Determination of Malic Acid Using a Malate Dehydrogenase Reactor After Purification and Immobilization in Non-Denaturing Conditions and Staining with Ponceau S

Title: Determination of Malic Acid Using a Malate Dehydrogenase Reactor After Purification and Immobilization in Non-Denaturing Conditions and Staining with Ponceau S

Volume: 17 Issue: 8

Author(s): Youji Shimazaki and Takahiro Sakikawa

Affiliation:

Keywords: Enzyme reactor, immobilization, malic acid, malate dehydrogenase, Ponceau S, two-dimensional electrophoresis

Abstract: Mouse liver cytosolic malate dehydrogenase was separated by non-denaturing two-dimensional electrophoresis and identified. Furthermore, the activity of the enzyme was preserved even after separation, electroblotting onto a membrane and staining with Ponceau S in acidic buffer solution (pH 5.1). Using the membrane-immobilized enzyme, the malic acid content was estimated by measuring absorbance changes due to the conversion of nicotinamide adenine dinucleotide (NAD) to NADH. These results indicate that enzyme reactors can be systematically produced after purification, immobilization and staining with Ponceau S.

Cite this article as:

Shimazaki Youji and Sakikawa Takahiro, Determination of Malic Acid Using a Malate Dehydrogenase Reactor After Purification and Immobilization in Non-Denaturing Conditions and Staining with Ponceau S, Protein & Peptide Letters 2010; 17 (8) . https://dx.doi.org/10.2174/092986610791498867

DOI https://dx.doi.org/10.2174/092986610791498867	Print ISSN 0929-8665
Publisher Name Bentham Science Publisher	Online ISSN 1875-5305