-transparent.png){kind=logo}
The gene encoding an alkaline lipase of Penicillium cyclopium PG37 was cloned with four steps of PCR amplification based on different principles. The cloned gene was 1,480 nucleotides in length, consisted of 94 bp of promoter region, and had 6 exons and 5 short introns ranging from 50 to 70 nucleotides. The open reading frame encoded a protein of 285 amino acid residues consisting of a 27-AA signal peptide and a 258-AA mature peptide, with a conserved motif of Gly-X-Ser-X-Gly shared by all types of alkaline lipases. However, this protein had a low homology with lipases of P. camembertii (22.9%), Humicola lanuginosa (25.6%), and Rhizomucor miehei (22.3%) at the amino acid level. The mature peptide-encoding cDNA was cloned and expressed in Escherichia coli on pET-30a for confirmation. A distinct band with a M.W. of 33 kDa was detected on SDS-PAGE. Results of a Western blot analysis and an enzyme activity assay verified the recombinant 33-kDa protein as an alkaline lipase. Its catalytic properties were not changed when compared with its natural counterpart.
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
January 2011,
Prikladnaia biokhimiia i mikrobiologiia,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
January 1999,
Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
January 1990,
FEMS microbiology letters,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
November 1992,
Current microbiology,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
April 1998,
Lipids,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
August 2000,
FEMS microbiology letters,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
July 1992,
Biochimica et biophysica acta,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
December 1997,
Wei sheng wu xue bao = Acta microbiologica Sinica,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
November 1987,
Applied and environmental microbiology,
Minchen Wu, and
Zhikang Qian, and
Peihong Jiang, and
Taishan Min, and
Chongrong Sun, and
Weida Huang
December 2002,
Applied microbiology and biotechnology,
