Cloning of an acidic laccase gene (clac2) from Coprinus congregatus and its expression by external pH

Abstract Coprinus congregatus synthesized and secreted high amounts of laccase under acidic culture condition (pH 4.1) transiently. We have cloned a genomic DNA fragment between the copper binding regions I and II of a laccase by PCR from C. congregatus. This fragment was used as a probe to clone a...

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Bibliographic Details
Published in:FEMS microbiology letters 2001-02, Vol.195 (2), p.151-156
Main Authors: Kim, Soonja, Leem, Youngeun, Kim, Kyunghoon, Choi, Hyoung T.
Format: Article
Language:English
Subjects:
Acidic stress
Amino Acid Sequence
Amino acids
Base Sequence
Binding
Biological and medical sciences
Blotting, Northern
cDNA cloning
clac2 gene
Cloning
Cloning, Molecular
Copper
Coprinus - enzymology
Coprinus - genetics
Coprinus congregatus
Culture
Deoxyribonucleic acid
DNA
DNA, Complementary
Fundamental and applied biological sciences. Psychology
Fungal Proteins
Gene expression
Gene Expression Regulation, Fungal
Genes, Fungal
Growth, nutrition, metabolism, transports, enzymes. Molecular biology
Hydrogen-Ion Concentration
Laccase
Laccase expression
Microbiology
Molecular Sequence Data
Mycology
Northern blotting
Oxidoreductases - chemistry
Oxidoreductases - genetics
Oxidoreductases - metabolism
pH effects
Polymerase Chain Reaction
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Summary:Abstract Coprinus congregatus synthesized and secreted high amounts of laccase under acidic culture condition (pH 4.1) transiently. We have cloned a genomic DNA fragment between the copper binding regions I and II of a laccase by PCR from C. congregatus. This fragment was used as a probe to clone a laccase cDNA from the acidic culture. The cDNA (clac2) consisted of 1817 bp which contains 1086 bp encoding 362 amino acids. The N-terminal sequence of the purified CLAC2 revealed that the first 16 amino acids were removed by the modification. The purified protein had two copper binding regions, although other fungal laccases had four. According to a Northern blotting analysis the clac2 was expressed not in neutral culture but in acidic culture only. The transcription of the clac2 as well as the laccase activity reached a maximum after 24 h upon transferring to an acidic culture, and then decreased very rapidly.
ISSN:0378-1097
1574-6968
DOI:10.1111/j.1574-6968.2001.tb10513.x