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Cloning of a novel gene from Penicillium oxalicum I1 which in Escherichia coli enhances the secretion of acetic acid

文献类型： 外文期刊

作者： Liu Xue 1 ; Zhu Chang-xiong 1 ; Gong Ming-bo 2 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Environm & Sustainable Dev Agr, Beijing 100081, Peoples R China

2.Chinese Acad Agr Sci, Key Lab Microbial Resources, Minist Agr, Inst Agr Resources & Reg Planning, Beijing 100081, Peoples R China

3.Univ Liege, Gembloux Agrobio Tech Microbial Proc & Interact M, Passage Deportes 2, BE-5030 Gembloux, Belgium

关键词： Penicillium oxalicum;organic acids;cloning;gene banks

期刊名称：BIOTECHNOLOGIE AGRONOMIE SOCIETE ET ENVIRONNEMENT （ 影响因子：1.087； 五年影响因子：1.523 ）

ISSN： 1370-6233

年卷期： 2018 年 22 卷 1 期

页码：

收录情况： SCI

摘要： Description of the subject. Organic acids play an important role in the conversion of insoluble ions into soluble ones in soil. Heterologous overexpression of a single gene in a cell is the optimal strategy for increasing the secretion of organic acids solubilizing phosphate. Objectives. In this study, we constructed a primary cDNA library of Penicillium oxalicum I1, and screened clones that can solubilize P in tricalcium phosphate (TCP) medium. We aimed to obtain the gene expressed in Escherichia coli, which can enhance organic acid secretion. Method. A primary cDNA library of Penicillium oxalicum I1 was constructed using the switching mechanism at the 5'-end of RNA transcription. The organic acid secretion ability of E. coli DH5 alpha (TM) with overexpressed P. oxalicum I1gene was tested in TCP medium where glucose is the sole carbon source. Afterwards, pyruvic acid, citric acid, a-ketoglutaric acid, succinic acid, fumaric acid, and malic acid were used as sole carbon source substitutes for glucose in the TCP medium to test the organic acid secretion ability of the transformed E. coli DH5 alpha (TM). Results. A total of 106 clones showed halos in TCP medium, among which clone I-2 displayed clear halo. The full-length cDNA of clone I-2 was 1,151 bp, with a complete open reading frame of 702 bp, which encoded a hypothetical protein of 233 amino acids. The cDNA sequence showed 68% identity and 73% query cover with other fungal gene sequences of which the function remains unknown. Escherichia coli containing the cloned gene secreted up to 567 mg.l(-1) acetic acid within 48 h. The use of glucose, pyruvic acid, a-ketoglutaric acid, and malic acid improved the acetic acid secretion of the E. coli DH5 alpha (TM) clone I-2. By contrast, the use of citric acid, succinic acid, and fumaric acid did not improve the acetic acid secretion of clone I-2 compared to a control E. coli DH5 alpha (TM) strain bearing only the cloning vector without any insert. Conclusions. We obtained a novel gene from Penicillium oxalicum I1 whose overexpression in E. coli DH5 alpha (TM) increased the secretion of acetic acid. This observation should help to understand what is the function of the gene isolated from P. oxalicum as well as that of its homologs found in several other species of the Penicillium genus.

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