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  • 學位論文

菰黑穗菌發酵液成分分析之探討 :( 1 )以反應曲面法生產stigmastane-3,6-dione、( 2 )以酒粕液刺激生產成分A-1之結構與生化活性的研究

The Production Of Stigmastane-3,6-dione From Ustilago Esculenta Using Response Surface Methodology, And The Evaluation Of Bioactivity And Structural Analysis Of Composition A-1 Stimulated By Distiller Grain Liquid

指導教授 : 吳立真
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摘要


Stigmastane-3,6-dione被發現有許多生物活性,但取得方式耗費時間、 金錢,而且獲得的量也少,所以想利用菰黑穗菌去大量生產stigmastane-3,6-dione,實驗第一部分是使用反應曲面法來找尋菰黑穗菌大量生產stigmastane-3,6-dione的條件。菰黑穗菌已被報導可以抗骨質疏鬆以及其抗骨質疏鬆物質也被確定是stigmastane-3,6-dione。其中改變因子有培養溫度、培養轉速及起始pH值,結果得到在溫度27.93℃、轉速119.41 rpm及起始pH值5.01時,stigmastane-3,6-dione可達到最大濃度53.4 mM。 本來想使用酒粕液去刺激菰黑穗菌去大量生產stigmastane-3,6-dione,但是產量不但沒有提升,而且還下降,但是意外發現培養液變成黏稠狀,推測有大量的成分A產生,故本實驗第二部分使用酒粕液去刺激菰黑穗菌生產大量的成分A-1,發現酒粕液 : PDB培養基= 15 : 85 時,成分A-1有最大的產率,將此成分A-1進行化學結構及生物活性的分析,發現組成單糖可能是葡萄糖及甘露糖,而鍵結是以1-6鍵結最多、1-3鍵結第三多,而抗發炎實驗則是添加到100μg/ml的成分A-1開始有效果且添加越多效果越強,但EC50的結果是超過800μg/ml,可能是結構組成差異,造成抗發炎效果不明顯。

並列摘要


Stigmastane-3 ,6-dione have been found in many biological activities, but it takes a lot of time and money to acquire just a small amount of it. In this study, we have utilized Ustilago Esculenta to produce a numerous amount of stigmastane-3,6-dione. The first part of the experiment was to use what we called the Response Surface Methodology for finding the optimal condition in producing stigmastane-3,6-dione with the use of Ustilago Esculenta. Ustilago Esculenta has been reported to slow down the osteoporotic process, which has been believed to be induced by stigmastane-3, 6-dione. Also, there are many factors that come into play in producing stigmastane-3 ,6-dione such as the incubation temperature, rotational speed of the culture and the initial pH values. Under these conditions, the maximal concentration of stigmastane-3 ,6-dione has been reached and it was 53.4 mM, at the temperature of 27.93℃, the rotational speed of 119.41 rpm and the initial pH of 5.01. Originally, it was our intention to use distilled grain liquid to stimulate Ustilago Esculenta for the growth of stigmastane-3 ,6-dione; as a result, the yield has become significantly decreased. However, from this experiment, we have found out the culture medium has become viscous, assuming that there was a tremendous production of composition A-1 because of that. Therefore, the second part of the experiment was to use distiller grain liquid to stimulate Ustilago Esculenta to produce large amount of composition A-1. Found in distillers grains liquid: PDB medium = 15 : 85., the composition A-1 have the greatest yield. As a result, the analysis of the chemical structures and biological activities of composition A-1 has been done, the composition of monosaccharides could possibly be glucose and mannose with maximum amount of 1-6 bonding, followed by1-3 bonding. The anti-inflammatory study has a positive effect when added with 100μg/ml composition A-1, and a stronger effect with a larger dose. But, the EC50 result was over 800μg/ml, due to different in composition and structure, resulting the anti-inflammatory effect was not obvious.

參考文獻


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