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Assembly in vitro of rhodococcus jostii RHA1 encapsulin and peroxidase DypB to form a nano-compartment
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Rahmanpour, Rahman and Bugg, Tim (2013) Assembly in vitro of rhodococcus jostii RHA1 encapsulin and peroxidase DypB to form a nano-compartment. FEBS Journal, Volume 280 (Issue 9). pp. 2097-2104. doi:10.1111/febs.12234 ISSN 1742-464x.
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Official URL: http://dx.doi.org/10.1111/febs.12234
Abstract
Rhodococcus jostii RHA1 peroxidase DypB has been recently identified as a bacterial lignin peroxidase. The dypB gene is co-transcribed with a gene encoding an encapsulin protein, shown in Thermotoga maritima to assemble to form a 60-subunit nano-compartment, and DypB contains a C-terminal sequence motif thought to target the protein to the encapsulin nanocompartment. R. jostii RHA1 encapsulin protein has been overexpressed in R. jostii RHA1, and purified as a high molecular weight assembly (Mr >106). The purified nanocompartment can be denatured to form a low molecular weight species by treatment at pH 3.0, and can be re-assembled to form the nanocompartment at pH 7.0. Recombinant DypB can be assembled in vitro with monomeric encapsulin to form an assembly of similar size and shape to the encapsulin-only nanocompartment, assessed by dynamic light scattering. The assembled complex shows enhanced lignin degradation activity per mg DypB present, compared with native DypB, using a nitrated lignin UV-vis assay method. The measured stoichiometry of 8.6 µmoles encapsulin/µmol DypB in the complex is comparable to the value of 10 predicted from the crystal structure.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QD Chemistry Q Science > QH Natural history > QH426 Genetics Q Science > QR Microbiology |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | ||||
Library of Congress Subject Headings (LCSH): | Genomics, Molecular genetics , Lignin , Peroxidase, Biochemistry | ||||
Journal or Publication Title: | FEBS Journal | ||||
Publisher: | Blackwell | ||||
ISSN: | 1742-464x | ||||
Official Date: | 2013 | ||||
Dates: |
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Volume: | Volume 280 | ||||
Number: | Issue 9 | ||||
Number of Pages: | 8 | ||||
Page Range: | pp. 2097-2104 | ||||
DOI: | 10.1111/febs.12234 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access | ||||
Copyright Holders: | FEBS | ||||
Funder: | University of Warwick |
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