Diversity of Glycosyl Hydrolases from Cellulose-Depleting Communities Enriched from Casts of Two Earthworm Species

被引:53
作者
Beloqui, Ana [1 ]
Nechitaylo, Taras Y. [2 ]
Lopez-Cortes, Nieves [1 ]
Ghazi, Azam [1 ]
Guazzaroni, Maria-Eugenia [1 ]
Polaina, Julio [3 ]
Strittmatter, Axel W. [4 ]
Reva, Oleg [5 ]
Waliczek, Agnes [2 ]
Yakimov, Michail M. [6 ]
Golyshina, Olga V. [2 ,7 ]
Ferrer, Manuel [1 ]
Golyshin, Peter N. [2 ,7 ,8 ]
机构
[1] CSIC, Inst Catalysis, E-28049 Madrid, Spain
[2] HZI Helmholtz Ctr Infect Res, D-38124 Braunschweig, Germany
[3] CSIC, Inst Agroquim & Tecnol Alimentos, Valencia 46980, Spain
[4] Eurofins MWG Operon, D-85560 Ebersberg, Germany
[5] Univ Pretoria, Dept Biochem, ZA-0002 Pretoria, South Africa
[6] CNR, Ist Ambiente Marino Costiero, I-98122 Messina, Italy
[7] Bangor Univ, Sch Biol Sci, Bangor LL57 2UW, Gwynedd, Wales
[8] Aberystwyth Univ Bangor Univ Partnership, Ctr Integrated Res Rural Environm CRRE, Aberystwyth SY23 3BF, Ceredigion, Wales
关键词
CRYSTAL-STRUCTURE; APORRECTODEA-CALIGINOSA; SUBSTRATE-SPECIFICITY; LUMBRICUS-TERRESTRIS; FUNCTIONAL-ANALYSIS; GLYCOLYTIC-ENZYMES; BACTERIAL GENOMES; FAMILY; GUT; GENE;
D O I
10.1128/AEM.00902-10
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The guts and casts of earthworms contain microbial assemblages that process large amounts of organic polymeric substrates from plant litter and soil; however, the enzymatic potential of these microbial communities remains largely unexplored. In the present work, we retrieved carbohydrate-modifying enzymes through the activity screening of metagenomic fosmid libraries from cellulose-depleting microbial communities established with the fresh casts of two earthworm species, Aporrectodea caliginosa and Lumbricus terrestris, as inocula. Eight glycosyl hydrolases (GHs) from the A. caliginosa-derived community were multidomain endo-beta-glucanases, beta-glucosidases, beta-cellobiohydrolases, beta-galactosidase, and beta-xylosidases of known GH families. In contrast, two GHs derived from the L. terrestris microbiome had no similarity to any known GHs and represented two novel families of beta-galactosidases/alpha -arabinopyranosidases. Members of these families were annotated in public databases as conserved hypothetical proteins, with one being structurally related to isomerases/dehydratases. This study provides insight into their biochemistry, domain structures, and active-site architecture. The two communities were similar in bacterial composition but significantly different with regard to their eukaryotic inhabitants. Further sequence analysis of fosmids and plasmids bearing the GH-encoding genes, along with oligonucleotide usage pattern analysis, suggested that those apparently originated from Gammaproteobacteria (pseudomonads and Cellvibrio-like organisms), Betaproteobacteria (Comamonadaceae), and Alphaproteobacteria (Rhizobiales).
引用
收藏
页码:5934 / 5946
页数:13
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