Purification and Characterization of Active Aggregates of an Organic Solvent Tolerant Lipase from Marinobacter sp. EMB5

Abstract

Background: Marinobacter is a halophilic bacterial genus and a bio-geochemically important group in sea water, is generally associated with the bioremediation of hydrocarbon-rich effluents released into the salt-rich environment by various anthropogenic activities. It is a well-known lipase producer-the NCBI Protein database shows nearly 520 Marinobacter lipases. In this context, its lipase and esterase are considered important. However, their enzymatic properties in purified form have been scantily studied. Since hydrocarbons are acted upon on solvent water interface, the stability and catalytic behavior of Marinobacter lipase in non-aqueous media needs to be more specifically investigated.Aim: Marinobacter sp. EMB5 was isolated from Kozhikode, Kerala, India. It was found to be potent lipase producer. The aim of the present work was to obtain a purified preparation and undertake its detailed characterization with respect to above aspects. Work:The crude Marinobacter sp. lipase was purified by Sephacryl S200 gel filtration chromatography. The lipase eluted as a single activity peak in void volume and had a very high molecular mass. Aggregation has not been reported in case of Marinobacter lipase, so far, although it is known in other lipases. The aggregated lipase was mixed with 2-propanol (70%, v/v) and disaggregated lipase was obtained as a single activity peak in Sephacryl S200 gel filtration chromatography corresponding to 82 kDa molecular mass. This monomeric species was less active than aggregated purified preparation. Hence, further characterization and studies were carried out on purified active aggregate lipase. It was stable in a wide range of organic solvents at high concentration up to 24 h. The organic solvent stability was further investigated by Far UV CD spectroscopy and fluorescence spectroscopy. Conclusion:Due to the fact that Marinobacter sp. EMB5 lipase remained stable in organic solvents for prolonged incubation periods, it has potential to be used in industrial organic synthesis.

Enhanced halophilic lipase secretion by Marinobacter litoralis SW‐45 and its potential fatty acid esters release

“…The influence of surfactants or detergents on crude MLL activity is shown in Supporting Information was noticed, while increment in the concentration of SDS (from 0.1 to 0.5%) and CTAB (from 1 to 10 mM) inhibited the lipase activity. The result obtained in this study is consistent with the findings of Hemamalini and Khare [19] where they reported extreme stability of Marinobacter sp. EMB5 lipase in CTAB, Triton X, and Tween 80, and significant inhibition of lipase activity by SDS.…”

Lipase‐catalyzed hydrolysis of (+,‐)‐2‐(4‐methylphenyl) propionic methyl ester enhanced by hydroxypropyl‐β‐cyclodextrin

“…The possible reason is that a free enzyme is used in this paper and the reaction medium contains 0.1 mol L −1 phosphate buffer. In such a reaction system, there is an intense tendency to aggregate for the enzyme . The decrease of E with the concentration of CRL may be the factor that causes more enzyme molecules to aggregate with each other at a higher concentration.…”

“…In such a reaction system, there is an intense tendency to aggregate for the enzyme. [49][50][51][52][53][54] The decrease of E with the concentration of CRL may be the factor that causes more enzyme molecules to aggregate with each other at a higher concentration. It is also observed from Fig.…”

Synthesis of flavor esters by a novel lipase from Aspergillus niger in a soybean-solvent system

“…SYBC WU-3 (pH 9.5) (Cai et al 2009) and Marinobacter sp. EMB5 (pH 9.0) (Hemamalini and Khare 2016). The enzyme showed excellent stability over a wide pH range from acidic to alkaline (Fig.…”

“…SYBC WU-3 (Cai et al 2009), but lower than those from Bacillus methylotrophicus PS3 (55 °C) (Sharma et al 2017), Marinobacter sp. EMB5 (50 °C) (Hemamalini and Khare 2016) and Pichia lynferdii Y-7723 (35 °C) (Kim et al 2010). Few microbial enzymes have catalytic activity at such low temperatures (Kim et al 2010), suggesting that An-lipase might be of value in processes requiring low temperature, such as cheese ripening and alcoholic beverage production (Sumby et al 2009).…”