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.
Xu
Li
L
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.
Jiang
Xueqing
X
Yang
Jiangke
J
Liu
Yun
Y
Yan
Yunjun
Y
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