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产植酸酶菌株的筛选及其植酸酶基因的克隆
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辽宁省科技攻关项目资助(No. 2001101001)


Screening of Phytase-producing Strain and Cloning of Phytase Gene from the Selected Strain
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    摘要:

    从土壤中筛选到产植酸酶活性较高的烟曲霉菌株WY-2,其植酸酶最适pH为5.5,最适温度为55 ℃。通过对烟曲霉WY-2植酸酶基因进行PCR扩增,获得了一个1.5 kb大小的特异性产物,将其克隆到载体pMD18-T中。测序结果分析表明,该基因片段含有植酸酶基因完整的阅读框架 (ORF),基因全长1459 bp,其中包含一个61 bp的内含子,编码465个氨基酸,有7个潜在的糖基化位点,5′端有一编码26个氨基酸的信号肽序列。该基因与已报道的烟曲霉ATCC34625植酸酶基因有91%同源性,编码的氨基酸序列同源性为91%。

    Abstract:

    A strain with relative higher phytase-producing ability, Aspergillus fumigatus WY-2 was screened from soil. The optimal pH and temperature for activity of the phytase from A.fumigatus WY-2 were 5.5 and 55 ℃, respectively. The gene encoding the phytase was amplified from genomic DNA of the strain by PCR, and a 1.5 kb DNA fragment was obtained and then was cloned into vector pMD18-T. The sequencing analysis revealed that the DNA fragment contained a whole open reading frame (ORF) of phytase gene. The phytase gene was 1459 bp in length included with a 61 bp intron and encoded 465 amino acids. A signal peptide encoding 26 amino acids was found at 5′end of the gene. There were 7 potential glycosylation sites in the phytase. The present phytase showed 91% identity in nucleotide sequence and 91% identity in deduced amino acids sequence to the previously reported A.fumigatus ATCC34625 phytase.

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王严,高晓蓉,苏乔,王静云,安利佳. 产植酸酶菌株的筛选及其植酸酶基因的克隆[J]. 微生物学通报, 2007, 34(4): 0719-0722

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