Abstract
Bacterial lipases with activity spanning over a broad temperature and substrate range have several industrial applications. An efficient enzyme-producing bacterium Chryseobacterium polytrichastri ERMR1:04, previously reported from Sikkim Himalaya, was explored for purification and characterization of cold-adapted lipase. Optimum lipase production was observed in 1% (v/v) rice bran oil, pH 7 at 20°C. Size exclusion and hydrophobic interaction chromatography purified the enzyme up to 21.3-fold predicting it to be a hexameric protein of 250 kDa, with 39.8 kDa monomeric unit. MALDI-TOF-MS analysis of the purified lipase showed maximum similarity with alpha/beta hydrolase (lipase superfamily). Biochemical characterization of the purified enzyme revealed optimum pH (8.0), temperature (37°C) and activity over a temperature range of 5–65°C. The tested metals (except Cu2+ and Fe2+) enhanced the enzyme activity and it was tolerant to 5% (v/v) methanol and isopropanol. The Km and Vmax values were determined as 0.104 mM and 3.58 U/mg, respectively for p-nitrophenyl palmitate. Bioinformatics analysis also supported in vitro findings by predicting enzyme's broad temperature and substrate specificity. The compatibility of the purified lipase with regular commercial detergents, coupled with its versatile temperature and substrate range, renders the given enzyme a promising biocatalyst for potential detergent formulations.
Highlights
Lipases include an important group of biocatalysts that belong to the class of triacylglycerol hydrolases (EC: 3.1.1.3) (Gupta et al, 2004)
Maximum lipase activity was observed in 1% (v/v) rice bran oil followed by olive oil
Optimum lipase production was observed at pH 7 when the bacterial isolate was incubated at 20◦C and 150 rpm for 55 h (Figure 1)
Summary
Lipases include an important group of biocatalysts that belong to the class of triacylglycerol hydrolases (EC: 3.1.1.3) (Gupta et al, 2004) They break down fats and are highly active against water-insoluble substrates. Lipases from high altitude bacteria are of significance due to the host of specific structural features that these enzymes have evolved as a mode of adaptation to frequent freezing and thawing conditions. They find potential industrial applications in diverse fields of detergent additives, food processing, environmental bioremediation in cold, and many more (Maiangwa et al, 2015). Low-temperature active lipases are greatly advantageous as detergent additives for cold washing that would diminish energy consumption as well as wear and tear of delicate fabrics (Joseph et al, 2008)
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