Purification and Characterization of a Halotolerant And Thermotolerant Lipase Produced From a Novel Bacteria “Brevibacterium Halotolerans PS4 |KX671556|” and Its Application In Detergent Formulations
Keywords:
Extracellular lipase, Purification, Characterization, Brevibacterium halotolerans PS4
Abstract
An extracellular lipase was isolated and purified from the culture broth of Brevibacterium halotolerans PS4 to provide homogeneity using ammonium sulfate precipitation followed by chromatographic techniques on Sephadex G-75 colummn, resulting in a purification factor of 2.98 fold with specific activity of 1016.10 IU/mg. The molecular weight of the purified lipase was estimated by SDS-PAGE to be 80 kDa. The purified lipase had the maximal activity within the pH range of 6 to 7, with an optimum pH of 7, and within the temperature range of 35 to 55°C. The purified lipase exhibited not only stable but enhanced maximal activity by Triton X100. The enzyme activity of Brevibacterium halotolerans PS4 lipase was enhanced by Ca2+ and Mg2+. SDS and metal ions such as Hg2+, Zn2+, Cu2+, Ag2+ and Fe2+ decreased the lipase activity remarkedly. The extracellular lipase from orchard soil isolate was further applied for its application as laundary additives.References
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Bae J H, Kwon M H, Kim I H, Hou C T and Kim H R (2014) Purification and characterization of a cold-active lipase from Pichia lynferdii Y-7723: pH-dependant activity deviation Biotechnol Bioprocess Eng 19 851-857
Bano S, Syed M.N, Iqbal S, Khan A B, Ali-ul-Qader S and Azhar A (2010) Purification and characterization of 60 kD lipase linked with chaperonin from Pseudomonas aeruginosa BN-1 Afr J Biotechnol 9 724-7732
Bora L and Kalita MC (2008). Production of thermostable alkaline lipase on vegetable oils from a thermophilic Bacillus sp. DH4, characterization and its potential applications as detergent additive J Chem Technol Biot 83 688-693
Chigusa K, Hasegawa T, Yamamoto N and Watanabe Y (1996). Treatment of wastewater from oil manufacturing plant by yeasts Water Sci Technol 34 51-58
Daoud F B O, Kaddour S and Sadoun T (2013) Adsorption of cellulase Aspergillus niger on a commercial activated carbon: kinetics and equilibrium studies Colloid Surf B Biointerfaces 75 93–99
Garlapati V K, Vundavilli P R and Banerjee R (2010) Evaluation of lipase production by genetic algorithm and particle swarm optimization and their comparative study Appl Biochem Biotechnol 162 1350-1361
Ghasemi S, Rasoul A, Kazemi G, Zarrini, Morowvat M H and Kargar M (2015) Isolation and characterization of some moderately halophilic bacteria with lipase activity Microbiol 80 483-487
Jaeger K E and Reetz T M (1998) Microbial lipases from versatile tools for biotechnology Trends Biotechnol 16 396–403
Kiran K K and Chandra T S (2008) Production of surfactant and detergent-stable, halophilic, and alkalitolerant alpha-amylase by a moderately halophilic Bacillus sp. strain TSCVKK Appl Microbiol Biotechnol 77 1023-1031
Kumar S G, Gurramkonda C, Rather G, Muniramanna G S C, Mangamuri U K, Podha S and Choi Y L (2016) Glucoamylase from a newly isolated Aspergillus niger FME: detergent-mediated production, purification and characterization. J Korean Soc Appl Biol Chem 56 427−433
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Lowry O H, Rosebrough N J, Farr A L and Randall R J (1951) Protein measurement with the Folin-Phenol Reagent J Biol Chem 193 265-275
Mazhar H, Abbas N, Ali S S, Hussain Z and Ali S (2016) Purification and Characterization of Lipase Production from Bacillus subtilis PCSIR-NL39 J Biol Chem Research 33 546-558
Nadia N, Nehad Z A, Elsayed A E, Essam M A and Hanan M A (2010) Optimization of lipase synthesis by Mucor racemosus - Production in a triple impeller bioreactor Malaysian J Microbiol 6 7-15
Nawani N, Dosanjh N S and Kaur J (1998) A novel thermostable lipase from a thermophilic Bacillus sp.: characterization and esterification studies Biotechnol Lett 20 997–1000
Novak J, Kralova B, Demnerova K, Prochazka K, Vodrazka Z, Tolman J, Rysova D, Smidrkal J and Lopata V (1990) Enzyme agent based on lipasesand oxidoreductases for washing, degreasing and water reconditioning European Patent 355 228
Padmapriya B, Rajeswari T, Noushida E, Sethupalan D G and Venil C K (2011) Production of Lipase Enzyme from Lactobacillus spp. and Its Application in the Degradation of Meat World Appl Sci J 12 1798-1802
Rajesh E M, Arthe R, Rajendran R, Balakumar C, Pradeepa N and Anitha S (2010) Investigation of lipase production by Trichoderma reesei and optimization of production parameters J Environ Agric Food Chem 9 1177-1189
Ramani K, Kennedy LJ, Ramakrishnan M and Sekaran G (2010) Purification, characterization and application of acidic lipase from Pseudomonas gessardii using beef tallow as a substrate for fats and oil hydrolysis Process Biochem 45 1683-1691
Sebdani R M, Ardakani M R, Ghezelbash G R and Sadrinasab M (2011) Phylogenic Characterization of Lipase Producing Bacillus Strains Isolated from Persian Gulf Sediments Aust J Basic Appl Sci 5 121-126
Singh R, Tripathi R D, Dwivedi S, Kumar A, Trivedi P K and Chakrabarty D (2014) Lead bioaccumulation potential of an aquatic macrophyte Najas indica are related to antioxidant system Bioresour Technol 101 3025–3032
Sirisha E, Rajasekar N and Lakshmi M N (2010) Isolation and optimization of lipase producing bacteria from oil contaminated soils Advan Biol Res 4 249-252
Tan T, Zhang M, Wang B, Ying C and Deng I (2003) Secreening of high lipase producing Candida sp and production of lipase by fermentation Process Biochem 39 459-465
Tripathi P, Niyonzima F N and More S (2013) Detergent-compatible proteases: microbial production, properties, and stain removal analysis Prep Biochem Biotechnol 45 233-258
