Journal
MICROBIOLOGYOPEN
Volume 7, Issue 5, Pages -Publisher
WILEY
DOI: 10.1002/mbo3.601
Keywords
alkalistable; deep-sea hydrothermal vents; esterase; metagenomic sequence-based strategy; salt-tolerant
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Funding
- National Natural Science Foundation of China [41530967]
- China Ocean Mineral Resources RD Association [DY125-22-04]
- Natural Science Foundation of Fujian Province [2016J05074]
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A novel esterase gene selected from metagenomic sequences of deep-sea hydrothermal vents was successfully expressed in Escherichiacoli. The recombinant protein (est-OKK), which belongs to the lipolytic enzyme family V, exhibited high activity toward pNP-esters with short acyl chains and especially p-nitrophenyl butyrate. Site-mutagenesis results confirmed that est-OKK contains the nonclassical catalytic tetrad predicted by alignment and computational modeling. The est-OKK protein is a moderately thermophilic enzyme that is relatively thermostable, and highly salt-tolerant, which remained stable in 3 mol/L NaCl for 6 hr. The est-OKK protein showed the considerable alkalistability, displayed optimal activity at pH 9.0 and maintained approximately 70% of its residual activity after incubation at pH 10 for 4 hr. Furthermore, the est-OKK activity was strongly resistant to a variety of metal ions such as Co2+, Zn2+, Fe2+, Na+, and K+; nonionic detergents such as Tween-20, Tween-80; and organic solvents such as acetone and isopropanol. Taken together, the novel esterase with unique characteristics may give us a new insight into the family V of lipolytic enzymes, and could be a highly valuable candidate for biotechnological applications such as organic synthesis reactions or food and pharmaceutical industries.
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