Thin layer chromatography identification of flavoprotein compounds in crude local spinach extract

Abstract

Spinach serves as a source of a nutritious plant rich in many compounds. Components in the leaf extract of spinach are beneficial for humans. In addition to that, spinach leaves contain flavoproteins which are a class of enzymes mediating electron transport in biological system. They possess a protein chromophore with flavin adenine dinucleotide (FAD) as prosthetic group. Spinach redox proteins are used in vitro enzymatic reconstitution studies to replace native redox proteins found in bacteria. However, datathat supports the role of spinach redox proteins as control redox partners for Mycobacterium tuberculosis (Mtb) cytochrome P450s are sparse. Therefore, this study focuses on local spinach leaves containing FAD which can transport electrons. Spinach leaves extracted using gentle mechanical process with potassium phosphate buffer (pH 7.4). Paper chromatography (PC) shows positive result for flavin, thin layer chromatography (TLC) shows compound RF value similar to standard riboflavin (0.16), spectrophotometer analysis and urea denaturation test did not shows spectra of flavin compound in sample. In chemical oxidation and reduction, spinach extracts shows positive reaction towards potassium ferricyanide (oxidant) indicating electron displacement in spinach samples. Preparative chromatography methods need to be developed in order to purify crude spinach extracts to obtain semi-pure or pure flavoproteins.