Vol.:(0123456789) 1 3
Waste and Biomass Valorization
https://doi.org/10.1007/s12649-018-0328-4
ORIGINAL PAPER
Chitinase Production by Trichoderma harzianum Grown on a Chitin-
Rich Mushroom Byproduct Formulated Medium
Anabell del Rocío Urbina‑Salazar
1,2
· Alberto Renato Inca‑Torres
1,2
· Gonzalo Falcón‑García
1
·
Pilar Carbonero‑Aguilar
1
· Bruno Rodríguez‑Morgado
1
· José A. del Campo
3
· Juan Parrado
1
·
Juan Bautista
1
Received: 24 January 2018 / Accepted: 7 May 2018
© Springer Science+Business Media B.V., part of Springer Nature 2018
Abstract
Purpose The aim of this work was to evaluate the use of chitin-rich mushroom (Agaricus bisporus) by-products chitin/glu-
can enriched fraction (M-Ch/G-F) as main carbon source for the production of chitinases by three diferent 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 diferent media formulated with chitin powder (Ch
p
),
colloidal chitin (Ch
c
) 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 Ch
p
or CH
c
. 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 diferent proteins—three diferent chitinases with 82, 50 and 31 kDa. Proteomic analysis could identify
161 diferent 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.
Graphical Abstract
Keywords Chitin-rich mushroom byproduct · Trichoderma harzianum · Bacillus licheniformis · Agaricus bisporus ·
Chitinase
Abbreviations
M-Ch/G-F Mushroom chitin/glucan enriched fraction
Mm Minimal medium
GlcNAc N-acetylglucosamine
Ch
p
Chitin powder
* Juan Bautista
jdbaut@us.es
Extended author information available on the last page of the article