摘要
目的研究Rhizobium sp.N613胞外多糖(REPS)的分离纯化条件和抑瘤活性。方法用酸化、加热、凝聚剂和絮凝剂进行发酵液预处理实验,经正交试验优化REPS醇沉制取条件。用凝胶柱色谱,紫外光谱扫描检测经Sevag法脱蛋白质所得REPS的纯度。采用动物移植性实体瘤的瘤重实验法研究REPS对小鼠S180肉瘤的抑瘤作用,并对荷瘤小鼠眼眶取血进行白细胞及淋巴细胞计数。结果发酵液预处理参数为:盐酸调节pH5.0,70℃加热20min,依次加入5g·L-1Al2(SO4)3.18H2O和0.2g·L-1海藻酸钠。优化的醇沉制取条件为:调节溶液pH7.0,乙醇浓度65%,醇沉时间8h。REPS经检测为均一多糖。抑瘤实验结果表明,REPS各剂量组对小鼠S180肉瘤均有抑制作用,其中REPS纯多糖剂量为10mg·kg-1的抑瘤率达44.17%。与荷瘤对照组相比,REPS能够显著促进白细胞和淋巴细胞的增生。结论初步摸索到了发酵液预处理操作的技术参数与醇沉制取的条件。所得REPS对小鼠S180肉瘤有显著的抑制作用。
Abstract
OBJECTIVE To study the isolation and anti-tumor activity of exopolysaccharide(REPS) from Rhizobium sp.N613.METHODS The fermentation broth was pretreated by acidification,heating,adding coagulant and flocculant.The ethanol precipitation conditions of REPS was optimized by an orthogonal design.REPS was deproteined by Sevag method,and homogeneity was determined by gel column chromatography and UV spectrum.Transplanted tumor models of sarcoma 180 in mice were used to evaluate the anti-tumor effect.The white blood cell and lymphocyte of blood samples from S180 bearing mice were counted.RESULTS The process of fermentation broth pretreatment adjusted pH value to 5.0 with HCl at 70 ℃ for 20 min,with the addition of 5 g·L-1 Al2(SO4)3·18H2O and 0.2 g·L-1 sodium alginate.The optimized conditions of REPS ethanol precipitation were as follows:pH value 7.0,65% ethanol and precipitation time for 8 h.REPS was homogeneous polysaccharide.The result of anti-tumor activity showed that inhibition rate was 44.17%.Compared with the S180 bearing group,REPS significantly promoted the proliferation of white blood cell and lymphocyte.CONCLUSION The parameters of fermentation broth pretreatment and the conditions of REPS ethanol precipitation were obtained.REPS inhibited significantly the growth of S180.
关键词
根瘤菌 /
胞外多糖 /
分离 /
抑瘤
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Key words
Rhizobium sp. /
exopolysaccharide /
isolation /
anti-tumor
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程红兵;韩勇;任盛;梁丽绒;赵良启.
Rhizobium sp.N613胞外多糖分离纯化及抗S180肉瘤的研究[J]. 中国药学杂志, 2008, 43(04): 268-271
CHENG Hong-ing;HN Yong;REN Sheng;LING Li-rong;ZHO Ling-qi.
Studies on Isolation and Anti-S180 Activity of Exopolysaccharide from Rhizobium sp.N613 [J]. Chinese Pharmaceutical Journal, 2008, 43(04): 268-271
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参考文献
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脚注
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基金
山西省自然科学基金资助项目(2006011073);山西高校科技研究开发资助项目(20051203)
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