Culture independent PCR: an alternative enzyme discovery strategy.

TY - JOUR

T1 - Culture independent PCR: an alternative enzyme discovery strategy.

AU - Jacobsen, Jonas

AU - Lydolph, Magnus

AU - Lange, Lene

N1 - Keywords: Amino Acid Sequence; Ascomycota; Base Sequence; Basidiomycota; Cellulase; Cellulose 1,4-beta-Cellobiosidase; Cloning, Molecular; DNA, Fungal; Molecular Sequence Data; Phylogeny; Plant Leaves; Polymerase Chain Reaction; Sequence Alignment; Zea mays

PY - 2005

Y1 - 2005

N2 - Degenerate primers were designed for use in a culture-independent PCR screening of DNA from composite fungal communities, inhabiting residues of corn stovers and leaves. According to similarity searches and alignments amplified clone sequences affiliated with glycosyl hydrolase family 7 and glycosyl hydrolase family 45 though significant sequence divergence was observed. Glycosyl hydrolases from families 7 and 45 play a crucial role in biomass conversion to fuel ethanol. Research in this renewable energy source has two objectives: (i) To contribute to development of a renewable alternative to world's limited crude fossil oil reserves and (ii) to reduce air pollution. Amplification with 18S rDNA-specific primers revealed species within the ascomycetous orders Sordariales and Hypocreales as well as basidiomycetous order Agaricales to be present in these communities. Our study documents the value of culture-independent PCR in microbial diversity studies and could add to development of a new enzyme screening technology.

AB - Degenerate primers were designed for use in a culture-independent PCR screening of DNA from composite fungal communities, inhabiting residues of corn stovers and leaves. According to similarity searches and alignments amplified clone sequences affiliated with glycosyl hydrolase family 7 and glycosyl hydrolase family 45 though significant sequence divergence was observed. Glycosyl hydrolases from families 7 and 45 play a crucial role in biomass conversion to fuel ethanol. Research in this renewable energy source has two objectives: (i) To contribute to development of a renewable alternative to world's limited crude fossil oil reserves and (ii) to reduce air pollution. Amplification with 18S rDNA-specific primers revealed species within the ascomycetous orders Sordariales and Hypocreales as well as basidiomycetous order Agaricales to be present in these communities. Our study documents the value of culture-independent PCR in microbial diversity studies and could add to development of a new enzyme screening technology.

U2 - 10.1016/j.mimet.2004.08.013

DO - 10.1016/j.mimet.2004.08.013

M3 - Journal article

C2 - 15567226

SN - 0167-7012

VL - 60

SP - 63

EP - 71

JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

IS - 1

ER -