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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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讨论 2010年的评估指出抗菌剂的全球市场值为94亿美元，而且从2010年至2017年间会以1.9%的速率不断增长。抗菌剂的主要成分含唑类、多烯类、嘧啶类、以及烯胺类 。抗真菌疗法试图通过多种作用机制—主要是破坏细胞壁及抑制细胞的分解—来根除真菌。新型抗真菌化合物的需求量远未得到满足，其原因是现有的组合疗法内部机制间存在相互抵制，且作用范围小，其与抗菌剂的作用相悖，甚至会使抗菌剂失效。 因而有研究致力于开发高效、安全的抗真菌化合物，包括合成及从细菌中提取[24]。在本研究中，我们开发利用了细菌源进行放线菌株的隔离，来制取抗真菌化合物。我们在试验中将海洋无脊椎动物的相关抗菌性菌株进行了隔离，并确定其为海洋来源放线菌。该菌株的抗菌性原理类似于浅蓝霉素 A，其对致病性和耐药性念珠菌菌株表现出很强的体外活性。 浅蓝霉素 A是一种首次由S. caeruleus [25] [26]分离的抗生素。之后有报道称其有抗癌作用[27]。浅蓝霉素 A的体内抗痢疾变形虫作用已经报道[28]。还有报道指出浅蓝霉素 A的人工合成方法[29]。  |
2楼2014-10-17 20:42:24