Cloning and expression of delta-1-pyrroline-5-carboxylate dehydrogenase in Escherichia coli DH5 alpha improves phosphate solubilization

文献类型： 外文期刊

第一作者： Gong, Mingbo

作者： Gong, Mingbo;Tang, Chaoxi;Zhu, Changxiong

作者机构：

关键词： phosphate solubilizing;Penicillium oxalicum;SMART;delta-1-pyrroline-5-carboxylate dehydrogenase;organic acid

期刊名称：CANADIAN JOURNAL OF MICROBIOLOGY （ 影响因子：2.419； 五年影响因子：2.35 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： A primary cDNA library of Penicillium oxalicum I1 was constructed using the switching mechanism at the 5' end of the RNA transcript (SMART) technique. A total of 106 clones showed halos in tricalcium phosphate (TCP) medium, and clone I-40 showed clear halos. The full-length cDNA of clone I-40 was 1355 bp with a complete open reading frame (ORF) of 1032 bp, encoding a protein of 343 amino acids. Multiple alignment analysis revealed a high degree of homology between the ORF of clone I-40 and delta-1-pyrroline-5-carboxylate dehydrogenase (P5CDH) of other fungi. The ORF expression vector was constructed and transformed into Escherichia coli DH5 alpha. The transformant (ORF-1) with the P5CDH gene secreted organic acid in medium with TCP as the sole source of phosphate. Acetic acid and alpha-ketoglutarate were secreted in 4 and 24 h, respectively. ORF-1 decreased the pH of the medium from 6.62 to 3.45 and released soluble phosphate at 0.172 mg . mL(-1) in 28 h. Expression of the P. oxalicum I1 p5cdh gene in E. coli could enhance organic acid secretion and phosphate-solubilizing ability.

分类号： Q93

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