Activity and Stability of Lipases Immobilized onto Acetylated Bacterial Cellulose
Article Sidebar
Main Article Content
Abstract
Bacterial cellulose (BC) materials were used for lipase immobilization to improve enzyme activity and stability. BC films produced by Komagataeibacter xylinus were first acetylated in an acetic anhydride/iodine system to convert their OH groups to more hydrophobic acetyl groups. Activity yield (44.4%) and maximum specific activity (12.44 μmol mg–1 min–1) were achieved when 400 mg of BC was acetylated in 20 mL of acetic anhydride containing 0.275 mM of iodine. Studies on the catalytic activity of lipase also show that the immobilization of lipase on acetylated BC (ABC) films significantly enhanced its tolerance to temperature and pH. Immobilized lipases retained 89% and 56% of their catalytic activities after being incubated at 60 °C and 80 °C for 1 h, respectively; while those of free lipases significantly decreased to 24% (60 °C) and only 11% (80 °C). Immobilized lipases incubated at pH 5.0 and pH 10.0 for 24 h also retained high catalytic activities (70% and 82%, respectively), considerably higher than those of free lipases (19% - pH 5.0 and 63% - pH 10.0). Tolerance to organic solvents, such as n-hexane, acetone, ethanol, isopropanol of ABC-immobilized lipase was also improved. The immobilization of lipase on ABC films significantly improved its reusability and storage stability: ABC-immobilized lipase still could be reused for 30 cycles with residual activities of more than 90%, and still retained 95% of its early activity after 15-day storage at 4 °C. This implies that ABC-immobilized lipase is potentially applied in food, medicine, biodiesel and detergent industries.
Article Details
References
E. L. Bell, W. Finnigan, S. P. France, A. P. Green, M. A. Hayes, L. J. Hepworth, S. L. Lovelock, H. Niikura, S. Osuna, E. Romero, K. S. Ryan, N. J. Turner, and S. L. Flitsch, “Biocatalysis,” Nature Reviews Methods Primers, vol. 1, no. 1, p. 46, 2021.
R. C. Rodrigues, J. J. Virgen-Ortíz, J. C. S. dos Santos, Á. Berenguer-Murcia, A. R. Alcantara, O. Barbosa, C. Ortiz, and R. Fernandez-Lafuente, “Immobilization of lipases on hydrophobic supports: Immobilization mechanism, advantages, problems, and solutions,” Biotechnology Advances, vol. 37, no. 5, pp. 746–770, 2019.
R. C. Alnoch, L. Alves dos Santos, J. Marques de Almeida, N. Krieger, and C. Mateo, “Recent trends in biomaterials for immobilization of lipases for application in non-conventional media,” Catalysts, vol. 10, no. 6, p. 697, 2020.
P. S. Mateos, M. B. Navas, S. R. Morcelle, C. Ruscitti, S. R. Matkovic, and L. E. Briand, “Insights in the biocatalyzed hydrolysis, esterification and transesterification of waste cooking oil with a vegetable lipase,” Catalysis Today, vol. 372, pp. 211–219, 2021.
P. Chandra, Enespa, R. Singh, and P. K. Arora, “Microbial lipases and their industrial applications: A comprehensive review,” Microbial Cell Factories, vol. 19, no. 1, p. 169, 2020.
J. J. Virgen-Ortíz, J. C. S. dos Santos, C. Ortiz, Á. Berenguer-Murcia, O. Barbosa, R. C. Rodrigues, and R. Fernandez-Lafuente, “Lecitase ultra: A phospholipase with great potential in biocatalysis,” Molecular Catalysis, vol. 473, Art. no. 110405, 2019.
L. Urbina, M. Á. Corcuera, N. Gabilondo, A. Eceiza, and A. Retegi, “A review of bacterial cellulose: Sustainable production from agricultural waste and applications in various fields,” Cellulose, vol. 28, no. 13, pp. 8229–8253, 2021.
H. M. C. Azeredo, H. Barud, C. S. Farinas, V. M. Vasconcellos, and A. M. Claro, “Bacterial cellulose as a raw material for food and food packaging applications,” Frontiers in Sustainable Food Systems, vol. 3, 2019, doi: 10.3389/ fsufs.2019.00007.
A. A. N. Oliveira, E. d. F. M. d. Mesquita, and A. A. L. Furtado, “Use of bacterial cellulose as a fat replacer in emulsified meat products: Review,” Food Science and Technology, vol. 42, Art. no. e42621, 2021.
P. Paximada, E. Tsouko, N. Kopsahelis, A. A. Koutinas, and I. Mandala, “Bacterial cellulose as stabilizer of o/w emulsions,” Food Hydrocolloids, vol. 53, pp. 225–232, 2016.
