Developing a DNA Marker Approach for the Sustainable Production of D-Tagatose

Abstract

D-tagatose is known as a type of sugar that has low-calorie and numerous benefits. The sugar is also known to have potential for the food industry. D-tagatose can be produced biologically using the L- arabinose isomerase (L-AI) enzyme. However, sustainable production of D-Tagatose still faces an issue due to the specificity of the enzyme and the requirement of a high temperature for large-scale production. This study aims to develop an approach to discovering new bacteria that have the L-AI enzyme by implementing the DNA marker technique. We collected protein sequences from a public biological database and performed a multiple-sequence alignment. Then, the degenerate primers were designed based on the aligned sequence. The primer characterization was carried out using Oligo Calc. In-silico PCR amplification was also performed to test the primers’ specificity. Overall, the primers’ properties have met the criteria for optimally working primers. In addition, gel electrophoresis confirmed the successful amplification of the L- AI enzyme from several bacteria. Our study could be used to discover the L-AI enzyme that has the desired characteristics, which allows the sustainable production of D-tagatose.