Study of Catalytic Activity of Lipase and Lipase-Chitosan Complexes in Dynamics

Authors

  • Anastasia A. Savina Chemistry Department, Federal State Budgetary Educational Institution of Higher Education “Moscow State Academy of Veterinary Medicine and Biotechnology - MVA after K.I. Skryabin”, 109472 Moscow, Russian Federation
  • Olga V. Abramova Chemistry Department, Federal State Budgetary Educational Institution of Higher Education “Moscow State Academy of Veterinary Medicine and Biotechnology - MVA after K.I. Skryabin”, 109472 Moscow, Russian Federation
  • Lilia S. Garnashevich Chemistry Department, Federal State Budgetary Educational Institution of Higher Education “Moscow State Academy of Veterinary Medicine and Biotechnology - MVA after K.I. Skryabin”, 109472 Moscow, Russian Federation
  • Ilia S. Zaitsev Chemistry Department, Federal State Budgetary Educational Institution of Higher Education “Moscow State Academy of Veterinary Medicine and Biotechnology - MVA after K.I. Skryabin”, 109472 Moscow, Russian Federation
  • Oksana A. Voronina Chemistry Department, Federal State Budgetary Educational Institution of Higher Education “Moscow State Academy of Veterinary Medicine and Biotechnology - MVA after K.I. Skryabin”, 109472 Moscow, Russian Federation
  • Marina S. Tsarkova Chemistry Department, Federal State Budgetary Educational Institution of Higher Education “Moscow State Academy of Veterinary Medicine and Biotechnology - MVA after K.I. Skryabin”, 109472 Moscow, Russian Federation
  • Sergei Yu. Zaitsev Chemistry Department, Federal State Budgetary Educational Institution of Higher Education “Moscow State Academy of Veterinary Medicine and Biotechnology - MVA after K.I. Skryabin”, 109472 Moscow, Russian Federation

DOI:

https://doi.org/10.6000/1929-5995.2019.08.03

Keywords:

Lipase, chitosan, enzyme immobilization, activity, triacetin

Abstract

Pancreatic porcine lipase (PPL) is the unique enzyme in numerous biochemical processes for human and animals. Stability in time is the critical point for many enzymes in relation to their further applications. The effect of chitosan on the activity of PPL during 2 months was investigated.

Materials and Methods: Potentiometric method was used to study the catalytic activity of enzymes that based on measuring of the potential an electrode immersed in a triacetin (substrate) solution by titration with 0.01 M NaOH. A laboratory pH-stat with combined pH electrode was used for the measurements of this potential in the presence of PPL or PPL-chitosan complexes. Standard experimental conditions: 40 ºC, pH 7.0.

Results: The following PPL activity values (data given in % to the activity of free lipase) vs. time (in 10 min. intervals) were obtained: 100%, 97.4%, 89.9%, 82.9%, 77.9% and 75.9% after 10, 20, 30, 40, 50 and 60 min., respectively. The PPL catalytic activity decreased at about ¼ to ½ of the initial values after 7 or 14 days, 1 or 2 months storage, although there were differences in the decline dynamics. The complex PPL:Chit=50:1 has better stabilizing properties as compared to other complexes; does not strongly inhibit lipase and requires a small amount of carrier (chitosan) for its formation.

Conclusions: The complex PPL:Chit=25:1 is less effective as the PPL:Chit=50:1 complex, but both can be applicable in some industrial processes.

References

Sharma R, Chisti Y, Banerjee UC. Production, purification, characterization, and applications of lipases Biotechnology Advances 2001; 19: 627-662. https://doi.org/10.1016/S0734-9750(01)00086-6 DOI: https://doi.org/10.1016/S0734-9750(01)00086-6

Vulfson EN. Industrial application of lipases. In: Woolley P. Peterson SB. Editors. Lipases – their structure, biochemistry and applications. Cambridge Univ. Press; 1994; 271-288.

