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在分批式生物反应器中,子囊菌共培养物在纤维素酶生产方面优于混合培养物。

An ascomycota coculture in batch bioreactor is better than polycultures for cellulase production.

作者信息

Hernández Christian, Milagres Adriane M F, Vázquez-Marrufo Gerardo, Muñoz-Páez Karla María, García-Pérez José Antonio, Alarcón Enrique

机构信息

Instituto de Biotecnología y Ecología Aplicada (INBIOTECA), Universidad Veracruzana, Avenida de las culturas veracruzanas no. 101, colonia Emiliano Zapata, 91090, Xalapa, Veracruz, Mexico.

Departamento de Biotecnología, Escola de engenharia de Lorena (EEL), Universidade de São Paulo, Estrada Municipal do Campinho s/n - Pte Nova, Lorena, SP, 12602-810, Brazil.

出版信息

Folia Microbiol (Praha). 2018 Jul;63(4):467-478. doi: 10.1007/s12223-018-0588-1. Epub 2018 Feb 8.

Abstract

Efficient hydrolysis of holocellulose depends on a proper balance between cellulase (endoglucanase, exoglucanase, β-glucosidase) and xylanase activities. The present study aimed to induce the production of cellulases and xylanases using liquid cultures (one, two, three, and four fungal strains on the same bioreactor) of wild strains of Trichoderma harzianum, Aspergillus niger, and Fusarium oxysporum. The strains were identified by amplification and analysis of the ITS rDNA region and the obtained sequences were deposited in Genbank. Enzymes (endoglucanase, exoglucansae, β-glucosidase, and xylanase activities) and the profile of extracellular protein isoforms (SDS-PAGE) produced by different fungal combinations (N = 14) were analyzed by Pearson's correlation matrix and principal component analysis (PCA). According to our results, induction of endoglucanase (19.02%) and β-glucosidase (6.35%) were obtained after 4 days when A. niger and F. oxysporum were cocultured. The combination of A. niger-T. harzianum produced higher endoglucanase in a shorter time than monocultures. On the contrary, when more than two strains were cultured in the same reactor, the relationships of competition were established, trending to diminish the amount of enzymes and the extracellular protein isoforms produced. The xylanase production was sensible to stress produced by mixed cultures, decreasing their activity. This is important when the aim is to produce cellulase-free xylanase. In addition, exoglucanase activity did not change in the combinations tested.

摘要

全纤维素的高效水解取决于纤维素酶(内切葡聚糖酶、外切葡聚糖酶、β-葡萄糖苷酶)和木聚糖酶活性之间的适当平衡。本研究旨在利用哈茨木霉、黑曲霉和尖孢镰刀菌野生菌株的液体培养物(在同一生物反应器中培养一、二、三、四种真菌菌株)诱导纤维素酶和木聚糖酶的产生。通过扩增和分析ITS rDNA区域对菌株进行鉴定,并将获得的序列保存在Genbank中。通过Pearson相关矩阵和主成分分析(PCA)分析了不同真菌组合(N = 14)产生的酶(内切葡聚糖酶、外切葡聚糖酶、β-葡萄糖苷酶和木聚糖酶活性)以及细胞外蛋白质同工型谱(SDS-PAGE)。根据我们的结果,黑曲霉和尖孢镰刀菌共培养4天后,内切葡聚糖酶(19.02%)和β-葡萄糖苷酶(6.35%)得到诱导。黑曲霉-哈茨木霉组合比单培养在更短时间内产生了更高的内切葡聚糖酶。相反,当在同一反应器中培养两种以上菌株时,会建立竞争关系,导致产生的酶量和细胞外蛋白质同工型减少。木聚糖酶的产生对混合培养产生的压力敏感,其活性降低。当目标是生产无纤维素酶的木聚糖酶时,这一点很重要。此外,在所测试的组合中外切葡聚糖酶活性没有变化。

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