Production of recombinant bleaching enzymes from thermophilic microorganisms in fungal hosts.
Cost-effective production of enzymes for industrial processes makes the appropriate selection of the host-vector expression system critical. We have developed two systems for the bulk production of bleaching enzymes from thermophiles. Kluyveromyces lactis has been developed as a secretion host employing expression vectors based on the 2mu-like plasmid pKD1 of Kluyveromyces drosophilarium. Our second system involves the filamentous fungus Trichoderma reesei. Fusion and nonfusion vectors have been constructed using the strong cellobiohydrolase 1 (cbh1) promoter. The KEX2 protease cleavage site and a 6 x HIS-tag have been incorporated to facilitate both cleavage and purification of the mature foreign proteins.
Bergquist
Peter L
PL
Department of Biological Sciences, Macquarie University, Sydney, NSW, Australia. peter.bergquist@mq.edu.au
Te'o
V S Junior
VS
Gibbs
Moreland D
MD
Cziferszky
Angela C E
AC
De Faria
Fabricia P
FP
Azevedo
Maristela O
MO
Nevalainen
K M Helena
KM
D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology; D001709:Biotechnology / Q000379:methods; D004798:Enzymes / Q000235:genetics* / Q000302:isolation & purification; D005658:Fungi / Q000201:enzymology* / Q000235:genetics; D006358:Hot Temperature; D007716:Kluyveromyces / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011487:Protein Conformation; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D014242:Trichoderma / Q000201:enzymology; D043325:Xylan Endo-1,3-beta-Xylosidase; D014995:Xylosidases / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D004798:Enzymes; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D014995:Xylosidases; D043325:Xylan Endo-1,3-beta-Xylosidase