摘要
以细胞碎片得率为指标,在单因素试验的基础上采用二次回归正交旋转组合设计进行红色诺卡氏菌菌体破壁工艺优化,以期提高破壁效率。根据回归模型进行重要性效应分析,通过频率分析法获得超声破壁最佳工艺参数:固液比1∶2.5、超声功率744 W、每次超声时间3 min、全程超声时间51 min,此工艺参数下,细胞碎片得率可达到约28.3%,相比原工艺上升了62.4%。在扫描电镜下观察菌体破壁前后形态,发现易破壁菌体为长杆状而难破壁菌体趋近于圆球状,优化后工艺参数所得的碎片较小;通过荷瘤小鼠体内抑瘤实验发现小碎片对S-180瘤株的抑制作用约有22.1%提升(抑瘤率分别为40.7%和49.4%),优化积极意义明显。
Using the cell debris yield as target and based on the single factor test,the quadratic regression orthogonal rotary combi- nation method was designed to optimize cell wall disruption of Nocadia rubra, which was in hope of improving wall-cracking efficien- cy. According to the regression model analysis of the main effection, the optimum parameters of ultrasonic were determined by frequency analytic approach：solid to liquid ratio 1：2.5 （g/mL） , ultrasonic power 744 W, each ultrasonic time 3 rain and the total ultrasonic time 51 min. Under such conditions ,the cell wall fragments yield reached 28.3% ,which was 62.4% higher than the origi- nal process. Using scanning electron microscope, the mycelial morphology of the before and after wall-broken was observed. It Was found that the easy form of wall cracking thalli was trend of long rod-shaped while the hardly wall cracking thalli was Spherical. Under the optimization of process parameters, the fragmentation was relatively small . Through the tumor-bearing mice experiment in vivo,It was found that small pieces of inhibitory effect on S-180 cell line increased about 22. 1% （ The tumor inhibition rate was 40.7% and 49.4% ） ,which was more suitable for the follow-up process.
出处
《药物生物技术》
CAS
2013年第6期522-527,共6页
Pharmaceutical Biotechnology
基金
福建省科技2013年度区域科技重大项目(No.2013Y3001)
关键词
红色诺卡氏菌
超声波
二次回归正交旋转组合设计
扫描电镜
最优化
Nocadia rubra, Ultrasonic, Quadratic orthogonal rotation combination design, Scanning electron microscope, Optimum
