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. 2021 Dec;36(1):248-256.
doi: 10.1080/14756366.2020.1861607.

Production, purification and biochemical characterisation of a novel lipase from a newly identified lipolytic bacterium Staphylococcus caprae NCU S6

Affiliations

Production, purification and biochemical characterisation of a novel lipase from a newly identified lipolytic bacterium Staphylococcus caprae NCU S6

Junxin Zhao et al. J Enzyme Inhib Med Chem. 2021 Dec.

Abstract

A novel lipase, SCNL, was isolated from Staphylococcus caprae NCU S6 strain in the study. The lipase was purified to homogeneity with a yield of 6.13% and specific activity of 502.76 U/mg, and its molecular weight was determined to be approximately 87 kDa. SCNL maintained above 80% of its initial activity at a wide range of temperatures (20-50 °C) and pH values (6-11), with an optimal temperature at 40 °C and optimal pH at 9.0 with p-nitrophenyl palmitate as a substrate. SCNL exhibited a higher residual activity than the other staphylococcal lipases in the presence of common enzyme inhibitors and commercial detergents. The lipase activity was enhanced by organic solvents (isooctane, glycerol, DMSO and methanol) and metal ions (Na+, Ba2+, Ca2+, and Mn2+). The Km and Vmax values of SCNL were 0.695 mM and 262.66 s-1 mM-1, respectively. The enzyme showed a preference for p-NP stearate, tributyrin and canola oil. These biochemical features of SCNL suggested that it may be an excellent novel lipase candidate for industrial and biotechnological applications.

Keywords: Staphylococcus caprae; biochemical characterisation; lipase; purification.

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Conflict of interest statement

No potential conflict of interest was reported by the author(s).

Figures

Figure 1.
Figure 1.
A phylogenetic tree displaying the relationship between strain NCU S6 and other Staphylococcus species based on partial 16S rDNA sequences of the strains of the genus. The tree was constructed using a neighbour joining algorithm in MEGA 7.0 software and the accession numbers are shown in parenthesis.
Figure 2.
Figure 2.
(A) Results of coagulase tube test, novobiocin (5 μg/piece) sensitivity test, and catalase production test. (B) The result of 16S rDNA agarose gel electrophoresis analysis. (C) The image of Gram-staining result of S. caprae NCU S6.
Figure 3.
Figure 3.
The time courses of cell growth (▲) of S. caprae NCU S6 and lipase production (■). The culture was carried out at 37 °C and 200 rpm for 111 h and cell growth was monitored by measuring the absorbance at 600 nm every 3 h.
Figure 4.
Figure 4.
SDS–PAGE electrophoretogram of the lipase from different purification stages. Lane 1: crude enzyme, Lane 2: purified enzyme from ion-exchange chromatography, Lane 3: final purified enzyme from gel filtration chromatography.
Figure 5.
Figure 5.
Effect of temperature (A) and temperature-time (B), pH (C) and pH-time (D) on activity and stability of the lipase.

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