Instituto de Ciências Agrárias (ICIAG), Universidade Federal de Uberlândia, Uberlândia, Brazil.
N Biotechnol. 2012 Feb 15;29(3):451-6. doi: 10.1016/j.nbt.2011.05.004. Epub 2011 Jun 12.
This study investigated the fungi diversity of fresh olive (Olea europaea L.) fruits, olive paste (crushed olives) and olive pomace (solid waste) and screened and quantified enzymatic activities with biotechnological applications. Fungi were randomly isolated from olive cultivars from Castilla La Mancha region (Spain). Identification included comparison of their polymerase chain reaction (PCR) amplicons of the ITS1-5.8S-ITS2 ribosomal DNA region, followed by nucleotide sequence analysis. Fourteen different species with DNA sequences of different similarities were identified, belonging to seven different genera (Aspergillus, Penicillium, Rhizomucor, Mucor, Rhizopus, Lichtheimia and Galactomyces). Aspergillus fumigatus, followed by Galactomyces geotrichum, Penicillium commune and Rhizomucor variabilis var. regularior were the most frequent species. Specific enzyme screening was assayed on agar plates, using cellobiose, carboxymethylcellulose (CMC), polygalacturonic acid and CaCl(2)/Tween 80 as substrates for β-glucosidase, carboxymethylcellulase (CMCase), polygalacturonase and lipase, respectively. Species exhibiting the best activities were: Aspergillus fumigatus (for β-glucosidase, CMCase and lipase); Rhizopus oryzae (for β-glucosidase and lipase); Rhizomucor variabilis (for β-glucosidase, CMCase and polygalacturonase); Mucor fragilis (β-glucosidase, CMCase and lipase); Galactomyces geotrichum (for β-glucosidase, polygalacturonase and lipase) and Penicillium commune and Penicillium crustosum (for lipase). The species that had shown the best enzymatic activities were grown on hemicellulose, cellulose and pectin and some activities were quantified (xylanase, cellulase, β-glucosidase and pectinase). An isolate of A. fumigatus and one of A. niger showed the best cellulase and xylanase activities, while no species presented good pectinase and β-glucosidase activities. The selected species with potential enzymatic activities could be used for future applications of industrial interest.
本研究调查了新鲜橄榄（Olea europaea L.）果实、橄榄糊（压碎的橄榄）和橄榄渣（固体废料）中的真菌多样性，并筛选和量化了具有生物技术应用的酶活性。真菌是从卡斯蒂利亚拉曼恰地区（西班牙）的橄榄品种中随机分离出来的。鉴定包括比较其 ITS1-5.8S-ITS2 核糖体 DNA 区域的聚合酶链反应（PCR）扩增子，然后进行核苷酸序列分析。鉴定出了 14 种不同相似度的不同种，属于 7 个不同属（曲霉属、青霉属、根毛霉属、毛霉属、根霉属、亮白菌属和毕赤酵母属）。曲霉属烟曲霉、随后是毕赤酵母属酒香酵母、青霉属共同青霉和根毛霉属可变根毛霉是最常见的物种。在琼脂平板上，用纤维二糖、羧甲基纤维素（CMC）、聚半乳糖醛酸和 CaCl2/Tween 80 作为β-葡萄糖苷酶、羧甲基纤维素酶（CMCase）、聚半乳糖醛酸酶和脂肪酶的底物，对特定的酶进行了筛选。表现出最佳活性的物种为：烟曲霉（β-葡萄糖苷酶、CMCase 和脂肪酶）；米根霉（β-葡萄糖苷酶和脂肪酶）；可变根毛霉（β-葡萄糖苷酶、CMCase 和聚半乳糖醛酸酶）；脆弱毛霉（β-葡萄糖苷酶、CMCase 和脂肪酶）；酒香酵母（β-葡萄糖苷酶、聚半乳糖醛酸酶和脂肪酶）和青霉属共同青霉和青霉属皮落青霉（脂肪酶）。表现出最佳酶活性的物种在半纤维素、纤维素和果胶上生长，并对一些活性进行了量化（木聚糖酶、纤维素酶、β-葡萄糖苷酶和果胶酶）。烟曲霉和黑曲霉的一个分离株表现出最好的纤维素酶和木聚糖酶活性，而没有一种具有良好的果胶酶和β-葡萄糖苷酶活性。具有潜在酶活性的选定物种可用于未来具有工业应用前景的应用。
