Detergent compatible alkaline lipase produced by marine Bacillus smithii BTMS 11

Abstract

Bacillus smithii BTMS 11, isolated from marine sediment, produced alkaline and thermostable lipase. The enzyme was purified to homogeneity by ammonium sulfate precipitation and ion exchange chromatography which resulted in 0.51 % final yield and a 4.33 fold of purification. The purified enzyme was found to have a specific activity of 360 IU/mg protein. SDS-PAGE analyses, under non-reducing and reducing conditions, yielded a single band of 45 kDa indicating the single polypeptide nature of the enzyme and zymogram analysis using methylumbelliferyl butyrate as substrate confirmed the lipolytic activity of the protein band. The enzyme was found to have 50°C and pH 8.0 as optimum conditions for maximal activity. However, the enzyme was active over wide range of temperatures (30-80°C) and pH (7.0-10.0). Effect of a number of metal salts, solvents, surfactants, and other typical enzyme inhibitors on lipase activity was studied to determine the novel characteristics of the enzyme. More than 90 % of the enzyme activity was observed even after 3 h of incubation in the presence of commercial detergents Surf, Sunlight, Ariel, Henko, Tide and Ujala indicating the detergent compatibility of B. smithii lipase. The enzyme was also found to be efficient in stain removal from cotton cloths. Further it was observed that the enzyme could catalyse ester synthesis between fatty acids of varying carbon chain lengths and methanol with high preference for medium to long chain fatty acids showing 70 % of esterification. Results of the study indicated scope for application of this marine bacterial lipase in various industries.

Biochemistry of lipolytic enzymes secreted byPenicillium solitumandCladosporium cladosporioides

“…Evidence of an additional lipase secreted by P. solitum is noteworthy, because it supports the suggestion that penicillia are capable of producing more than one extracellular lipase. 37,38) Microbial lipases can act in a wide range of pH, even though the pH optimum for bacterial enzymes is usually neutral [39][40][41] or alkaline, 8,[42][43][44][45][46] and the pH optimum of fungal enzymes is in a range 4.0-8.0. 3) Maximal hydrolytic activity also depends on the type of ester used as substrate.…”

Biochemical properties of the alkaline lipase of Bacillus flexus XJU-1 and its detergent compatibility

“…Similarly, the pH 10.0 was optimum for the alkaline lipases purified from various Bacillus species (Lesuisse et al 1993;Bora & Kalita 2008;Bora & Bora 2012). pH 8.0 and 9.0 were also optima for lipases of Bacillus species (Sharma et al 2002;Chakraborty & Raj 2008; Anna- Shariff et al 2011;Lailaja & Chandrasekaran 2013). An important lipolytic activity was seen in the alkaline region from pH 7.0 to 13.0 (Fig.…”

“…Many lipases from Bacillus species have been purified to homogeneity (Lesuisse et al 1993;Sharma et al 2002;Kambourova et al 2003;Chakraborty & Raj 2008;Annamalai et al 2011;Shariff et al 2011;Bora & Bora 2012;Kumar et al 2012;Sharma & Kanwar 2012;Lailaja & Chandrasekaran 2013). Gupta et al (2004) reported that the purification must be efficient, fast, allowing the continuous recovery of a product and cost effective.…”

“…acetone), the hydrophobic interaction chromatography is often used to totally purify the lipases since they possess a large hydrophobic region around the active site (Kordel et al 1991;Sharma & Kanwar 2012). However, other methods, such as gel filtration and ion exchange chromatography are also in use (Chakraborty & Raj 2008;Annamalai et al 2011;Bora & Bora 2012;Kumar et al 2012;Lailaja & Chandrasekaran 2013). The lipases used in the commerce are often purified from bacteria and fungi as these microbial lipases have a better stability than plant or animal lipases (Ghosh et al 1996).…”

“…There is thus a continuous demand of lipases with better properties destined for use in detergent formulation. Although many lipases of Bacillus species have been characterized (Sugihara et al 1991;Sharma et al 2002;Kambourova et al 2003;Chakraborty & Raj 2008;Annamalai et al 2011;Bora & Bora 2012;Kumar et al 2012;Sharma & Kanwar 2012;Lailaja & Chandrasekaran 2013), the alkaline lipase of Bacillus flexus has never been characterized and used in detergent preparations. The aims of the present study were to study the properties of the alkaline lipase of Bacillus flexus and its utilization as detergent additive.…”