Jaeger K-E, Reetz MT. Microbial lipases form versatile tools for biotechnology. Trends in Biotechnology 1998; 16(9): 396-403. https://doi.org/10.1016/S0167-7799(98)01195-0 DOI: https://doi.org/10.1016/S0167-7799(98)01195-0

Jaeger K-E, Dijkstra BW, Reetz MT. Bacterial biocatalysts: Molecular Biology, Three-Dimensional Structures, and Biotechnological Applications of Lipases. Annual Review of Microbiology 1999; 53(1): 315-351. https://doi.org/10.1146/annurev.micro.53.1.315 DOI: https://doi.org/10.1146/annurev.micro.53.1.315

Zaitsev SYu. Supramolecular Nanosized Systems at the Phase Interface: Concepts and Prospects for Bio-Nanotechnologies. Moscow: LENAND; 2010. [in Russian]

Eremeev NL, Zaitsev SY. Рorcine pancreatic lipase as a catalyst in organic synthesis. Mini-Reviews in Organic Chemistry 2016; 13(1): 78-85. https://doi.org/10.2174/1570193X13666160225000520 DOI: https://doi.org/10.2174/1570193X13666160225000520

Sizhu Ren, Conghai Li, Xiaobo Jiao, Shiru Jia, Jiandong Cui. Recent progress in multienzymes co-immobilization and multienzyme system applications. Chemical Engineering Journal 2019; 373(1): 1254-1278. https://doi.org/10.1016/j.cej.2019.05.141 DOI: https://doi.org/10.1016/j.cej.2019.05.141

De Caro J, Boudouard M, Bonicel J, Guidoni A, Desnuelle P, Rovery M. Porcine pancreatic lipase. Completion of the primary structure. Biochimica et Biophysica Acta 1982; 671(2): 129-38. https://doi.org/10.1016/0005-2795(81)90126-4 DOI: https://doi.org/10.1016/0005-2795(81)90126-4

Zaitsev SYu. Biological chemistry: from biologically active substances to organs and tissues of animals. Moscow: Capital Print Publishing; 2017.

Chapus C, Rovery M, Sarda L, Verger R. Pancreatic Lipase and Colipase, Biochimie 1988; 70: 1223-1234. https://doi.org/10.1016/0300-9084(88)90188-5 DOI: https://doi.org/10.1016/0300-9084(88)90188-5

Reis P., Holmberg K., Watzke H., Leser M.E., Miller R. Lipases at interfaces: A review. Advances in Colloid and Interface Science 2009;147–148: 237–250. https://doi.org/10.1016/j.cis.2008.06.001 DOI: https://doi.org/10.1016/j.cis.2008.06.001

Ivanov EA, Aha B, Volchenkova TA, Zaitsev SYu, Investigation Of Enzymatic Hydrolysis of Lipid-Like Substrates and Trilaurin in Monolayers. Colloids and Surfaces B: Biointerfaces 2002; 23(4): 349-356. https://doi.org/10.1016/S0927-7765(01)00265-X DOI: https://doi.org/10.1016/S0927-7765(01)00265-X

Zaitsev SYu, Gorokhova IV, Cashtigo TV, Zintchenco A, Dautzenberg H, General Approach for Lipases Immobilization in Polyelectrolyte Complexes. Colloids and Surfaces A: Physicochemical and Engineering Aspects 2003; 221(1-3): 209-220. https://doi.org/10.1016/S0927-7757(03)00137-7 DOI: https://doi.org/10.1016/S0927-7757(03)00137-7

Verger R. “Interfacial activation” of lipases: facts and artifacts. Trends in Biotechnology 1997; 151: 32-38. https://doi.org/10.1016/S0167-7799(96)10064-0 DOI: https://doi.org/10.1016/S0167-7799(96)10064-0

Savina AA, Salahuddin A, Zaitsev SYu. Changes in the activity of lipase from porcine pancreas in combination with polystyrene microsphere of different diameters. Veterinariya, zootekhniya and biotekhnologiya 2015; 12: 63-67.

Savina AA, Solovyovа DO, Zaitsev SYu. Formation and Properties of The Polyelectrolyte Complexes of Polylysine with Porcine Pancreatic Lipase. Veterinariya, zootekhniya and biotekhnologiya 2018; 8: 46-50.

