Dos Santos Juliana A, Vieira Juliana M F, Videira Alexandre, Meirelles Lucas A, Rodrigues André, Taniwaki Marta H, Sette Lara D
Departamento de Bioquímica e Microbiologia, Universidade Estadual Paulista Júlio de Mesquita Filho-UNESP, 24A, 1515, Rio Claro, SP, 13506-900, Brazil.
Instituto de Tecnologia de Alimentos-ITAL, Campinas, SP, Brazil.
AMB Express. 2016 Mar;6(1):25. doi: 10.1186/s13568-016-0194-z. Epub 2016 Mar 24.
Marine-derived fungi have been reported as relevant producers of enzymes, which can have different properties in comparison with their terrestrial counterparts. The aim of the present study was to select from a collection of 493 marine-derived fungi the best producer of xylanase in order to evaluate the enzymatic production under different conditions. A total of 112 isolates produced xylanase in solid medium containing xylan as the carbon source, with 31 of them able to produce at least 10 U/mL of the enzyme. The best production (49.41 U/mL) was achieved by the strain LAMAI 31, identified as Aspergillus cf. tubingensis. After confirming the lack of pathogenicity (absence of ochratoxin A and fumonisin B2 production) this fungus was submitted to the experimental design in order to evaluate the effect of different variables on the enzymatic production, with the aim of optimizing culture conditions. Three experimental designs (two Plackett-Burman and one factorial fractional) were applied. The best condition for the enzymatic production was defined, resulting in an increase of 12.7 times in comparison with the initial production during the screening experiments. In the validation assay, the peak of xylanase production (561.59 U/mL) was obtained after 96 h of incubation, being the best specific activity achieved after 72 h of incubation. Xylanase from A. cf. tubingensis LAMAI 31 had optimum pH and temperature at 5.0 and 55 °C, respectively, and was shown to be stable at a range of 40-50 °C, and in pH from 3.6 to 7.0. Results from the present work indicate that A. cf. tubingensis LAMAI 31 can be considered as a new genetic resource for xylanase production.
据报道,海洋来源的真菌是酶的重要生产者,与陆地同类真菌相比,它们可能具有不同的特性。本研究的目的是从493株海洋来源的真菌中筛选出木聚糖酶的最佳生产者,以便在不同条件下评估酶的产量。共有112株分离物在以木聚糖为碳源的固体培养基中产生木聚糖酶,其中31株能够产生至少10 U/mL的该酶。菌株LAMAI 31产量最高(49.41 U/mL),鉴定为烟曲霉复合种(Aspergillus cf. tubingensis)。在确认其无致病性(不产生赭曲霉毒素A和伏马菌素B2)后,将该真菌进行实验设计,以评估不同变量对酶产量的影响,目的是优化培养条件。应用了三个实验设计(两个Plackett-Burman设计和一个析因分数设计)。确定了酶生产的最佳条件,与筛选实验中的初始产量相比,产量提高了12.7倍。在验证试验中,培养96小时后获得木聚糖酶产量峰值(561.59 U/mL),培养72小时后获得最佳比活性。烟曲霉复合种LAMAI 31的木聚糖酶的最适pH和温度分别为5.0和55℃,并且在40-50℃范围内以及pH值为3.6至7.0时表现出稳定性。本研究结果表明,烟曲霉复合种LAMAI 31可被视为木聚糖酶生产的新遗传资源。