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Production of the biotechnologically relevant AFP from Aspergillus giganteus in the yeast Pichia pastoris
- Source :
- E-Prints Complutense. Archivo Institucional de la UCM, instname, E-Prints Complutense: Archivo Institucional de la UCM, Universidad Complutense de Madrid, Digital.CSIC. Repositorio Institucional del CSIC
- Publication Year :
- 2010
- Publisher :
- Elsevier, 2010.
-
Abstract
- El pdf del artículo es la versión pre-print.<br />The mould Aspergillus giganteus produces a basic, low molecular weight protein (AFP) showing in vitro and in vivo antifungal properties against important plant pathogens. AFP is secreted as an inactive precursor containing an amino-terminal extension of six amino acids (If-AFP) which is later removed to produce the active protein. The molecular basis to explain this behavior and the features that determine the fungal specificity of this protein are not completely solved. In this work, the mature AFP (AFP*) and a version of AFP with an extended amino-terminal (proAFP) have been cloned and produced in the yeast Pichia pastoris. The two proteins have been purified to homogeneity and characterized from structural and functional points of view. AFP* produced is practically indistinguishable from the natural fungal protein in terms of its spectroscopic and antifungal properties while proAFP is mostly inactive under identical assay conditions. The availability of an active AFP protein produced in P. pastoris will allow getting further insights into the mode of action and targeting specificity of AFP by using site-directed mutagenesis approaches.<br />This work was supported by Grants BFU2006- 04404 and BIO2006-05583 from the Ministerio de Educación y Ciencia (MEC) (Spain) and within the center CONSOLIDER on Agrigenomics (MEC) and the Xarxa de referencia en Biotecnología (Generalitat de Catalunya).
- Subjects :
- Bioquímica
Antifungal Agents
Molecular Sequence Data
Mutagenesis (molecular biology technique)
Peptide
Pichia
Pichia pastoris
Fungal Proteins
Fusarium
Amino Acid Sequence
Cysteine-rich
Peptide sequence
neoplasms
chemistry.chemical_classification
Fungal protein
Antifungals
Biología molecular
biology
Base Sequence
digestive, oral, and skin physiology
biology.organism_classification
Yeast
digestive system diseases
Amino acid
Aspergillus
chemistry
Biochemistry
embryonic structures
Antimicrobial
Biotechnology
Subjects
Details
- Language :
- Spanish; Castilian
- Database :
- OpenAIRE
- Journal :
- E-Prints Complutense. Archivo Institucional de la UCM, instname, E-Prints Complutense: Archivo Institucional de la UCM, Universidad Complutense de Madrid, Digital.CSIC. Repositorio Institucional del CSIC
- Accession number :
- edsair.doi.dedup.....817a886e4ffdab72774bae250a01aea3