Genomic gene encoding manganese peroxidase from a white-rot fungus Phanerochaete crassa WD1694
- PDF / 1,017,582 Bytes
- 8 Pages / 595.276 x 790.866 pts Page_size
- 34 Downloads / 178 Views
ORIGINAL ARTICLE
Genomic gene encoding manganese peroxidase from a white-rot fungus Phanerochaete crassa WD1694 Mariko Takano • Muneysohi Yamaguchi Hiroaki Sano • Masaya Nakamura • Hajime Shibuya • Yasumasa Miyazaki
•
Received: 19 June 2012 / Accepted: 26 October 2012 / Published online: 6 November 2012 Ó The Japan Wood Research Society 2012
Abstract The gene encoding manganese peroxidase of a white-rot fungus Phanerochaete crassa WD1694 was cloned and sequenced. Four genomic clones were sequenced in which 3 clones were existed as alleles. The analysis of intron–exon structures divided the 4 clones into three subfamilies that corresponded to mnp2 and mnp3 of Phanerochaete chrysosporium, and a new subfamily possessing only five introns. The purified P. crassa WD1694 MnP consisted of 4 isozymes with same molecular weight, same N-terminal sequence, and different pI. N-terminal sequence of deduced protein of P. crassa mnpB3 gene was identical to those of 4 MnP isozymes; however, the other 3 mnp genes had different N-terminal sequence. The molecular weight of encoded mature protein of mnpB3 gene and purified MnP had a gap that could be difference between MnP proteins encoded by single gene. The results suggested that 4 MnP isozymes of P. crassa WD1694 arose from single gene. Keywords Lignin Phanerochaete crassa Manganese peroxidase
Introduction White-rot fungus is the only organism that can effectively break down lignin, which is very resistant to general microbial attack. It is well known that extracellular peroxidases, such as LiP and MnP play major roles in the
lignin biodegradation process of white-rot fungi [1–4]. The catalytic mechanisms and molecular genetics of these ligninolytic peroxidases have been studied and have revealed their structural and functional properties [5–10]. It has also been reported that plant and fungal peroxidases may be arranged in a superfamily of three distinct classes; namely class I of bacterial and intracellular peroxidases, class II of fungal secretory peroxidases, and class III of secretory plant peroxidases, respectively [11]. The lignin-degrading peroxidases in class II of fungal secretory peroxidases have been further subdivided into three groups, LiP, MnP, and VP, based on the genetic and protein structural evidence [12]. Recently, phylogenetic analysis of fungal ligninolytic peroxidases was reported, showing the ubiquity and diversity of these enzymes among a wide range of ectomycorrhizal fungi, basidiomycetes and agaricomycetes [13–15]. We have studied the distribution of the extracellular peroxidase reaction of a white-rot fungus P. crassa WD1694 in detail, and showed how the MnP reaction occurred at the hyphal tips [16, 17]. Previously, we reported on the purification and characterization of MnP from P. crassa WD1694 [18]. In this report, we studied the cloning and sequence of genomic mnp genes of P. crassa WD1694 as an initial step toward understanding the molecular genetics of P. crassa WD1694.
Materials and methods Strains
M. Takano (&) M. Yamaguchi H. Sano
Data Loading...
