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Isolation and Characterization of Glucose Dehydrogenase Gene in Methylovorus sp. MP688
Author(s): 
Pages: 805-809
Year: Issue:  6
Journal: Letters in Biotechnology

Keyword:  glucose dehydrogenasemethylotrophic bacteriaexpression and purificationpyrroloquinoline quinone;
Abstract: Objective: To isolate glucose dehydrogenase genes in Methylovorus sp. MP688 and to study the func-tion of the enzyme. Methods: The protein sequence in Methylovorus sp. MP688 which shared the highest similari-ty with typical bacterial glucose dehydrogenase was isolated through BLAST program from NCBI, containing trans-membrane domain at its amino terminal. The fragment of mpq_2164 and s-mpq_2164(without the sequence of transmembrane domain) were first amplified from genome of MP688 and ligated with pET-15b. The resultant plas-mids were then transformed into E.coli BL21 respectively to acquire the recombinant proteins. The 6 × His-tagged proteins was induced by 1 mmol/L IPTG and purified by Ni2+ affinity chromatography. The glucose dehydrogenase activity was determined by using DCIP method. Results: A putative glucose dehydrogenase gene was isolated and the corresponding catalytic domain was successfully expressed in a soluble form in E.coli BL21. Although the pro-tein sequence of MPQ_2164 shared very low similarity with reported glucose dehydrogenase, it could oxidise D-glu-cose in the presence of pyrroloquinoline quinone(PQQ). Conclusion: MPQ_2164 served as a PQQ dependent glu-cose dehydrogenase in Methylovorus sp. MP688. Deletion of the transmembrane region promoted the protein expres-sion in E.coli.
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