Abstract
Enzyme-mediated esterification reactions can be a promising alternative to produce esters of commercial interest, replacing conventional chemical processes. The aim of this work was to verify the potential of an esterase for ester synthesis. For that, recombinant lipolytic enzyme EST5 was purified and presented higher activity at pH 7.5, 45 °C, with a Tm of 47 °C. Also, the enzyme remained at least 50% active at low temperatures and exhibited broad substrate specificity toward p-nitrophenol esters with highest activity for p-nitrophenyl valerate with a Kcat/Km of 1533 s−1 mM−1. This esterase exerted great properties that make it useful for industrial applications, since EST5 remained stable in the presence of up to 10% methanol and 20% dimethyl sulfoxide. Also, preliminary studies in esterification reactions for the synthesis of methyl butyrate led to a specific activity of 127.04 U·mg−1. The enzyme showed higher esterification activity compared to other literature results, including commercial enzymes such as LIP4 and CL of Candida rugosa assayed with butyric acid and propanol which showed esterification activity of 86.5 and 15.83 U·mg−1, respectively. In conclusion, EST5 has potential for synthesis of flavor esters, providing a concept for its application in biotechnological processes.
Highlights
BlastP analysis based on the information in the GenBank database revealed highest identity (78%) of the corresponding protein EST5 with a lipase/esterase from uncultured bacterium derived from sea sediment sample, followed by 74%
The DNA sequence of est5 gene has been deposited in GenBank with the accession number KY563703
To analyze the phylogenetic relationship of EST5 with other lipolytic enzymes, a neighbor-joining tree was constructed based on amino acid sequences of lipolytic enzymes representing the eight families proposed by Arpigny and Jaeger (1999)
Summary
The use of industrial enzymes has arisen as a significant solution for green and sustainable industrial products [4] and can be incorporated into the concept of the “circular economy” where nothing is wasted [5] In this context, lipolytic enzymes are highly useful in many industrially significant processes due to their stability in the presence of solvents; exquisite chemo-, regio- and enantioselectivities; activity over a broad range of substrates; and no need for cofactors [6,7,8,9,10]. They are promising and have high-growth potential in the valuable world industrial enzymes market; the global lipase market is expected to reach $590.5 million in 2020 [17,18]
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