Cloning of a novel lipase gene, lipJ08, from Candida rugosa and expression in Pichia pastoris by codon optimization.
A novel lipase gene, lipJ08, was cloned from Candida rugosa ATCC14830, along with the already reported five lipase genes (lip1-lip5). Nucleotide sequencing indicated that the lipJ08 gene contains a 1650 bp open reading frame (ORF) without introns. The deduced amino acid sequence corresponds to 534 amino acid residues, including a putative signal sequence of 15 amino acid residues. Seventeen of the non-universal serine codons (CTG) of lipJ08 were converted into universal serine codons (TCT) by PCR-based mutagenesis. The native and codon-optimized lipJ08 genes were expressed in Pichia pastoris. The hydrolytic activity of the recombinant LIPJ08 was 4.7 U/ml, whereas the activity of the recombinant wild-type lipase could not be detected.
Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, 430074 Wuhan, People's Republic of China.
Li
L
Xu
Xueqing
X
Jiang
Jiangke
J
Yang
Yun
Y
Liu
Yunjun
Y
Yan
D002175:Candida / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D003062:Codon; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D008049:Lipase / Q000235:genetics* / Q000378:metabolism*; D010843:Pichia / Q000201:enzymology* / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism
D003062:Codon; D005656:Fungal Proteins; D011994:Recombinant Proteins; C428110:Lip4 protein, Candida rugosa; D008049:Lipase