Cheung RCF, Ng TB, Wong JH, Chan WY. Chitosan: An Update on Potential Biomedical and Pharmaceutical Applications. Marine Drugs 2015; 13(8): 5156-5186. https://doi.org/10.3390/md13085156 DOI: https://doi.org/10.3390/md13085156

Younes I, Rinaudo M. Chitin and Chitosan Preparation from Marine Sources. Structure, Properties and Applications. Marine Drugs 2015; 13(3): 1133-1174. https://doi.org/10.3390/md13031133 DOI: https://doi.org/10.3390/md13031133

Pellá MCG, Lima-Tenório MK, Tenório-Neto ET, Guilherme MR, Muniz EC, Rubira AF. Chitosan-based Hydrogels: From Preparation to Biomedical Applications. Carbohydrate Polymers 2018; 196: 233. https://doi.org/10.1016/j.carbpol.2018.05.033 DOI: https://doi.org/10.1016/j.carbpol.2018.05.033

Ruocco N, Costantini S, Guariniello S, Costantini M. Polysaccharides from the Marine Environment with Pharmacological, Cosmeceutical and Nutraceutical Potential. Molecules 2016; 21: 551. https://doi.org/10.3390/molecules21050551 DOI: https://doi.org/10.3390/molecules21050551

Majeti NV, Kumar R. A review of Chitin and Chitosan Applications. Reactive & Functional Polymers 2000; 46: 1-26. https://doi.org/10.1016/S1381-5148(00)00038-9 DOI: https://doi.org/10.1016/S1381-5148(00)00038-9

de Mello MD, Corderiro D, Costa LT, Follmer C, Catalytic properties of lipases immobilized onto ultrasound-treated chitosan supports. Biotechnology and bioprocess engineering 2013; 18(6): 1090-1100. https://doi.org/10.1007/s12257-013-0285-5 DOI: https://doi.org/10.1007/s12257-013-0285-5

Desai PD, Dave AM, Devi S, Alcoholysis of salicornia oil using free and covalently bound lipase onto chitosan beads Food chemistry 2006; 95(2): 193-199. https://doi.org/10.1016/j.foodchem.2004.12.030 DOI: https://doi.org/10.1016/j.foodchem.2004.12.030

Kılınç A, Teke M, Önal S, Telefoncu A, Immobilization of pancreatic lipase on chitin and chitosan. Preparative biochemistry & biotechnology 2006; 36(2): 153-163. https://doi.org/10.1080/10826060500533976 DOI: https://doi.org/10.1080/10826060500533976

Ozyilmaz G, Yağız E. Isoamylacetate production by entrapped and covalently bound Candida rugosa and porcine pancreatic lipases. Food chemistry 2012; 4: 2326-2332. https://doi.org/10.1016/j.foodchem.2012.07.062 DOI: https://doi.org/10.1016/j.foodchem.2012.07.062

Pereira EB, Zanin GM, Castro HF, Immobilization and catalytic properties of lipase on chitosan for hydrolysis and esterification reactions. Brazilian Journal of Chemical Engineering 2003; 4: 343-355. https://doi.org/10.1590/S0104-66322003000400002 DOI: https://doi.org/10.1590/S0104-66322003000400002

Gilani SL, Najafpour GD, Moghadamnia A, Kamaruddin AH. Stability of Immobilized Porcine Pancreas Lipase on Mesoporous Chitosan Beads: A Comparative Study. Journal of Molecular Catalysis B: Enzymatic 2016; 133: 144-153. https://doi.org/10.1016/j.molcatb.2016.08.005 DOI: https://doi.org/10.1016/j.molcatb.2016.08.005

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Published

2019-08-27

How to Cite

Savina, A. A., Abramova, O. V., Garnashevich, L. S., Zaitsev, I. S., Voronina, O. A., Tsarkova, M. S., & Zaitsev, S. Y. (2019). Study of Catalytic Activity of Lipase and Lipase-Chitosan Complexes in Dynamics . Journal of Research Updates in Polymer Science, 8, 15–20. https://doi.org/10.6000/1929-5995.2019.08.03

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