微生物學通報 OCT 20,2010,37(10):1475～1480
浙貝母內生真菌菌株FTJZZJ09及其抑菌活性成分
田峰 何仁發(fā) 李明月 王娟 葉波平*
(中國藥科大學生命科學與技術學院 江蘇 南京 210009)
摘 要: 根據菌株的形態(tài)學特征和ITS序列分析結果, 將一株來源于浙貝母新鮮鱗莖的內生真菌菌株FTJZZJ09鑒定為產黃青霉(Penicillium chrysogenum), 其在優(yōu)化后的改良查氏培養(yǎng)基(3 g/L麥芽糖, 3 g/L蛋白胨, 0.1 g/L K2HPO4, 0.05 g/L KCl, 0.3 g/L NaNO3, 0.05 g/L MgSO4·7H2O, 0.001 g/L FeSO4·7H2O, pH 6.5)中28°C、160 r/min振搖培養(yǎng)7 d, 獲得的發(fā)酵液具有抑菌活性,。利用正相硅膠,、Sephadex LH-20柱層析以及重結晶等方法, 從發(fā)酵液乙酸乙酯提取物中分離到3個具有抑菌活性的化合物, 利用質譜和核磁共振等技術將其分別鑒定為環(huán)(脯氨酸-甘氨酸)、環(huán)(脯氨酸-纈氨酸)和2-乙?；?4 (3H)喹唑酮, 它們對枯草芽孢桿菌的生長均具有較為明顯的抑制作用, MIC分別為0.8,、0.8和0.4 g/L, 但對革蘭氏陰性菌的生長無明顯的影響。
關鍵詞: 浙貝母, 內生真菌, 產黃青霉, 二酮哌嗪類, 喹唑酮類
The Endophytic Fungus Strain FTJZZJ09 Isolated from the Bulbs of Fritillaria thunbergii and Its
Antibacterial Metabolites
TIAN Feng HE Ren-Fa LI Ming-Yue WANG Juan YE Bo-Ping*
(School of Life Science and Technology, China Pharmaceutical University, Nanjing, Jiangsu 210009, China)
Abstract: In the present study, an endophytic fungus isolate FTJZZJ09, which isolated from the fresh bulbs of Fritillaria thunbergii Miq., was identified as Penicillium chrysogenum based on its morpho-logical characters and internal transcribed spacer (ITS) sequence. After being cultured in the modified CzapeK-DoX medium (3 g/L maltose, 3 g/L peptone A, 0.1 g/L K2HPO4, 0.05 g/L KCl, 0.3 g/L NaNO3, 0.05 g/L MgSO4·7H2O, 0.001 g/L FeSO4·7H2O, pH 6.5), it can secrete antibacterial metabolites under the condition of 28°C in a rotary shaker at 160 r/min for 7 days. Three antibacterial compounds were isolated from the ethyl acetate extract of the fermentation broth by silica gel, they were elucidated as cyclo (Pro-Gly), cyclo (Pro-Val) and 2-acetyl-4 (3H) quinazolinone. All the three compounds could inhibit the growth of Bacillus subtilis with the minimal inhibitory concentration (MIC) value of 0.8, 0.8, and 0.4 g/L respectively, while they showed no apparent effects against the growth of Gram-negative bacteria.
Keywords: Fritillaria thunbergii, Endophytic fungus, Penicillium chrysogenum, Diketopiperazines, Quinazolinones
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