Optimization of extracellular thermophilic highly alkaline lipase from thermophilic Bacillus sp isolated from Hotspring of Arunachal Pradesh, India

Bora, Limpon; Bora, Minakshi

doi:10.1590/s1517-83822012000100004

Cited by 76 publications

(67 citation statements)

References 35 publications

(36 reference statements)

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“…Thermophilic Bacillus sp. have previously shown the lipase production at 50-60ºC (Kim et al, 2000;Bora and Bora, 2012). However, the reports on lipase production by Bacillus sp.…”

Section: Resultsmentioning

confidence: 93%

“…Lipases being third largest selling group of enzymes with more than 1000 t of annual production and approaching to a billion dollar business. Lipases active at alkaline pH are of great importance in detergent industry where these enzymes are used for removal of fat stains and dirt (Bora and Bora, 2012). The use of alkaline lipases in detergent formulation substantially acts as a substitute for the phosphate builders in chemical detergent which are considered as major pollutant from detergent industry (Jellouli et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

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Optimization of Lipase Production by Thermo-Alkalophilic Bacillus sp. 8C

Bhosale¹,

Uzma²,

Bismile³

2015

Research J. of Microbiology

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Microbial lipases catalyzing the hydrolysis and synthesis of esters from glycerol and fatty acids are commercially important in food, dairy, detergent and leather industries. In the present study, 15 thermo-alkalophilic bacterial strains isolated from a compost soil sample were screened for lipase production in beef extract nitrate medium containing 1% tributyrin. The strain 8C, giving maximum lipolytic activity was morphologically and biochemically identified as Bacillus sp. Lipase production from strain 8C was optimized by studying effects of incubation period, initial pH of medium, carbon and nitrogen sources, metals, substrates and thiols under static conditions. Lipase production was maximum after 7 days of incubation at 50°C and pH 9.0 and when sesame oil was used as a substrate. Supplementation of ribose or raffinose and ammonium sulphate (1%) as sole sources of carbon and nitrogen also stimulated lipase production over control. Of the different thiols tested, methionine greatly increased lipase production (175 Eu mLG 1 ) whereas, FeCl 3 (50 mM) found to be most suitable metal salt for optimal lipase production (300 Eu mLG 1 ). Under optimized conditions, lipase production by thermo-alkalophilic Bacillus sp. 8C was increased by 6.36 fold compared to unoptimized conditions. Key words: Thermophilic, alkaline, ribose, FeCl 3 , methionine, sesame oil INTRODUCTIONLipases (triglycerol hydrolases (EC 3.1.1.3) are the hydrolytic enzymes belonging to serine hydrolase family and catalyze conversion and synthesis of esters from glycerol and fatty acids. Lipolytic reactions are highly chemo-, regio and or enantioselective and operate at the interface of oil and water (Nguyen et al., 2010). High quality and low energy consumption of lipase catalyzed reactions make them superior over conventional steam splitting methods operating at high temperature and pressure (Treichel et al., 2010). These characteristics of lipases increased their demand in different industries including food and dairy, detergent, cosmetics, leather, paper and pulp, biodiesel and pharmaceutical (Hasan et al., 2007).Thermophilic lipases are of special interest for industrial applications due to their stability and high activity at higher temperatures and being stable in presence of chemicals (Messias et al., 2009;Uttatree et al., 2010). Lipases being third largest selling group of enzymes with more than 1000 t of annual production and approaching to a billion dollar business. Lipases active at alkaline pH are of great importance in detergent industry where these enzymes are used for removal of fat stains and dirt (Bora and Bora, 2012). The use of alkaline lipases in detergent formulation substantially

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“…Thermophilic Bacillus sp. have previously shown the lipase production at 50-60ºC (Kim et al, 2000;Bora and Bora, 2012). However, the reports on lipase production by Bacillus sp.…”

Section: Resultsmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Optimization of Lipase Production by Thermo-Alkalophilic Bacillus sp. 8C

