摘要
以摇床振荡培养猪囊虫病疫苗用ＳＯＡ工程菌，确定了其最佳培养条件：培养基为含０．２％葡萄糖的ＬＢ磷酸盐复合培养基（ｐＨ７．４），诱导剂为１％乳糖，培养温度为３７℃。以此为基础进行工艺放大，研究了ＳＯＡ工程菌在７０Ｌ国产发酵罐中的发酵培养工艺，确定了接种量、ｐＨ、气压、溶解氧、温度等参数。试验结果显示，种子罐细菌量为ＯＤ６００＝４．０时，接种到发酵罐的菌体收获量最高；罐体气压在０．０５～０．２５ＭＰａ之间变动对工程菌产量、活菌数及表达量等均无明显影响；整个发酵过程中，发酵条件比较稳定，ｐＨ为７．４～７．０，溶解氧保持在１００以上，温度３６～３８℃，发酵时间８～１０ｈ。发酵结束，发酵液ＯＤ６００高达１１．２，活菌数为１０３亿／ｍＬ，表达量为１６．２％，重组质粒稳定。
Cultivation condition of recombinant Escherichia coli (SOA) which would be expected to produce the vaccine against swine cysticercosis were first determined in a rotary shaker. The best cultivation conditions were set as follows: culture medium was a complex LB medium with 0.2% glucose and phosphate salt of pH7.4; its inducer being 1% lactose and the culture temperature of 37℃. This culture process could be scaled up in a 70 L made in China fermentor based on the above condition, then determined inoculum volume, pH, gas pressure in fermentor, dissolved oxygen and temperature. When OD 600 of seed fermentor was 4.0, the germ weight of the recombinant E. coli yielded best. The gas pressure between 0.05 0.25 Mpa didn′t influenced the total germ weight, the number of vial bacteria and gene expression. The fermentation conditions among all the fermenting process were relatively stable, with the pH of 7.4 7.0, the dissolved oxygen to be kept above 100, its temperature of 36 38℃, the total fermenting time of 8 10 h. At the end, the optical density (OD 600 ) of the fermented liquid reached at 11.2, the colony forming unit (cfu/mL) measured up to 103×10 8, the recombinant plasmid remained stable.
出处
《中国兽医学报》
CAS
CSCD
北大核心
1999年第2期161-164,共4页
Chinese Journal of Veterinary Science
关键词
猪
囊虫病
基因工程疫苗
SOA工程菌
培养条件
swine cysticercosis
genic recombinant vaccine
recombinant Escherichia coli
cultivation condition
fermentation condition
