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Chitinase Production by Trichoderma harzianum Grown on a Chitin-Rich Mushroom Byproduct Formulated Medium

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Abstract

Purpose

The aim of this work was to evaluate the use of chitin-rich mushroom (Agaricus bisporus) by-products chitin/glucan enriched fraction (M-Ch/G-F) as main carbon source for the production of chitinases by three different microorganisms (Trichoderma harzianum, Trichoderma atroviride and Bacillus licheniformis), in an attempt to obtain these enzymes using a cheap and abundant fermentation medium.

Methods

Microorganisms were grown in submerged fermentation using different media formulated with chitin powder (Chp), colloidal chitin (Chc) and M-Ch/G-F as the main carbon source, respectively. Enzyme productivity and secretion (secretome) was studied by electrophoretic and proteomic methods.

Results

All microorganisms produced higher chitinase activity in a medium formulated with M-Ch/G-F as a carbon source compared to medium formulated with Chp or CHc. T. harzianum showed the highest chitinase productivity (261.5 mU L−1 per day). Chitinase production was monitored by electrophoretic and proteomic methods. Electrophoretic method allowed the detection of 28 different proteins—three different chitinases with 82, 50 and 31 kDa. Proteomic analysis could identify 161 different proteins: 60 of them hydrolases, and 80% having glycolytic activity—5 of them were chitinases. These results show that cultivation of T. harzianum in a cheap and abundant fermentation medium represents a good procedure for large scale production of chitinases.

Conclusions

Our results show that cultivation of T. harzianum in a culture medium formulated with M-Ch/G-F, a cheap and abundant fermentation medium, is a good procedure for large scale production of glycosidases, particularly chitinases within a relatively short cultivation period of 6 days.

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Abbreviations

M-Ch/G-F:

Mushroom chitin/glucan enriched fraction

Mm:

Minimal medium

GlcNAc:

N-acetylglucosamine

Chp :

Chitin powder

Chc :

Colloidal chitin

PDB:

Potato dextrose broth

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Acknowledgements

We are grateful to the Spanish Ministry of Science and Innovation for the financial support of this work (Project RTC-2015-4039-2), which has partial financial support from the FEDER funds of the European Union. J.A. del Campo was supported by Nicolás Monardes Program from Servicio Andaluz de Salud (SAS). The authors thank Paula Bautista for assistance in the preparation of the manuscript.

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Authors and Affiliations

  1. Departamento de Bioquímica y Biología Molecular, Universidad de Sevilla, C/Profesor García González 2, 41012, Sevilla, Spain

    Anabell del Rocío Urbina-Salazar, Alberto Renato Inca-Torres, Gonzalo Falcón-García, Pilar Carbonero-Aguilar, Bruno Rodríguez-Morgado, Juan Parrado & Juan Bautista

  2. Facultad de Ciencias, Escuela Superior Politécnica de Chimborazo, Panamericana Sur km 1 1/2, Riobamba, Ecuador

    Anabell del Rocío Urbina-Salazar & Alberto Renato Inca-Torres

  3. CIBERehd, Hospital Universitario de Valme, 41014, Sevilla, Spain

    José A. del Campo

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  1. Anabell del Rocío Urbina-Salazar
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Correspondence to Juan Bautista.

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Urbina-Salazar, A., Inca-Torres, A.R., Falcón-García, G. et al. Chitinase Production by Trichoderma harzianum Grown on a Chitin-Rich Mushroom Byproduct Formulated Medium. Waste Biomass Valor 10, 2915–2923 (2019). https://doi.org/10.1007/s12649-018-0328-4

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