Isoenzyme Multiplicity and Characterization of Recombinant Manganese Peroxidases from
            <i>Ceriporiopsis subvermispora</i>
            and
            <i>Phanerochaete chrysosporium</i>

Larrondo, Luis F.; Lobos, Sergio; Stewart, P. A. Hamilton; Cullen, Dan; Vicuña, Rafael

doi:10.1128/aem.67.5.2070-2075.2001

Cited by 33 publications

(24 citation statements)

References 32 publications

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“…The phenomenon of isoform multiplicity was previously observed with recombinant Cs-MnP1, as well as recombinant Phanerochaete chrysosporium MnP1, both expressed in A. nidulans (Larrondo et al, 2001). We have also observed multiple isoforms for P. chrysosporium multicopper oxidase when expressed in Aspergillus (L. F. Larrondo and others, unpublished).…”

Section: Discussionsupporting

confidence: 49%

“…Zymograms were as previously reported (Larrondo et al, 2001). Briefly, samples were applied with nonreducing denaturing loading buffer without boiling and subjected to SDS-PAGE at 4˚C.…”

Section: Methodsmentioning

confidence: 99%

“…nidulans A722 and A. niger A969 cotransformations were as described by Larrondo et al (2001), except that A. niger protoplasts were prepared from germinated conidia after an overnight incubation at 300 r.p.m. at 30˚C.…”

Section: Methodsmentioning

confidence: 99%

“…Analytical IEF and protein concentration measurements were as described by Larrondo et al (2001); RNA extraction and Northern blot hybridization were as described by Karahanian et al (1998). The DNA probe corresponding to the entire coding region of Lcs-1 was labelled with [a-32 P]dCTP by nick translation (Gibco-BRL).…”

Section: Methodsmentioning

confidence: 99%

“…K m values for ABTS were determined with 0?01 U laccase, using Eadie-Hofstee plots. For endoglycosidase treatment, 5 mg enzyme was treated as described by Larrondo et al (2001).…”

Section: Methodsmentioning

confidence: 99%

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Heterologous expression of laccase cDNA from Ceriporiopsis subvermispora yields copper-activated apoprotein and complex isoform patterns

et al. 2003

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Analysis of genomic clones encoding a putative laccase in homokaryon strains of Ceriporiopsis subvermispora led to the identification of an allelic variant of the previously described lcs-1 gene. A cDNA clone corresponding to this gene was expressed in Aspergillus nidulans and in Aspergillus niger. Enzyme assays and Western blots showed that both hosts secreted active laccase. Relative to the isozymic forms of the native C. subvermispora enzyme, the A. niger-produced laccase had a higher molecular mass and gave a single band on IEF gels. In contrast, A. nidulans transformants secreted several isoforms remarkably similar to those of the native system. Considered together with previously reported Southern blots and protein sequencing, expression in A. nidulans supports the view that C. subvermispora has a single laccase gene and that multiple isoforms result from post-translational processes. In addition, several lines of evidence strongly suggest that under copper limitation, A. nidulans secretes apoprotein which can be reconstituted by a short incubation with Cu(I) and to a lesser extent with Cu(II).

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Section: Discussionsupporting

confidence: 49%

“…Zymograms were as previously reported (Larrondo et al, 2001). Briefly, samples were applied with nonreducing denaturing loading buffer without boiling and subjected to SDS-PAGE at 4˚C.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…K m values for ABTS were determined with 0?01 U laccase, using Eadie-Hofstee plots. For endoglycosidase treatment, 5 mg enzyme was treated as described by Larrondo et al (2001).…”

Section: Methodsmentioning

confidence: 99%

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