臺灣三種海洋細菌之生物活性成分研究

Studies on the bioactive constituents of three marine bacterial species isolated in Taiwan

許瑜真(Yu-chen Hsu)

指導教授 ： 李宗徽 博士

臺北醫學大學/藥學院/生藥學研究所/碩士(2009年)

摘要

以三種不同培養基篩選出143株海洋細菌株，經醱酵培養後，萃取液初步以抑制一氧化氮合成酶 (NOS) 的活性與利用Phenylephrine誘發大鼠 (Sprague-Dawley Rat) 胸主動脈收縮活性平台進行篩選出具有抑制血管收縮的菌株。結果自143株海洋菌株萃取液中發現二株對於RAW 264.7細胞具有細胞毒性，分別為 Streptomyces sp. (#HYC21) 和 Pseudoalteromonas sp. (#HYC13)，另一株 Pseudomonas aeruginosa (#M1B) 則具有抑制血管收縮的活性，後續針對這三株具有生物活性的菌株以PPY、PY和PYG 培養基分批擴大培養，進行一系列的分析、分離、純化與構造解析，計分離、決定出二十一個化合物，包括：二個環肽類抗生素 (cyclic peptide antibiotics) 的actinomycin X2 (1)、actinomycin D (2)；二個多溴酚類抗生素 (polybrominated phenolic antibiotics) 的 pentabromopseudilin (8) 和3,3′,5,5′-tetrabromo-2,2′-biphenyldiol (9)；三個膽酸及其類似物 (cholic acid and its analogs) 的cholic acid (10)、deoxycholic acid (11)、glycocholic acid (12)；五個喹啉生物鹼 (quinoline alkaloid) 分別為2- heptylquinol-4-one (13)、2-heptyl-3-hydroxyquinolin-4(1H)-one (14)、2-(1′E-nonenyl)quinol-4-one (15)、2-nonylquinol-4-one (16)、 3-heptyl-3-hydroxy-1,2,3,4-tetrahydroquinoline-2,4-dione (17)；二個酚類化合物 (phenolics) 的2-(2-hydroxyphenyl)-2-thiazoline-4-methanol (18)、4,4′-isopropylidenebisphenol (19)；一個吩嗪生物鹼 (phenazine alkaloid) phenazine-1-carboxamide (21) 及五個環化雙胺基酸 (diketopiperazine) 分別為cyclo-L-Pro-L-Val (3)、cyclo-L-Pro-L-Ile (4)、cyclo-L-Pro-L-Leu (5)、cyclo-D-Pro-L-Phe (6)、cyclo-L-Pro-L-Phe (7)、cyclo-L-Pro-L-Trp (20)。此外，由M1B所分離得到的化合物在生物活性及作用機制，刻正持續探究中。

關鍵字

海洋細菌

並列摘要

In this study, 143 strains of marine bacteria isolated from Taiwan were cultured for the screening of their inducible nitric oxide synthase (iNOS) inhibitory activity and vasorelaxing activity. Of these bacterial strains monitored, HYC21 (Streptomyces sp.) and HYC13 (Pseudoalteromonas sp.) exhibited cytotoxic activities against RAW 264.7 cell line. While M1B (Pseudomonas aeruginosa) exhibited vasorelaxing activity on Sprague-Dawley rats induced by phenylephrine. Based on these findings, the three strains were thus mass cultured and a series of separation and isolation were undertaken to investigate their active principles. Totally twenty-one compounds including 2 cyclic peptide antibiotics, 2 polybrominated phenolic antibiotics, 3 cholic acid and its analogs, 5 quinoline alkaloids, 2 phenolics, a phenazine alkaloid, and 6 diketopiperazine was isolated and identified. Their structures were elucidated to be actinomycin X2 (1), actinomycin D (2), cyclo-L-Pro-L-Val (3), cyclo-L-Pro-L-Ile (4), cyclo-L-Pro-L-Leu (5), cyclo-D-Pro-L-Phe (6), cyclo-L-Pro-L-Phe (7), pentabromopseudilin (8), 3,3′,5,5′-tetrabromo-2,2′-biphenyldiol (9), cholic acid (10), deoxycholic acid (11), glycocholic acid (12), 2-heptylquinol-4-one (13), 2-heptyl-3-hydroxyquinolin-4(1H)-one (14), 2- (1′E-nonenyl)quinol-4-one (15), 2-nonylquinol-4-one (16), 3- heptyl-3-hydroxy-1,2,3,4-tetrahydroquinoline-2,4-dione (17), 2-(2-hydroxyphenyl)-2-thiazoline-4-methanol (18), 4,4′-isopropylidenebisphenol (19), cyclo-L-Pro-L-Trp (20), phenazine-1-carboxamide (21).

並列關鍵字

Pseudomonas aeruginosa ； Actinomycin X2 ； diketopiperazine ； NOS

參考文獻

21. Blunt, J. W., Copp, B. R., Hu, W.-P., Munro, M. H. G., Northcote, P. T.,Prinsep, M. E. R., Marine natural products. Natural Product Reports 2009, 26, 170-244.

2. Roberts, J. M., Wheeler, A. J.,Freiwald, A., Reefs of the deep: the biology and geology of cold-water coral ecosystems. Science 2006, 312, 543-547.

4. Sibuet, M.,Olu, K., Biogeography, biodiversity and fluid dependence of deep-sea cold-seep communities at active and passive margins. Deep Sea Res. Part II: Topical Studies Oceanograph 1998, 45, 517-567.

6. Grassle, J. F.,Maciolek, N. J., Deep-sea species richness: regional and local diversity estimates from quantitative bottom samples. American Naturalist 1992, 139, 313-341.

7. Gage, J. D., Why are there so many species in deep-sea sediments? Journal of Experimental Marine Biology and Ecology 1996, 200, 257-286.