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Improved pectinase production in Penicillium griseoroseum recombinant strains

Author:
Teixeira, J.A., Gonçalves, D.B., de Queiroz, M.V., de Araújo, E.F.
Source:
Journal of applied microbiology 2011 v.111 no.4 pp. 818-825
ISSN:
1364-5072
Subject:
Aspergillus nidulans, Penicillium, Southern blotting, carbon, fungi, genes, industry, juices, pectin lyase, pectins, plasmids, polygalacturonase, protein synthesis, proteolysis, secretion, wines
Abstract:
Aims: To obtain recombinant strains of Penicillium griseoroseum that produce high levels of pectin lyase (PL) and polygalacturonase (PG) simultaneously. Methods and Results: A strain with high production of PL was transformed with the plasmid pAN52pgg2, containing the gene encoding PG of P. griseoroseum, under control of the gpd promoter gene from Aspergillus nidulans. Southern blot analysis demonstrated that all strain had at least one copy of pAN52pgg2 integrated into the genome. The recombinant strain P. griseoroseum T20 produced levels of PL and PG that were 266‐ and 27‐fold greater, respectively, than the wild‐type strain. Furthermore, the extracellular protein profile of recombinant T20 showed two protein bands of c. 36 and 38 kDa, associated with PL and PG, respectively. Conclusions: This recombinant strain T20 produces PL and PG using carbon sources of low costs, and an enzyme preparation that is free of cellulolytic and proteolytic activities. Significance and Impact of the Study: PL and PG play an important role in the degradation of pectin. Owing to their use in the juice and wines industries, there is a growing interest in the inexpensive production of these enzymes. This work describes an efficient system of protein expression and secretion using the fungus P. griseoroseum.
Agid:
105197