lip2, a novel lipase gene cloned from Aspergillus niger exhibits enzymatic characteristics distinct from its previously identified family member.
We have cloned a novel lipase gene, lip2, from Aspergillus niger and expressed it in Escherichia coli. Upon purification of the recombinant Lip2 protein, its properties were characterized. In comparison with a previously identified lipase Lip1, both enzymes are acid lipases (optimal pH <6.5), Ca(2+)-dependent and PMSF-sensitive, but have different molecular weights (35 and 43 kDa), optimal substrate spectra (C10 and C8), optimal reaction temperatures (45 and 50 degrees C) and thermal stability. Circular dichroism spectroscopy revealed that Lip2 contains a typical Ca(2+)-active site. This first report on the cloning of the Lip2 gene and its enzymatic characteristics may greatly facilitate its potential industrial application.
Yang
Jiangke
J
Key Laboratory of Molecular Bio-Physics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, 430074, Wuhan, China. jiangke.yang@gmail.com
Sun
Jin
J
Yan
Yunjun
Y
D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D002118:Calcium / Q000494:pharmacology; D020134:Catalytic Domain; D002413:Cations, Divalent / Q000494:pharmacology; D002942:Circular Dichroism; D003001:Cloning, Molecular; D003067:Coenzymes / Q000494:pharmacology; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004791:Enzyme Inhibitors; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity
D002413:Cations, Divalent; D003067:Coenzymes; D004271:DNA, Fungal; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D010664:Phenylmethylsulfonyl Fluoride; D008049:Lipase; D002118:Calcium