Estudio de la producción de quitinasas, proteasas e hidrofobinas de Lecanicillium lecanii en cultivos líquido y sólido utilizando diversas fuentes de carbono
Entomopathogenic fungi produce various enzymes during the process of pathogenesis, including proteases and chitinases, as well as allowing hydrophobins its adherence to the host, which has been reported that extrinsic factors affecting its production (St. Leger et al, 1986 y 1998 ). However, there a...
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| Tipo de recurso: | tesis de maestría |
| Estado: | Versión publicada |
| Fecha de publicación: | 2009 |
| País: | México |
| Institución: | Universidad Autónoma Metropolitana |
| Repositorio: | Repositorio Institucional de la UAM Iztapalapa |
| Idioma: | español |
| OAI Identifier: | oai:bindani.izt.uam.mx:t148fh43j |
| Acceso en línea: | https://doi.org/10.24275/uami.t148fh43j |
| Access Level: | acceso abierto |
| Palabra clave: | info:eu-repo/classification/LEM/Fungi Pathogenesis info:eu-repo/classification/LEM/Hongos -- Patogénesis info:eu-repo/classification/LEM/Enzymes info:eu-repo/classification/LEM/Enzimas info:eu-repo/classification/LEM/Chitinase info:eu-repo/classification/LEM/Quitinasas info:eu-repo/classification/cti/2 |
| Sumario: | Entomopathogenic fungi produce various enzymes during the process of pathogenesis, including proteases and chitinases, as well as allowing hydrophobins its adherence to the host, which has been reported that extrinsic factors affecting its production (St. Leger et al, 1986 y 1998 ). However, there are not studies about their production by fungi such as Lecanicillium lecanii, an entomopathogenic fungus which is used as bio-insecticide in agriculture and horticulture on a commercial level. The aim of this study was the evaluation of the effect of culture type; on solid media (SSC) using polyurethane as support, and on media submerged (SmC), and the carbon sources, in the production of chitinolytic enzymes and hydrophobins of Lecanicillium lecanii. This fungus was grown on SSC media and SmC, as carbon sources were used: fructose (F), fructose with chitin (CF), sucrose with chitin (SaQ) and chitin (Q). Increased production of Nacetylhexosaminidase (Nhasa) was obtained using colloidal chitin at 72 h in SmC and SSC cultures. The Nhasa production in SSCQ (17.7 U/g initial substrate) was higher than the culture SmCQ (1.3 U/g initial substrate). The basic (6.9 U/µg protein) and acid (6.3 U/µg protein) proteases production was similar in presence of chitin, while in cultures with others carbon sources, the acid protease production was 10-fold higher than basic proteases. The protease production (acid: 1.6 U/µg protein y basic: 1 U/µg protein) was 6-fold lower in solid culture than in SmC, which affects the production of other proteins since might be hydrolyzed by the proteases. Hydrophobins production was 10-fold higher in SSC cultivation than in SmC. In addition, the extract of hydrophobins of SSCQ showed higher activity on surface due to the reduction up to 50% of the hydrophobicity of teflon, whereas hydrophobins found in SmCF reduces only 8% without significant difference with teflon. The molecular mass (Mr) of these proteins were determined ca. 7 kDa. The zymograms showed bands with chitinolytic activity with molecular mass, from 9 to 111 kDa. We determined the presence of endochitinases, exochitinases and N-acetylhexosaminidase in most treatments. Only SmC culture with added fructose and colloidal chitin showed bands chitobiase activity. These results provide information of the effects of culture type and carbon sources on production of proteases, chitinases and hydrophobins of L. lecanii, which are products of great importance due to their potential applications. It is important to note that hydrophobins were produced at high levels under favorable conditions for the induction of chitinases such as colloidal chitin and solid culture that suggest a relationship that might be related to the fungi mechanism of pathogenesis, thus more research should be performed. |
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