Bhosale¹,

Uzma²,

Bismile³

2015

Research J. of Microbiology

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“…The same results were observed by Salihu et al (2011), who reported a significant reduction in the lipolytic activity of Candida cylindracea after the insertion of NH 4 Cl to the culture medium. In the study by Bora and Bora (2012), it was reported that various inorganic nitrogen sources including ammonium sulphate and urea enhanced lipase production of Bacillus sp. LBN2 in the range of 10-17 U mLG 1 .…”

Section: Resultsmentioning

confidence: 99%

Partial Purification and Characterization of a Thermoalkalophilic Lipase Originated from Bacillus atrophaeus FSHM2 and its Application for Ester Synthesis

Ameri¹,

Shakibaie²,

Amirpour-R³

et al. 2015

Biotechnology

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A B S T R A C T A thermoalkalophilic lipase producing bacterial strain, identified asBacillus atrophaeus FSHM2 using 16S rDNA sequencing analysis was isolated from salty soil and its lipase was partially purified and characterized. The obtained results revealed that glucose, hazelnut oil, urea and calcium ion positively affected the lipase production by increasing the lipolytic activity to 13582.5, 6270, 4442 and 5505 U LG 1 , respectively compared to that of basal medium (4150 U LG 1 ). The partially purified lipase acted optimally at pH 9 and retained 88.2% of its initial activity after 1 h of incubation at 100°C. A two fold increase in the relative activity of the partially purified lipase was obtained in the presence of 4 M of NaCl. Application of the partially purified lipase for the synthesis of ethyl and methyl valerate in the organic solvent medium (xylene) resulted in 81.6 and 62.4% esterification, respectively, after 24 h of incubation.

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“…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.…”

Section: Introductionmentioning

confidence: 99%

“…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).…”

Section: Introductionmentioning

confidence: 99%

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

Niyonzima

2014

Biologia

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The possibility of using Bacillus flexus XJU-1 lipase in detergent preparations was studied. The enzyme was monomeric protein as confirmed by liquid chromatography-mass spectrometry and its molecular weight was 15.95 kDa. The lipase showed optimum activity at pH 10.0 and was 100% stable for 24 h at pH 10.0 and 11.0. It exhibited maximum activity at 70• C and retained more than 70% of the initial activity at 60, 70 and 80• C for 24 h. The activity was stimulated by Ca 2+ , Ba 2+ , Mg 2+ and Co 2+ , whereas 50% of the initial activity was lost with Fe 3+ and Hg 2+ . The activity was inhibited by 10 mM N-bromosuccinimide and tosyl-L-lysylchloromethylketone, while N-ethylmaleimide, phenylmethylsulphonylfluoride and urea did not show any effect. The enzyme significantly hydrolysed olive, cottonseed, sunflower, groundnut, and gingelly oils. With p-nitrophenyl palmitate, Vmax and Km were 62.5 U/mL and 2.25 mM, respectively. The lipase maintained its stability in Tween-80, Triton-100 and H2O2 at 1%, but an activation of 10% and a reduction of 15% in relative activity were observed with NaClO and sodium dodecyl sulphate, respectively. The enzyme retained maximum storage stability for 20 days at −20, 4 and 30• C. In the presence of 0.7% (w/v) Ariel, Henko, Super wheel, Tide plus and Rin, a retention of more than 84.90% initial activity was recorded after 24 h at 60• C. The supplementation of the lipase to the detergents improved the olive oil stain removal. These properties suggested the present enzyme as a potential additive for detergent preparations.

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Optimization of extracellular thermophilic highly alkaline lipase from thermophilic Bacillus sp isolated from Hotspring of Arunachal Pradesh, India

Cited by 76 publications

References 35 publications

Optimization of Lipase Production by Thermo-Alkalophilic Bacillus sp. 8C

Optimization of Lipase Production by Thermo-Alkalophilic Bacillus sp. 8C

Partial Purification and Characterization of a Thermoalkalophilic Lipase Originated from Bacillus atrophaeus FSHM2 and its Application for Ester Synthesis

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

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