Cellulose complementing factor (Ccp) is a new member of the cellulose synthase complex (terminal complex) in Acetobacter xylinum

Naoki Sunagawa, Takaaki Fujiwara, Takanori Yoda, Shin Kawano, Yasuharu Satoh, Min Yao, Kenji Tajima, Tohru Dairi

Research output: Contribution to journalArticlepeer-review

44 Citations (Scopus)

Abstract

The cellulose complementing factor (Ccp) is known to be involved in cellulose production in the Acetobacter species. However, its precise functions remain unclear. In the current study, we identified the coding region of the ccpAx gene (ccp gene from Acetobacter xylinum) and the localization of the CcpAx in cells by generating fusion proteins tagged to an enhanced green fluorescent protein (EGFP). From the results of N-terminal sequencing of CcpAx-EGFP-fusion protein, which recovered 65% of cellulose-producing abilities of the wild-type to the ccpAx gene-knockout mutant, the ccpAx gene was determined to encode a protein with the molecular weight of 8 kDa. The amino acid sequence deduced had high similarities with the C-terminal regions of Ccp proteins from other Acetobacter species. Fluorescence microscopy analysis showed that CcpAx was longitudinally localized along with one side of the cell membrane. Additionally, the localization of AxCeSD, which is thought to be a member of the cellulose synthase complex [terminal complex (TC)] in A. xylinum, was determined in the same manner as CcpAx. Fluorescence microscopy analysis showed that AxCeSD had a localization pattern similar to that of CcpAx. Pulldown assays and isothermal titration calorimetry analysis clearly showed a significant interaction between CcpAx and AxCeSD. Taken together, these data strongly suggest that CcpAx functions as a member of the TC in A. xylinum.

Original languageEnglish
Pages (from-to)607-612
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume115
Issue number6
DOIs
Publication statusPublished - 2013 Jun
Externally publishedYes

Keywords

  • AxCeSD
  • Bacterial cellulose
  • BcsD
  • Cellulose synthetic mechanism
  • Gluconacetobacter
  • ORF2

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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