DNA marker utilization for the sustainable production of trehalose

Abstract

Abstract Trehalose is a type of sugar that is known by its stability and resilience towards acid and low temperature. Furthermore, trehalose has numerous health benefits and has been used by several industries, including food, cosmetics, and pharmaceuticals. Even though trehalose could be easily produced using trehalose synthase (TreS) enzyme, a sustainable production of trehalose is still a problem. Our work aims to develop an approach to identify a novel trehalose synthase enzyme from various organisms, especially thermophilic bacteria, by implementing a deoxyribonucleic acid (DNA) marker technique. We first collected protein and DNA sequences from public biological databases and subsequently conducted sequence analysis. We then designed degenerate primers based on the conserved regions identified from the sequence analysis. The designed primers were subjected to primer characterization using Oligo Calc software. The primers were further validated via in-silico PCR amplification. In general, our designed primers possess the properties to work optimally. In addition, agarose gel electrophoresis that the primers successfully amplified nucleotides encoding TreS enzyme from all samples. Our findings may serve as a basis to discover the TreS enzyme variants which possess superior attributes, allowing the sustainable production of trehalose.