L. Chen, M. Zou, and F. F. Hong, “Evaluation of fungal laccase immobilized on natural nanostructured bacterial cellulose,” Frontiers in Microbiology, Original Research, vol. 6, 2015, doi: 10.3389/fmicb.2015.01245.
S.-C. Wu, Y.-K. Lia, and C. Ho, “Glucoamylase immobilization on bacterial cellulose using periodate oxidation method,” International Journal of Science and Engineering, vol. 3, pp. 1–4, 2013.
W. Wang H.-Y. Li, D.-W. Zhang, J. Jiang, Y.-R. Cui, S. Qiu, Y.-L. Zhou, and X.-X. Zhang, “Fabrication of bienzymatic glucose biosensor based on novel gold nanoparticles-bacteria cellulose nanofibers nanocomposite,” Electroanalysis, vol. 22, no. 21, pp. 2543–2550, 2010.
P. Bayazidi, H. Almasi, and A. K. Asl, “Immobilization of lysozyme on bacterial cellulose nanofibers: Characteristics, antimicrobial activity and morphological properties,” International Journal of Biological Macromolecules, vol. 107, pp. 2544–2551, 2018.
R. Singla, A. Guliani, A. Kumari, and S. K. Yadav, “Nanocellulose and nanocomposites,” in Nanoscale Materials in Targeted Drug Delivery, Theragnosis and Tissue Regeneration, S. K. Yadav, Ed. Singapore: Springer Singapore, pp. 103–125, 2016.
Q. Cai, C. Hu, N. Yang, Q. Wang, J. Wang, H. Pan, Y. Hu, and C. Ruan, “Enhanced activity and stability of industrial lipases immobilized onto spherelike bacterial cellulose,” International Journal of Biological Macromolecules, vol. 109, pp. 1174–1181, 2018.
W. Hu, S. Chen, Q. Xu, and H. Wang, “Solventfree acetylation of bacterial cellulose under moderate conditions,” Carbohydrate Polymers, vol. 83, no. 4, pp. 1575–1581, 2011.
M. Nogi, K. Abe, K. Handa, F. Nakatsubo, S. Ifuku, and H. Yano, “Property enhancement of optically transparent bionanofiber composites by acetylation,” Applied Physics Letters, vol. 89, no. 23, Art. no. 233123, 2006.
X.-J. Huang, P.-C. Chen, F. Huang, Y. Ou, M.-R. Chen, and Z.-K. Xu, “Immobilization of Candida rugosa lipase on electrospun cellulose nanofiber membrane,” Journal of Molecular Catalysis B: Enzymatic, vol. 70, no. 3–4, pp. 95–100, 2011.
C. M. Soares, H. F. De Castro, F. F. De Moraes, and G. M. Zanin, “Characterization and utilization of Candida rugosa lipase immobilized on controlled pore silica,” Applied Biochemistry and Biotechnology, vol. 79, pp. 745–757, 1999.
I. B.-B. Romdhane, Z. B. Romdhane, A. Gargouri, and H. Belghith, “Esterification activity and stability of Talaromyces thermophilus lipase immobilized onto chitosan,” Journal of Molecular Catalysis B: Enzymatic, vol. 68, no. 3–4, pp. 230– 239, 2011.
V. Lima, N. Krieger, D. Mitchell, and J. Fontana, “Activity and stability of a crude lipase from Penicillium aurantiogriseum in aqueous media and organic solvents,” Biochemical Engineering Journal, vol. 18, no. 1, pp. 65–71, 2004.
G. Bayramoğlu and M. Y. Arıca, “Preparation of poly (glycidylmethacrylate–methylmethacrylate) magnetic beads: Application in lipase immobilization,” Journal of Molecular Catalysis B: Enzymatic, vol. 55, no. 1–2, pp. 76–83, 2008.
A. Mustranta, P. Forssell, and K. Poutanen, “Applications of immobilized lipases to transesterification and esterification reactions in nonaqueous systems,” Enzyme and Microbial Technology, vol. 15, no. 2, pp. 133–139, 1993.
P. Pinsirodom and K. L. Parkin, “Lipase assays,” Current Protocols in Food Analytical Chemistry, vol. 00, no. 1, pp. C3.1.1–C3.1.13, 2001, doi: 10.1002/0471142913.fac0301s00.
J. H. Waterborg, “The Lowry method for protein quantitation,” in The Protein Protocols Handbook Totowa. NJ: Humana Press, 2009, pp. 7–10.
D.-Y. Kim, Y. Nishiyama, and S. Kuga, “Surface acetylation of bacterial cellulose,” Cellulose, vol. 9, no. 3–4, pp. 361–367, 2002.
