以Indole-3-acetic acid為指標生產具有抗發炎活性的玉米黑穗菌發酵液

Production of the fermentation broth of Ustilago maydis possessing anti-inflammation function using indole-3-acetic acid as marker

詹宗達(Tsung-Ta Chan)

指導教授 ： 蔣丙煌

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2012年)

https://doi.org/10.6342/NTU.2012.10031

摘要

玉米黑穗菌是一種寄生在玉米上的寄生菌，在玉米上生成膨大的腫瘤組織，在墨西哥當地被視為一種傳統美食。研究指出，此菌會代謝色胺酸，生成如indole-3-acetic acid(IAA)以及pityriarubin類化合物，其分別在動物實驗和細胞實驗上證實具有抗發炎效果。因此，本研究即期望利用此菌，以IAA為指標，使用攪拌式發酵槽生產具有抗發炎活性的玉米黑穗菌發酵液。經過一系列的最適化研究，結果發現在培養溫度24 ℃、起始pH 4、培養基組成為含有1 g/L色胺酸的PDB培養基額外添加20 g/L葡萄糖、扇葉攪拌速率900 rpm、通氣量0.5 vvm的條件下，經過32小時的發酵時間，可生產出含有30.8±4.5 μg/ml IAA (HPLC定量)的玉米黑穗菌發酵液，並以RAW264.7細胞模式證明此發酵液在10%的添加量下，其抑制發炎因子NO的比例達81.0±0.3%。

關鍵字

玉米黑穗菌 ； Indole-3-acetic acid ； IAA ； 抗發炎 ； 發酵

並列摘要

Ustilago maydis is a pathogen infecting on corn and induces tumor tissue formation. Althoug it is a pathogen, it is seen as a traditional cusine in Mexico. It can metabolize trytophan into indole-3-acetic acid (IAA) and pityriarubin with anti-inflammation bioactivity. Therefore, the goal of this research was to produce fermentation broth of Ustilago maydis possessing anti-inflammation function using indole-3-acetic acid as marker in a 5 L stirred tank fermentor. After optimization process, it was found that the fermentation process would produce 30.8±4.5 μg/ml IAA in the broth using PDB medium supplemented with 1g/L tryptophan and 20 g/L glucose with initial pH at 4. The optimal fermentation conditions in the stirred tank bioreactor are: agitation rate at 900 rpm; aeration rate at 0.5 vvm, and 24℃ for 32 h. Moreover, the broth (10%) can inhibit nitrite production up to 81.0±0.3% in RAW264.7 inflammation cell model.

並列關鍵字

Ustilago maydis ； Indole-3-acetic acid ； IAA ； anti-inflammation function ； fermentation

參考文獻

6. Banuett, F., Pathogenic development in Ustilago maydis: a progression of morphological transitions that results in tumor formation and teliospore production. Mycol. Ser. 2002, 15, 349-398.

9. Brandl, M. T.; Lindow, S. E., Environmental signals modulate the expression of an indole-3-acetic acid biosynthetic gene in Erwinia herbicola. Mol. Plant. Microbe Interact. 1997, 10, 499-505.

10. Broek, A. V.; Gysegom, P.; Ona, O.; Hendrickx, N.; Prinsen, E.; Van Impe, J.; Vanderleyden, J., Transcriptional analysis of the Azospirillum brasilense indole-3-pyruvate decarboxylase gene and identification of a cis-acting sequence involved in auxin responsive expression. Mol. Plant. Microbe Interact. 2005, 18, 311-323.

11. Budde, A. D.; Leong, S. A., Characterization of siderophores from Ustilago maydis. Mycopathologia 1989, 108, 125-133.

12. Chung, K. R.; Tzeng, D. D., Biosynthesis of indole-3-acetic acid by the gall-inducing fungus Ustilago esculenta. J. Biol. Sci. 2004, 4, 744-750.