Selective targeting of a laccase from Stachybotrys chartarum covalently linked to a carotenoid-binding peptide.
Atwo-step targeting strategy was used to identify improved laccases for bleaching carotenoid-containing stains on fabric. We first applied a modified phage display technique to identify peptide sequences capable of binding specifically to carotenoid stains and not to fabric. Prior deselection on the support on which the carotenoid was localized, increased stringency during the biopanning target selection process, and analysis of the phage peptides' binding to the target after acid elution and polymerase chain reaction (PCR) postacid elution, were used to isolate phage peptide libraries with increased binding selectivity and affinity. Peptide sequences were selected based on identified consensus motifs. We verified the enhanced carotenoid-binding properties of the peptide YGYLPSR and subsequently cloned and expressed C-terminal variants of laccase from Stachybotrys chartarum containing carotenoid-binding peptides YGYLPSR, IERSAPATAPPP, KASAPAL, CKASAPALC, and SLLNATK. These targeted peptide-laccase fusions demonstrate enhanced catalytic properties on stained fabrics.
Janssen
G G
GG
Genencor International, Inc., 925 Page Mill Road, Palo Alto, CA 94304, USA. ggjanssen@netzero.com
Baldwin
T M
TM
Winetzky
D S
DS
Tierney
L M
LM
Wang
H
H
Murray
C J
CJ
D002338:Carotenoids / Q000737:chemistry*; D004396:Coloring Agents / Q000737:chemistry; D042845:Laccase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011485:Protein Binding / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013191:Stachybotrys / Q000201:enzymology* / Q000235:genetics
D004396:Coloring Agents; D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins; D002338:Carotenoids; D042845:Laccase