M. Božič, V. Vivod, S. Kavčič, M. Leitgeb, and V. Kokol, “New findings about the lipase acetylation of nanofibrillated cellulose using acetic anhydride as acyl donor,” Carbohydrate Polymers, vol. 125, pp. 340–351, 2015.
H. S. Barud, A. M. de Araújo Júnior, D. B. Santos, R. M. de Assunção, C. S. Meireles, D. A. Cerqueira, G. R. Filho, C. A. Ribeiro, Y. Messaddeq, and S. J. Ribeiro, “Thermal behavior of cellulose acetate produced from homogeneous acetylation of bacterial cellulose,” Thermochimica Acta, vol. 471, no. 1–2, pp. 61–69, 2008.
M. Jonoobi, J. Harun, A. P. Mathew, M. Z. B. Hussein, and K. Oksman, “Preparation of cellulose nanofibers with hydrophobic surface characteristics,” Cellulose, vol. 17, no. 2, pp. 299–307, 2010.
O. Yemul and T. Imae, “Covalent-bonded immobilization of lipase on poly (phenylene sulfide) dendrimers and their hydrolysis ability,” Biomacromolecules, vol. 6, no. 5, pp. 2809–2814, 2005.
M. Karra-Châabouni, I. Bouaziz, S. Boufi, A. M. B. do Rego, and Y. Gargouri, “Physical immobilization of Rhizopus oryzae lipase onto cellulose substrate: Activity and stability studies,” Colloids and Surfaces B: Biointerfaces, vol. 66, no. 2, pp. 168–177, 2008.
N. Kharrat, Y. B. Ali, S. Marzouk, Y.-T. Gargouri, and M. Karra-Châabouni, “Immobilization of Rhizopus oryzae lipase on silica aerogels by adsorption: Comparison with the free enzyme,” Process Biochemistry, vol. 46, no. 5, pp. 1083– 1089, 2011.
Y. İ. Doğaç, İ. Deveci, B. Mercimek, and M. Teke, “A comparative study for lipase immobilization onto alginate based composite electrospun nanofibers with effective and enhanced stability,” International Journal of Biological Macromolecules, vol. 96, pp. 302–311, 2017.
K. Vivek, G. S. Sandhia, and S. Subramaniyan, “Extremophilic lipases for industrial applications: A general review,” Biotechnology Advances, vol. 60, 2022, Art. no. 108002.
B. Hamid and F. A. Mohiddin, “Cold-active enzymes in food processing,” in Enzymes in Food Technology: Improvements and Innovations, M. Kuddus, Ed. Singapore: Springer, 2018, pp. 383–400.
M. Kavitha, “Cold active lipases – An update,” Frontiers in Life Science, vol. 9, no. 3, pp. 226–238, 2016.
H. A. Hasibuan, A. B. Sitanggang, N. Andarwulan, and P. Hariyadi, “Enzymatic synthesis of human milk fat substitute - A review on technological approaches,” Food Technol Biotechnol, vol. 59, no. 4, pp. 475–495, 2021.
Z. Zhang, S. Zhang, W. J. Lee, O. M. Lai, C. P. Tan, and Y. Wang, “Production of structured triacylglycerol via enzymatic interesterification of medium-chain triacylglycerol and soybean oil using a pilot-scale solvent-free packed bed reactor,” Journal of the American Oil Chemists' Society, vol. 97, no. 3, pp. 271–280, 2020.
A. G. A. SÁ, A. C. d. Meneses, P. H. H. d. Araújo, and D. d. Oliveira, “A review on enzymatic synthesis of aromatic esters used as flavor ingredients for food, cosmetics and pharmaceuticals industries,” Trends in Food Science and Technology, vol. 69, pp. 95–105, 2017.
E. H. Ahmed, T. Raghavendra, and D. Madamwar, “A thermostable alkaline lipase from a local isolate Bacillus subtilis EH 37: characterization, partial purification, and application in organic synthesis,” Applied Biochemistry and Biotechnology, vol. 160, no. 7, pp. 2102–2113, 2010.
X.-J. Huang, D. Ge, and Z.-K. Xu, “Preparation and characterization of stable chitosan nanofibrous membrane for lipase immobilization,” European Polymer Journal, vol. 43, no. 9, pp. 3710–3718, 2007.
S.-C. Wu, S.-M. Wu, and F.-M. Su, “Novel process for immobilizing an enzyme on a bacterial cellulose membrane through repeated absorption,” Journal of Chemical Technology and Biotechnology, vol. 92, no. 1, pp. 109–114, 2017.