2 3- 2 4℃、培养时间 4- 5天 ;平均稀释纯化所用稀释菌液为 10 -1、 10 -2 、 10 -3 、 10 -4 、 10 -5、 10 -6,挑菌培养温度分别为 2 3- 2 4℃、 2 8℃、 37℃。种子培养条件 :一级种子培养 :摇床转速 180r/min ,培养温度 2 4- 2 5℃ ,时间 2 - 3天 ;二级种子培养 ;摇床转速 90r/min ,培养温度 2 4- 2 5℃ ,时间 2天。发酵培养条件 ;接种量 5 - 10 % ,温度 2 4- 2 6℃ ,搅拌速度 15 0 - 180r/min ,通气培养 ,时间 5 - 6天 ,PH4 5 ,残糖量降至 0 3%。

The Study of Separating and Purifying of Cordyceps Sinensis of High Produce D- Mannitol

It is a study of separating and purifying the cordyceps sinensis agaric by the biological enginering. Firstly, separate the agaric of cordyceps sinensis from the whole organizations of wild cordyceps sinensis by the way of fissue organic separation. Secondly, purify the agaric vaccine with the plate dilution method. Thus, we can gain an excellent high production agaric vaccine, D-mannitol. Ferment and nourish the agaric vaccine in the-one-standing conditions, and we can have the myceda including 9.9% of the D-mannitol and 5.1% of the total nitrogen (the quantities of which in the wild cordyceps sinensis are 7% and 4%). The compositions of the original vaccine's inclined plane growth medirm:agar 2, grape glucose 2, protein hydrolysate 0.2, potassium dihyolrofen phosphate 0.1, magnesium sulfate 0.03, wheat bran 5. The compositions of the spore's inclined plane growth medirm:grape glucose 2, protein hydrolysate 0.5, potassium dihyolrofen phosphate 0.1. magnesium sulfate 0.03. Wheat bran 5. The temperature of the original vaccine nourishing:23-24℃. The time of the original vaccine nourishing:4-5 days. The diluting liquid by the plate dilution method:10 -1 ,10 -2 ,10 -3 ,10 -4 ,10 -5 ,10 -6m . The temperature of nourishing pick mash room: 23-24℃,28℃,37℃. The condition of the spore nourishing: The-one-standing spore nourishing:the stirring speed of vibrator:180r/min, nourishing temperature:24-25℃, time:2-3 days. The-two-standing spore nourishing:the stirring speed of vibrator:90r/min, nourishing temperature:24-25℃, time:2 days. The condition of the fermentative nourishing:the stirring speed of vibrator:150-180r/min, rentinate growth:5-6 days, pH:4.5. The degree of the surplus sugar number's reduce:0.3%.