Cloning and expression of an alpha-amylase gene from Phanerochaete chrysosporium.
Based on the genomic sequence and cDNA library screening, the cDNA sequence encoding an alpha-amylase was cloned from the filamentous white-rot fungus Phanerochaete chrysosporium and designated as pcamy1. Alignment results showed that the predicted protein has up to 43% amino acid homology to the known alpha-amylases in other organisms and is close to those from some filamentous fungi. Under nitrogen-starvation condition, the transcription of pcamy1 was accordingly upregulated or downregulated when soluble starch or glucose is sole carbon source. Addition of oxygen to nitrogen-limited media led to pcamy1 transcription and removal of glucose metabolic repression. The result indicated that the pcamy1 transcript was not only regulated by nutrients such as the carbon source but also by the cultivation environment, such as oxygen. This coordinate-regulatory model is likely common in P. chrysosporium. The expressed product of this gene in Escherichia coli could hydrolyze soluble starch, and its enzymatic activity was determined. As far as we know, this is the first report about cloning and expression study on the alpha-amylase in P. chrysosporium.
Wu
Bo
B
College of Life Sciences, Sichuan Key Laboratory of Molecular Biology and Biotechnology, Sichuan University, 29 Wangjiang Road, Chengdu, 610064, Sichuan Province, PR China.
Hu
Guo-Ku
GK
Feng
Hong
H
Wu
Jin-Ming
JM
Zhang
Yi-Zheng
YZ
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D020075:Phanerochaete / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; D004271:DNA, Fungal; D011994:Recombinant Proteins; D000516:alpha-Amylases