Immobilization of lipase enzyme extracted from thermophilic Bacillus licheniformis 14T local isolate

Saad Hussein Khudhair, Melad Khalaf Mohammed, Ahmed Darweesh Jabbar

Abstract


Background: Currently, lipase enzymes are considered important bio-catalysts in many industries due to their unique properties in catalyzing various types of reactions in aqueous solutions.  By targeting hydrocarbons in the oil, lipase enzymes contribute to the breakdown of hydrocarbons, reducing the environmental impact of oil spills and facilitating the remediation of contaminated areas.

Methods: A thermostable lipase from local isolate Bacillus licheniformis 14T has been immobilized on four different supports that include the inactivated chitosan beads, activated chitosan beads with glutaraldehyde, inactivated chitosan-alginate beads, and activated chitosan-alginate beads with glutaraldehyde.

Results The purified free lipase enzyme exhibited the highest enzymatic activity at 34.6 units/ml, surpassing all immobilized enzymes. Specific activity increased to 96.25 and 79.03 unit/mg protein for activated chitosan beads and activated chitosan-alginate beads, while decreasing to 52.95 and 46.05 unit/mg protein for inactivated chitosan beads and inactivated chitosan-alginate beads compared to the free enzyme. Optimal conditions for the immobilized enzyme differed, with the highest enzyme activity achieved after 60 minutes at 60°C and pH 8, reaching 48.6, 70.23, 43.12, and 61.2 units/ml on various supports, contrasting with the free enzyme's peak activity after 30 minutes at 50°C and pH 7.

Conclusions: Immobilizing the lipase enzyme increases the specific activity of the immobilized enzyme on the supports of activated chitosan beads with, also the immobilization process led to a change in the optimal conditions for the activity of the immobilized enzyme compared with the optimal conditions of free enzyme.

Keywords: Lipase; Bacillus licheniformis; Immobilization; Chitosan; Alginate   


Full Text:

PDF

References


Patel K, Parikh S. Identification, production, and purification of a novel lipase from Bacillus safensis. Journal of Applied Biology and Biotechnology, (2022); 10(4): 73-76.

Nawani N, Singh R, Kaur J. Immobilization and stability studies of a lipase from thermophilic Bacillus sp: The effect of process parameters on immobilization of enzyme. Electronic Journal of Biotechnology, (2006); 9(5): 559-565.

Kılınç A, Teke M, Önal S, Telefoncu A. Immobilization of pancreatic lipase on chitin and chitosan. Preparative biochemistry & biotechnology, (2006); 36(2): 153-163.

Bora L, Bora M. Optimization of extracellular thermophilic highly alkaline lipase from thermophilic Bacillus sp isolated from hotspring of Arunachal Pradesh, India. Brazilian Journal of Microbiology, (2012); 4330-42.

Ugras S. Characterization of a thermophilic lipase from Bacillus licheniformis Ht7 isolated from Hayran Thermal Springs in Giresun. Romanian Biotechnological Letters, (2017); 22(1): 12297-12306.

Chandra P, Enespa, Singh R, Arora PK. Microbial lipases and their industrial applications: a comprehensive review. Microbial cell factories, (2020); 19(169):1-42.

Rashid FAA, Rahim RA, Ibrahim D, Balan A, Bakar NMA. Purification and properties of thermostable lipase from a thermophilic bacterium, Bacillus licheniformis IBRL-CHS2. J Pure Appl Microbiol, (2013); 71635-1645.

Gohel H, Ghosh S, Bragazna V. Production, Purification and Immobilization of extracellular lipases from thermophilic Bacillus Subtilis XRF 11 and Bacillus Licheniformis XRF 12 FOR Production of alkyl esters. Int J Life Sc Bt & Pharm Res, (2013); 2 (3):278-286..

Sharma P, Sharma N, Pathania S, Handa S. Purification and characterization of lipase by Bacillus methylotrophicus PS3 under submerged fermentation and its application in detergent industry. Journal of Genetic Engineering and Biotechnology, (2017); 15(2): 369-377.

Carneiro LABC, Costa-Silva TA, Souza CRF, Bachmann L, Oliveira WP, Said S. Immobilization of lipases produced by the endophytic fungus Cercospora kikuchii on chitosan microparticles. Brazilian Archives of Biology and Technology, (2014); 57578-586.

Barik A, Sen SK, Rajhans G, Raut S. Purification and optimization of extracellular lipase from a novel strain Kocuria flava Y4. International Journal of Analytical Chemistry, (2022); 2022:1-10.

Egwim EC, Adeshina AA, Oyewole OA, Okoliegbe IN. Optimization of Lipase immobilised on Chitosan Beads for Biodiesel Production. (2012) ; 2(2):103-112..

Khudhair SH. Isolation and optimization of thermophilic lipase producing bacteria from soil contaminated with used engines oil. Journal of Wasit for Science and Medicine, (2016); 8(4): 66-74.

KHUDHAIR SH, MOHAMMED MK. PURIFICATION AND CHARACTERIZATION OF LIPASE ENZYME EXTRACTED FROM Bacillus licheniformis 14T LOCAL ISOLATE. PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY, (2021); 22(69-70): 169-177.

Sirisha E, Rajasekar N, Narasu ML. Isolation and optimization of lipase producing bacteria from oil contaminated soils. Advances in Biological Research, (2010); 4(5): 249-252.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. Journal of biological Chemistry, (1951); 193(1): 265-275.

Mendes AA, de Castro HF, de S. Rodrigues D, Adriano WS, Tardioli PW, et al. Multipoint covalent immobilization of lipase on chitosan hybrid hydrogels: influence of the polyelectrolyte complex type and chemical modification on the catalytic properties of the biocatalysts. Journal of Industrial Microbiology and Biotechnology, (2011); 38(8): 1055-1066.

Rodrigues DS, Mendes AA, Adriano WS, Gonçalves LR, Giordano RL. Multipoint covalent immobilization of microbial lipase on chitosan and agarose activated by different methods. Journal of Molecular Catalysis B: Enzymatic, (2008); 51(3-4): 100-109.

Mohamad NR, Marzuki NHC, Buang NA, Huyop F, Wahab RA. An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes. Biotechnology & Biotechnological Equipment, (2015); 29(2): 205-220.

Nazari T, Alijanianzadeh M, Molaeirad A, Khayati M. Immobilization of Subtilisin Carlsberg on modified silica gel by cross-linking and covalent binding methods. Biomacromolecular Journal, (2016); 2(1): 53-58.

Sharma Chander K, Kanwar Shamsher S. Purification of a Novel Thermophilic Lipase from B. licheniformis MTCC-10498. ISCA Journal of Biological Sciences, (2001);1, 43-48.

Saun NK, Mehta P, Gupta R. Purification and physicochemical properties of lipase from thermophilic Bacillus aerius. Journal of oleo science, (2014); 63(12): 1261-1268.

Sari ANM, Koentjoro MP, Prasetyo EN. Lipase Immobilization Based On Biopolymer. Surabaya international jealth conference; (2019); 406-418.

Carneiro EA, Bastos AKP, Oliveira UMFd, Matos LJBLd, Adriano WS, et al. Improving the catalytic features of the lipase from Rhizomucor miehei immobilized on chitosan-based hybrid matrices by altering the chemical activation conditions. Química Nova, (2020); 431234-1239.




DOI: http://dx.doi.org/10.62940/als.v11i2.2251

Refbacks

  • There are currently no refbacks.