Production of Anti-Fungal Agent by Microorganisms: Screening Microorganism and Optimal Condition for---(III)

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Project title

Code/計畫編號

NSC89-2313-B019-073

Translated Name/計畫中文名

以微生物生產真菌抑制劑：生產菌株的篩選與最適生產條件的探討(III)

Project Coordinator/計畫主持人

Guo-Jane Tsai

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=553417

Year

2000

Start date/計畫起

01-08-2000

Expected Completion/計畫迄

01-07-2001

Bugetid/研究經費

549千元

ResearchField/研究領域

食品科技(農)

Candida albicans為人體之機緣性致病酵母菌，本研究以Candida albicans為作用目標菌，從附著在海洋固體表面及土壤中，篩選得最具潛力之真菌抑制劑生產菌株，經鑑定為Trichoderma parceramosum Bissett No. 017，分別探討碳源、氮源、鹽類及培養溫度與時間對其生產之真菌抑制劑之影響，並以此最適條件來生產真菌抑制劑。當培養液以Glucose為碳源、以Beef extract為氮源時最有利於此菌生產真菌抑制劑。培養液添加0.2% NaCl，可增加真菌抑制劑之產生。在1% Glucose，1.1% Beef extract及0.2% NaCl，起始pH值為pH8之培養液中，此菌之最適生長溫度為25-30℃，培養7天後，培養液中真菌抑制劑活性達最高。濾液中的真菌抑制劑對高溫之耐受性高，在100℃加熱10分鐘仍保有67%活性；pH對此粗抑制劑安定性無明顯影響；其不受Papain及Amylase之影響，但Lipase會降低其活性；分子量介於3,000~10,000 Da；冷藏或冷凍六週活性不變。除了Candida albicans外，此真菌抑制劑對Candida albicans CCRC 21538、Filobasidiella neoformans var. neoformans CCRC 20533及Trichosporon beigelii CCRC 21675等三株病原酵母菌及Aspergillus niger CCRC 30201、Aspergillus parasiticus CCRC 30117、Penicillium minioluteum CCRC 32639及Penicillium minioluteum CCRC 32645等四株病原黴菌均具極強的抑菌活性。經超過濾濃縮及Gel fitration處理後，可得到純化之真菌抑制劑，其耐熱性略低於未純化狀態，對酵素的耐受性則較未純化者高。對Candida albicans CCRC 20513、Candida albicans CCRC 21538、Candida glabrata CCRC 20586、Candida krusei CCRC 21543亦有極強之抑菌活性。動物感染試驗顯示口服此真菌抑制劑可有效提高小白鼠對抗Candida albicans感染之能力，其效果甚至比目前普遍使用之真菌藥物Fluconazole還好。 Candida albicans is an opportunistic pathogenic yeast of humans. Among thousands of fungal isolates from various substrate exposed to seawater and from soil, an isolate identified as Trichoderma parceramosum Bissett No. 017 was screened out for the production of anti-Candida albicans agent. The effects of carbon and nitrigen source, salts, pH value and cultivation temperature on the production of this antifungal agent were investigated. Glucose and beef extract were the best carbon source and nitrogen source in the medium for production of antifungal agent. The addition of 0.2 % NaCl stimulated the production of this antifungal agent. The optimal production temperature was in the range of 25-30.degree.C in the medium which was composed of 1% glucose, 1.1% beef extract and 0.2 % NaCl with initial pH value of pH 8. The crude antifungal agent was thermal stable, with residual activity being 67% after heating at 100.degree.C for 10 min. It was stable to pH variation and to freezing and refrigeration storage for 6 weeks. Its molecular weight was about 3,000~10,000 Da, and this antifungal agent was resistant to papain and amylase. Except Candida albicans, this antifungal agent showed strong activity against various pathogenic fungi including Candida albicans CCRC 21538, Filobasidiella neoformans var. neoformans CCRC 20533, Trichosporon beigelii CCRC 21675, Aspergillus niger CCRC 30201, Aspergillus parasiticus CCRC 30117, Penicillium minioluteum CCRC 32639, and Penicillium minioluteum CCRC 32645. The antifungal agent can be purified by amicon membrane sepration first, followed by gel fitration chromatography. The purified antifungal agent was less thermal stable than crude one, but higher resistant to various digestive enzyme. This purified antifungal agent also showed strong activity against Candida albicans CCRC 20513, Candida albicans CCRC 21538, Candida glabrata CCRC 20586, and Candida krusei CCRC 21543. Oral digestion of this antifungal agent can increase the resistance of mice against the infection of Candida albicans. The survival rate for antifungal agent obtained was even higher than that of fluconazol, a commercial antifungal drug widely used in clinical treatment.

Keyword(s)

白念珠菌
真菌抑制劑
木黴菌
致病性酵母菌
抗菌圖譜
生產條件
Candida albicans
Antifungal agent
Trichoderma
Pathogenic yeast
Antimicrobial spectrum
Production condition

DSpace CRIS

Production of Anti-Fungal Agent by Microorganisms: Screening Microorganism and Optimal Condition for---(III)

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