【作者】 王晓玲；

【导师】 周国英；

【作者基本信息】 中南林学院 ， 森林保护学， 2005， 硕士

【摘要】 本论文主要研究药用真菌紫芝的深层发酵工艺,探讨虫类药物九香虫对紫芝发酵生产抗癌活性物质紫芝多糖和三萜类物质的影响,运用正交设计对紫芝—九香虫发酵生产胞内多糖和胞外三萜类物质进行优化,采用MTT法对所得产物的体外抗肿瘤活性进行了研究。研究结果如下: 1、适宜紫芝液体发酵的粮食型培养基为:米糠6g/L,玉米粉20g/L,黄豆粉12g/L,蔗糖20g/L。此培养基培养的紫芝生物量可达15.12g/L。 2、在紫芝培养基中添加适量的九香虫,对紫芝的生长影响不大,但能显著提高紫芝的胞内多糖和胞外三萜类物质的产量。在紫芝培养基中添加5%的九香虫,紫芝胞内多糖的产量显著提高,由原来的75.17.mg/g菌体提高到105.01mg/g菌体;胞外多糖也有所提高,由原来的24.58mg/100mL提高到28.09mg/100mL。胞外三萜类物质的含量由原来的132.6mg/L提高到157.1mg/L。但当九香虫的添加量增大到8%时,菌体生长明显受到抑制,多糖和三萜类物质的产量也明显低于对照。 3、九香虫提高紫芝活性物质产量的有效成分的研究表明,九香虫促进紫芝胞内多糖合成的有效成分主要是九香虫水提取物;提高胞外三萜类物质的产量的有效成分主要是九香虫乙醇提取物:促进胞外多糖分泌的有效成分主要是九香虫乙醚提取物。 4、紫芝活性成分的发酵优化,即九香虫添加量、添加时间和总培养时间的优化结果表明,以胞内多糖产量为指标时最佳添加量为5%,添加时间为发酵起始1天后加入,培养时间为6天,胞内多糖产量可达109.28mg/g,比优化前的105.01mg/g提高了4.27 mg/g:以胞外三萜类物质产量为指标的添加量为5.5%,添加时间为发酵起始1天后加入,总培养时间为7.5天,胞外三萜类物质产量为159.8mg/L,比优化前的157.1 mg/L提高了2.7 mg/L。 5、紫芝三萜类物质的体外抗肿瘤实验表明,在300μg/mL剂量下,紫芝-九香虫发酵组对人肠癌细胞SW620的抑制率为46.01%,紫芝单独发酵组的抑制率为45.89%;在100μg/mL剂量下,紫芝-九香虫发酵组的抑制率为36.76%,紫芝单独发酵组的抑制率为36.84%。两组样品对人正常肝细胞株L一02的抑制作用都不显著。更多还原

【Abstract】 It was studied in the dissertation, the optimum media for submerged fermentation of Ganoderma sinense, effect of Aspongopus chinensis Dallas on content of anti-cancer active substances polysaccharides and triterpenenoids from Ganoderma sinense by submerged fermentation, optimizing fermentation condition of Ganoderma sinense for active component of polysaccharides and triterpenenoids, and anti-tumor action of triterpenenoids from Ganoderma sinense on SW620.1、 The results showed that the optimum media for submerged fermentation of Ganoderma sinense were made up of the following components (in grams per litre): rice bran 6, corn flour 20, bean flour 12, sucrose 20. The biomass reached 15.12 g/L in this media.2、 A properly amount of Aspongopus chinensis Dallas in media of G. sinensehas little effect on growth of G. sinense, but markedly improved the output of IPS(intracellular polysaccharides) and ETD (exo- triterpenenoids). The output of IPSincreased from 75.17 mg/g to 105.01mg/g, while the output of EPS(Exo-polysaccharides) only increased from 24.58 mg/100mLto 28.09mg/100mL.At the same time, the output of ETD has an improvement of 24.5 mg/L when added5% Aspongopus chinensis Dallas in the media.3、 The effective component of Aspongopus chinensis Dallas for improving the output of IPS of G. sinense may be aqueous extract of Aspongopus chinensis Dallas; The effective component improving the output of ETD may be alcohol extract of Aspongopus chinensis Dallas. Additionally, the ether extract of Aspongopus chinensis Dallas could also improve the secretion of EPS.4、 The optimizing fermentation conditions: when IPS is main target , the optimum addition amount of Aspongopus chinensis was 5g/L, added time was 1 d later from initial fermentation, total culture time was 6 d; the output of IPS reached 108.28 mg/g. while ETD is main target, the amount was 5.5g/L, time was 1 d later too, total culture time was 7.5 d; the output of ETD reached 159.5 mg/L.5、 The fermented substances in the study were tested their extracellular action on human tumor cell by MTT method. The ETD from fermentation both G. sinense and Aspongopus chinensis could markedly inhibit the proliferation of SW620 human更多还原