Article (Scientific journals)
Screening of a soil metatranscriptomic library by functional complementation of Saccharomyces cerevisiae mutants.
Kellner, Harald; Luis, Patricia; Portetelle, Daniel et al.
2011In Microbiological Research, 166 (5), p. 360–368
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Abstract :
[en] Metatranscriptomics applied to environmental transcripts provides unique opportunities to reveal microbial activity in the environment and to discover novel enzymes of potential use in biotechnological applications. Here, by functional complementation of a pho5(-) mutation (affecting a repressible acid phosphatase) and a his3(-) mutation in Saccharomyces cerevisiae, we identified fungal genes encoding an acid phosphatase and an imidazoleglycerol-phosphate dehydratase in a metatranscriptomic library, which was obtained by reverse-transcribed polyA fraction of total RNA extracted from the organic layer of a sugar maple forest soil, constructed in the modified yeast secretion vector pTEF-MF-SfiI A/B. Yeast transformants exhibiting phosphatase activity were identified in a colony-staining assay and transformants with his3(-)-complementing genes were detected by plating on histidine-deficient medium. In each screen one DNA insert was found and sequenced. The sequenced his3(-)-complementing gene showed strong similarity to a basidiomycete imidazoleglycerol-phosphate dehydratase (76% identity to a Phaffia rhodozyma enzyme). The candidate showing phosphatase activity was found to produce phosphatase extracellularly, the enzyme showing highest activity at pH 4 and between 40 and 50°C when 4-nitrophenyl phosphate was used as substrate. The sequenced insert showed strong similarity to a basidiomycete acid phosphatase (60% identity to Postia placenta).
Disciplines :
Microbiology
Author, co-author :
Kellner, Harald ;  Université de Liège - ULiège > Chimie et bio-industries > Biologie animale et microbienne
Luis, Patricia
Portetelle, Daniel ;  Université de Liège - ULiège > Chimie et bio-industries > Biologie animale et microbienne
Vandenbol, Micheline ;  Université de Liège - ULiège > Chimie et bio-industries > Biologie animale et microbienne
Language :
English
Title :
Screening of a soil metatranscriptomic library by functional complementation of Saccharomyces cerevisiae mutants.
Publication date :
2011
Journal title :
Microbiological Research
ISSN :
0944-5013
Publisher :
Elsevier Science
Volume :
166
Issue :
5
Pages :
360–368
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 28 September 2010

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