| 论文题目: | Molecular cloning and heterologous expression of an acid stable xylanase gene from Alternaria sp HB186 |
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| 作者: | Mao, LW; Meng, P; Zhou, C; Ma, LX; Zhang, GM; Ma, YH |
| 联系作者: | Zhang, GM |
| 刊物名称: | WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY |
| 期: | 3 |
| 卷: | 28 |
| 页: | 777-784 |
| 年份: | 2012 |
| 影响因子: | 1.532 |
| 论文下载: | 下载地址 |
| 摘要: | A new xylanase gene, named xyn186, was cloned by the genome-walking PCR method from the Alternaria sp. HB186. The sequence of xyn186 contains a 748 bp open reading frame separated by one intron with the size of 52 bp. The cDNA was obtained by DpnI-mediated intron deletion. The cDNA was cloned into pHBM905A and transformed into Pichia pastoris GS115 to screen xylanase-secreting transformants on RBB-xylan plates. The molecular mass of the enzyme was estimated to be 23 kDa on SDS-PAGE. The optimal pH and temperature of the purified enzyme is 6 and 50A degrees C, respectively. The K (m) and V (max) valued for birchwood xylan are 1.404 mg ml(-1) and 0.2748 mmol min(-1) mg(-1), respectively. The inhibitory effects of various metal ions were investigated, Cu2+ and Hg2+ ions inhibited most of the enzyme activity. The gene copy number of xyn186 in the genome of P. pastoris was estimated as two by the Real-time PCR. To date, xyn186 gene is the first xylanase gene cloned from the genus Alternaria. |
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