Monascus anka Pyruvate Decarboxylase Accounts for the New Candidate Resources of Fuel Ethanol Production

Abstract:

In order to study the nature and function of Pyruvate decarboxylase (PDC, E.C.4.1.1.1), which is the key enzyme to produce ethanol by fermentation; full-length cDNA library was constructed with SMART technique from Monascus anka CICC 5031. The pdc gene, including a 1713-bp open reading frame, encoding a 570 amino acid protein, was obtained by screening the constructed M. anka cDNA library. The pdc gene was successfully heterologously expressed in E.coli BL21(DE3), accounting for 32.7% of total cellular proteins. Recombinant PDC was expressed in prokaryotic cells and purified by affinity chromatography, and native PDC was extracted and purified from M. anka through Sephadex G-25 and DEAE-anion exchange resin. The enzymatic characterization of both recombinant and native PDC were studied, respectively. The specific activity of recombinant and native PDC was 20.2 and 30.11U/mg respectively. Kinetic analysis indicated that recombinant and native PDC had the same optimum conditions: pH6.0, 30°C, the Km value for pyruvate of recombinant PDC was 2.6 mmol/L and native PDC was 0.56 mmol/L. The high activity and stable PDC from M. anka accounts for the new candidate resources of fuel ethanol production.

• Periodical: Advanced Materials Research (Volumes 512-515)
• Pages: 432-438
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.512-515.432
• Online since: May 2012
• Authors: Bi Yun Zhu, Lan Gao, Hao Ming Li
• Keywords: Fuel Ethanol, Monascus anka, Pyruvate Decarboxylase

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