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4. Discussion The global antifungal market was estimated at $9.4 billion in 2010 and is expected to grow at a rate of 1.9% during 2010-2017. The major class of antifungal compound includes azoles, polyenes, pyrimidines, allyamines and echinodians. The antifungal therapies intend eradicating fungi through several action mechanisms, mainly involving destruction of cell wall and inhibition of cell division. There is a significant unmet need of novel anti- fungal compounds, because the present portfolio of treatments interact unfavorably with other medication, have resistance problem, low spectrum of activity, are fungistatic as opposed to fungicidal and are often toxic. Hence research has been focused on development of potent and safe antifungal compounds, either synthetic or from microbial sources [24]. In the present study, we exploited the marine resources for isolation of actinomycetes strains producing antifungal compounds. In the course of study, the marine invertebrate associated active acti- nomycetes strain PM0525875 was isolated and identified as Actinoalloteichus cyanogriseus. The antifungal principle from this strain was characterized as Caerulomycin A that showed potent in-vitro activity against pathogenic and drug resistant Candida strains. Caerulomycin A was first isolated from S. caeruleus [25] [26] as an antibiotic. Later it was reported to have anti-cancer activity [27]. In-vitro and in-vivo activity of Caerulomycin A against Entamoeba histolytica has been reported [28]. Natural derivatives of Caerulomycin A has also been reported [29]. Caerulomycin B and C, a new 2,2′-dipyridyl derivatives were isolated from Streptomyces caeruleus supplemented with 1 mM L-tryptophan. This group also reported the isolation of Caerulomycin D [19]. Additional natural derivatives of Caerulomycin F-K were isolated from marine derived actinomycetes (Actinoalloteichus cyanogriseus) that showed anticancer and antibacterial activity [29]. Lin et al. identified the biosynthetic gene cluster for Caerulomycin A from ma- rine actinomycetes Actinoalloteichus cyanogriseus. It mainly consists of unusual hybrid polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) [30]. The biosynthesis of Caerulomycin A based on enzy- matic modifications leading to structural changes, have been studied [31]. Chemical synthesis of Caerulomycin A-C has also been reported [32] [33]. The immunosuppressive activity of Caerulomycin A and its natural ana- logues has been reported [34]. Although Caerulomycin A has not been entered in clinic so far, may be due to its limitations, it could be the ideal molecule for generating semi synthetic druggable derivatives having potential application as an anticancer, antifungal or immunosuppressive drug. The strain and the optimized fermentation process described here could be useful for large scale production of this molecule for further drug development studies. 5. Conclusion In our in-house screening program in the quest of novel and superior antifungal compounds, an actinomycetes strain PM0525875 was isolated from a marine invertebrate. The active principle was characterized to be Caeru- lomycin A. Minimum inhibitory concentration (MIC) of the compound was found in the range of 0.39 - 1.56 μg/ml against pathogenic fungal test strains. The phylogenetic analysis of producer strain using 16S rRNA se- quence showed closest match with Actinoalloateichus cyanogriseus. Overall, Caerulomycin A was isolated from marine invertebrate-associated Actinoalloteichus sp. using optimized medium and fermentation conditions. |
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bxzc123: 金幣+60, 翻譯EPI+1, ★★★很有幫助 2014-10-22 12:43:39
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bxzc123: 金幣+60, 翻譯EPI+1, ★★★很有幫助 2014-10-22 12:43:39
bxzc123: 金幣+40, ★★★很有幫助 2014-10-22 20:39:11
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4 討論 2010年全球抗真菌市場估計值為94億美元，且在2010至2017年間有望以1.9%的增長率持續(xù)增長。 抗菌劑的主要成分含唑類、多烯類、嘧啶類、以及烯胺類 �？拐婢煼ㄔ噲D通過多種作用機制—主要是破壞細胞壁及抑制細胞的分解—來根除真菌。新型抗真菌化合物的需求量遠未得到滿足，其原因是現(xiàn)有的組合療法內部機制間存在相互抵制，且作用范圍小，其與抗菌劑的作用相悖，甚至會使抗菌劑失效。 因而有研究致力于開發(fā)高效、安全的抗真菌化合物，包括合成及從細菌中提取[24]。在本研究中，我們開發(fā)利用了細菌源進行放線菌株的隔離，來制取抗真菌化合物。我們在試驗中將海洋無脊椎動物的相關抗菌性菌株進行了隔離，并確定其為海洋來源放線菌。該菌株的抗菌性原理類似于淺藍霉素 A，其對致病性和耐藥性念珠菌菌株表現(xiàn)出很強的體外活性。 淺藍霉素 A是一種首次由S. caeruleus [25] [26]分離的抗生素。之后有報道稱其有抗癌作用[27]。淺藍霉素 A的體內抗痢疾變形蟲作用已經報道[28]。還有報道指出淺藍霉素 A的人工合成方法[29]。 淺藍霉素B與C是由青藍鏈霉菌補充以1mM左旋色氨酸分離而得的一種新型聯(lián)吡啶衍生物。該課題組還報道了淺藍霉素D 的分離提取[19]。其他自然衍生物淺藍霉素F-K均分離自海洋來源放線菌(稱為Actinoalloteichus cyanogriseus)，具有抗癌性及抗菌性[29]。 Lin等鑒定了分離自海洋放線菌Actinoalloteichus cyanogriseus的淺藍霉素A的生物基因組。結果發(fā)現(xiàn)該淺藍霉素A主要含不常見的雜聚酮合酶（PKS）及無核糖體型肽合成酶（NRPS）[30]. 淺藍霉素A 的生物合成以對酶的修飾為基礎實現(xiàn)其結構的改變，已有相關研究[31].淺藍霉素A-C的化學合成法已有報道[32][33]. 淺藍霉素A免疫抑制性及其自然界類似物已有報道[34]。雖然淺藍霉素A至今還未用于臨床——可能由于其限制性——但其仍可作為制取半合成藥物衍生物的理想分子，將成為一種有應用潛力的抗癌、殺菌或免疫抑制性的藥物。 5 結論 我們?yōu)榱双@得新型、優(yōu)化的抗真菌化合物設定了室內篩選程序，利用該程序從一種海洋無脊椎動物中分離出一段放線菌主鏈PM0525875。其活性成分為淺藍霉素A。該化合物經對抗致病真菌的測試，得出其最低抑菌濃度為0.39-1.56μg/ml。利用16S rRNA對產生菌進行系統(tǒng)分析，結果表明其與Actinoalloteichus cyanogriseus（海洋來源放線菌）有相關匹配性。綜上所述，本研究利用最優(yōu)方法及最佳發(fā)酵條件，從海洋無脊椎聯(lián)合放線菌淺藍霉素A中分離得到了淺藍霉素A。 |
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