Cloning of a novel lipase gene, <Emphasis Type="Italic">lipJ08,</Emphasis> from <Emphasis Type="Italic">Candida rugosa</Emphasis> and expression in <Emphasis Type="Italic">Pichia pastoris</Emphasis> by codon optimization |
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Authors: | Li Xu Xueqing Jiang Jiangke Yang Yun Liu Yunjun Yan |
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Institution: | (1) 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; |
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Abstract: | 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. |
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