Abstract
The current study isolated and characterized the Lip3F9 polypeptide sequence of Deschampsia antarctica Desv. (GeneBank Accession Number JX846628), which was found to be comprised of 291 base pairs and was, moreover, expressed in Pichia pastoris X-33 cells. The enzyme was secreted after 24 h of P. pastoris culture incubation and through induction with methanol. The expressed protein showed maximum lipase activity (35 U/L) with an optimal temperature of 37 °C. The lipase-expressed enzyme lost 50% of its specific activity at 25 °C, a behavior characteristic of a psychrotolerant enzyme. Recombinant enzyme activity was measured in the presence of ionic and non-ionic detergents, and a decrease in enzyme activity was detected for all concentrations of ionic and non-ionic detergents assessed.
Highlights
The Antarctic continent is considered one of the harshest ecosystems in the world [1]
The GDSL motif enzyme is a relatively newly discovered lipase, share five blocks of highly conserved homology, which are important for their classification and the active-site Ser is located close to the N-terminus [8], are widely exist in both microbe and plant species that participate in the hydrolysis and synthesis of lipids or esters [9]
NP_001054716.2, a sequence identified as a putative GDSL lipase/acylhydrolase of Orysa sativa, where the percent of sequence identity was 56/101 (56%) and the percent of positive substitutions was 70/101 (70%)
Summary
The Antarctic continent is considered one of the harshest ecosystems in the world [1]. Information about the extraction or expression of enzymes from D. antarctica is nonexistent, and only one gene has been isolated and cloned in the pGapαA expression vector of P. pastoris This protein showed increased activity when the plant was exposed to a temperature of 4 °C and UV radiation [10,11]. The identification and characterization of genes coding for lipase-like proteins in plant species tolerant to low temperatures, such as D. antarctica, can provide novel enzymes for industrial applications. To this end, this study is the first report to isolate and characterize a polypeptide with lipase activity from D. antarctica through expression in the pPICZαB Pichia pastoris vector
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