Manganese peroxidase from the white-rot fungus Phanerochaete chrysosporium is enzymatically active and accumulates to high levels in transgenic maize seed.
Manganese peroxidase (MnP) has been implicated in lignin degradation and thus has potential applications in pulp and paper bleaching, enzymatic remediation and the textile industry. Transgenic plants are an emerging protein expression platform that offer many advantages over traditional systems, in particular their potential for large-scale industrial enzyme production. Several plant expression vectors were created to evaluate the accumulation of MnP from the wood-rot fungus Phanerochaete chrysosporium in maize seed. We showed that cell wall targeting yielded full-length MnP, whereas cytoplasmic localization resulted in multiple truncated peroxidase polypeptides as detected by immunoblot analysis. In addition, the use of a seed-preferred promoter dramatically increased the expression levels and reduced the negative effects on plant health. Multiple independent transgenic lines were backcrossed with elite inbred corn lines for several generations with the maintenance of high-level expression, indicating genetic stability of the transgene.
Clough
Richard C
RC
ProdiGene, Inc., 101 Gateway Blvd., Suite 100, College Station, TX 77845, USA. clough@cox-internet.com
Pappu
Kameshwari
K
Thompson
Kevin
K
Beifuss
Katherine
K
Lane
Jeff
J
Delaney
Donna E
DE
Harkey
Robin
R
Drees
Carol
C
Howard
John A
JA
Hood
Elizabeth E
EE
D001673:Biodegradation, Environmental; D001709:Biotechnology; D002473:Cell Wall / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005800:Genes, Fungal; D008031:Lignin / Q000378:metabolism; D010544:Peroxidases / Q000235:genetics* / Q000378:metabolism*; D020075:Phanerochaete / Q000201:enzymology*; D012639:Seeds / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic; D003313:Zea mays / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D005656:Fungal Proteins; D008031:Lignin; D010544:Peroxidases; C051129:manganese peroxidase