title: Heterologous production of a laccase from the basidiomycete Pycnoporus cinnabarinus in the dimorphic yeast Yarrowia lipolytica.
abstract: Pycnoporus cinnabarinus lac1 gene was expressed in Yarrowia lipolytica. Different secretion signals and culture media were tested. Production was correlated to both culture growth rate and cell morphology (highest at low growth rate, without mycelium). Recombinant laccase was characterized (immunodetection, N-terminal sequencing) and purified. Production was estimated to 20 mgl(-1) in a bioreactor. Thus, complex metalloenzymes can be produced in Yarrowia, assuming some control of host physiology. Lac1p production was compared in Yarrowia, Pichia and Aspergillus: recombinant proteins were active, but host systems differed in transformation efficiency, production, and glycosylation. If not the best producer, Yarrowia offers very high transformation efficiencies, allowing the genetic engineering of laccases for industrial applications.
authors.affiliation: UMR Microbiologie et Génétique Moléculaire INRA/CNRS/INAP-G, Centre de Biotechnologie Agro-Industrielle, BP 01, 78850 Thiverval-Grignon, France. madzak@grignon.inra.fr
authors.forename: Catherine
authors.initials: C
authors.lastname: Madzak
authors.forename: Ludovic
authors.initials: L
authors.lastname: Otterbein
authors.forename: Mohamed
authors.initials: M
authors.lastname: Chamkha
authors.forename: Serge
authors.initials: S
authors.lastname: Moukha
authors.forename: Marcel
authors.initials: M
authors.lastname: Asther
authors.forename: Claude
authors.initials: C
authors.lastname: Gaillardin
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Beckerich
mesh_terms: D001487:Basidiomycota / Q000201:enzymology* / Q000235:genetics; D003470:Culture Media; D007218:Industrial Microbiology; D042845:Laccase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics*
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D042845:Laccase