Abstract
5′-Phosphodiesterase (5′-PDE) catalyzes the hydrolysis of ribonucleic acid to obtain a mixture of ribonucleotides, such as 5′-guanosine monophosphate and 5′-adenosine monophosphate. In this study, a 5'-PDE was newly isolated and purified from Aspergillus fumigatus. Following purification, this enzyme exhibited a specific activity of 1036.76 U/mg protein, a molecular weight of 9.5 kDa, and an optimal temperature and pH for enzyme activity of 60°C and 5.0, respectively. However, its activity was partially inhibited by Fe3+, Cu2+, and Zn2+, but slightly improved by the presence of K+ and Na+. Additionally, chemical-modification experiments were also applied to investigate the structural information of 5'-PDE, in which the residues containing carboxyl and imidazole groups were essential for enzyme activity based on their localization in the 5′-PDE active site. Furthermore, purified 5′-PDE could specifically catalyze the synthesis of ribonucleotides with a Vmax 0.71 mmol/mg·min and a KM of 13.60 mg/mL.
Highlights
Interest in 50 nucleotides has increased due to their applications in food and the pharmaceutical industries [1]
Phylogenetic experiments demonstrated that strain XD-9 belonged to A. fumigatus, which was subsequently named A. fumigatus XD-9
Our results showed that the relationship between the logarithm of relative enzyme activity and time (y = 0.6222x + 0.3105) suggested that one carboxyl group located in the 50-PDE active site (Fig 2B and 2C)
Summary
Interest in 50 nucleotides has increased due to their applications in food and the pharmaceutical industries [1]. 50 nucleotides are capable of effectively improving the taste of food and exhibit synergistic effects with monosodium glutamate [2]. There are currently three methods for producing nucleotides, including microbial fermentation, chemical synthesis, and enzymatic methods [4]. Use of enzymes has incomparable advantages, such as low cost, low complexity, and high yield, making their use the most economical method in the industry [5]. It is important to obtain an excellent enzyme capable of producing 50 nucleotides
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