Evidence for lignin oxidation by the giant panda fecal microbiome

PLoS One. 2012;7(11):e50312. doi: 10.1371/journal.pone.0050312. Epub 2012 Nov 28.

Abstract

The digestion of lignin and lignin-related phenolic compounds from bamboo by giant pandas has puzzled scientists because of the lack of lignin-degrading genes in the genome of the bamboo-feeding animals. We constructed a 16S rRNA gene library from the microorganisms derived from the giant panda feces to identify the possibility for the presence of potential lignin-degrading bacteria. Phylogenetic analysis showed that the phylotypes of the intestinal bacteria were affiliated with the phyla Proteobacteria (53%) and Firmicutes (47%). Two phylotypes were affiliated with the known lignin-degrading bacterium Pseudomonas putida and the mangrove forest bacteria. To test the hypothesis that microbes in the giant panda gut help degrade lignin, a metagenomic library of the intestinal bacteria was constructed and screened for clones that contained genes encoding laccase, a lignin-degrading related enzyme. A multicopper oxidase gene, designated as lac51, was identified from a metagenomic clone. Sequence analysis and copper content determination indicated that Lac51 is a laccase rather than a metallo-oxidase and may work outside its original host cell because it has a TAT-type signal peptide and a transmembrane segment at its N-terminus. Lac51 oxidizes a variety of lignin-related phenolic compounds, including syringaldazine, 2,6-dimethoxyphenol, ferulic acid, veratryl alcohol, guaiacol, and sinapinic acid at conditions that simulate the physiologic environment in giant panda intestines. Furthermore, in the presence of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), syringic acid, or ferulic acid as mediators, the oxidative ability of Lac51 on lignin was promoted. The absorbance of lignin at 445 nm decreased to 36% for ABTS, 51% for syringic acid, and 51% for ferulic acid after incubation for 10 h. Our findings demonstrate that the intestinal bacteria of giant pandas may facilitate the oxidation of lignin moieties, thereby clarifying the digestion of bamboo lignin by the animal.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Copper / metabolism
  • Escherichia coli / metabolism
  • Feces
  • Gene Library
  • Intestines / microbiology
  • Laccase / chemistry*
  • Laccase / metabolism
  • Lignin / metabolism*
  • Metagenome*
  • Metagenomics
  • Molecular Sequence Data
  • Molecular Weight
  • Oxidoreductases / metabolism
  • Oxygen / metabolism*
  • Phenol / chemistry
  • Phylogeny
  • Protein Structure, Tertiary
  • Pseudomonas / metabolism
  • RNA, Ribosomal, 16S / metabolism
  • Sasa
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Ursidae

Substances

  • RNA, Ribosomal, 16S
  • Phenol
  • Copper
  • Lignin
  • Oxidoreductases
  • Laccase
  • copper oxidase
  • Oxygen

Grants and funding

This work was supported by the Scientific Research Foundation for Returned Scholars, Ministry of Education of China (to YX); the Natural Science Foundation of Anhui Province (11040606Q05 to WF); the Natural Science Foundation of the Department of Education of Anhui Province (KJ2010A013 to WF); the Introduction Project of Academic and Technology Leaders in Anhui University (32030066 to ZF) and the Innovative Research Team Program of 211 Project in Anhui University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.