Research Article | Volume 11, Issue 1, January, 2023

Bioesterification of carboxylic acids by immobilized esterase of Pisum sativum

Rajesh Dattatraya Tak Amol Ashok Bhosale Dnyaneshwar Dashrath Gaikwad   

Open Access   

Published:  Nov 22, 2022

DOI: 10.7324/JABB.2023.110106

Semi-purified immobilized esterase, isolated from the seed of Pisum sativum, was found to be an efficient biocatalyst for the facile esterification of pharmaceutically important carboxylic acids. The partially purified esterase precipitated using ammonium sulfate was immobilized using sodium alginate method. After incubation, the immobilized beads so formed were found to be spherical with an average size 3 mm. Immobilized esterase (Km: 580 ug, Vmax: 350 ug/min) to be tested for the esterification of aromatic carboxylic acids was incubated separately with methanol containing benzoic acid, 4-amino-2-chlorobenzoic acid, salicylic acid, and ethanol containing 4-aminobenzoic acid, respectively, at room temperature with constant stirring. Activity of free and immobilized esterase was measured with synthetic substrate, 4-nitrophenyl acetate. The reaction progress was monitored by thin layer chromatography (Hexane: Ethyl acetate) with successive time intervals. The corresponding esters methylbenzoate, 4-amino-2- chloromethylbenzoate, methylsalicylate, and 4-aminoethylbenzoate were analyzed with spectroscopic techniques and determination of physical constants. This confirmed the ability of esterase to transform organic acids smoothly into desired esters. The present studies demonstrated the suitability of preparation and promising procedure for the green synthesis of aromatic esters. Highly purified esterase of P. sativum will afford wide futuristic scope for large scale production of pharmaceutically and/or industrially important products.

Keyword:     Biotransformation Esterification Immobilized esterase Pisum sativum


Tak RD, Bhosale AV, Gaikwad DD. Bioesterification of carboxylic acids by immobilized esterase of Pisum sativum. J App Biol Biotech. 2023;11(1):51-54.

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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1. Frankel TN, Garber ED. Esterases in extracts from germinating seeds of twelve pea varieties. Int J Plant Sci 1965;126:221-2.

2. Bornscheuer UT. Microbial carboxyl esterases: Classification, properties and application in biocatalysis. FEMS Microbiol Rev 2002;26:73-81.

3. a) Bornscheuer UT, Kazlauskas RJ. Hydrolyses in organic synthesis: Regio-and stereoselective biotransformations. Chembiochem 2006;7:1279-80. b) Faber K. Biotransformations in Organic Chemistry. 4th ed. Berlin, Heidelberg: Springer-Verlag; 2011.

4. a) Giuliani S, Piana C, Setti L, Hochkoeppler A, Pifferi PG, Williamson G, et al. Synthesis of pentylferulate by a feruloyl esterase from Aspergillus niger using water-in-oil microemulsions. Biotechnol Lett 2001;23:325-30. b) Matsuo T, Kobayashi T, Kimura Y, Tsuchiyama M, Oh T, Sakamoto T, et al. Synthesis of glycerylferulate by immobilized ferulic acid esterase. Biotechnol Lett 2008;30:2151-6. c) Thörn C, Gustafsson H, Olsson L. Immobilization of feruloyl esterases in mesoporous materials leads to improved transesterification yield. J Mol Catal B Enzym 2011;72:57-64.

5. a) Antonopoulou I, Varriale S, Topakas E, Rova U, Christakopoulos P, Faraco V. Enzymatic synthesis of bioactive compounds with high potential for cosmeceutical application. Appl Microbiol Biotechnol 2016;100:6519-43. b) Faulds CB. What can Feruloylesterases do for us? Phytochem Rev 2010;9:121-32.

6. Kaushik N, SoumitraBiswas S, Singh J. Biocatalysis and biotransformation processes-an insight. Sci Tech Rev 2014;1:15.

7. Zhang DH, Yuwen LX, Peng LJ. Parameters affecting the performance of immobilized enzyme. J Chem 2013;2013:1-7. 8. Simpson DM, Beynon RJ. Acetone precipitation of proteins and the modification of peptides. J Proteome Res 2010;9:444-50.

9. Bradford MM. A rapid and sensitive for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54.

10. a) Panesar R, Panesar PS, Singh RS, Bera MB. Applicability of alginate entrapped yeast cells for the production of lactose-hydrolyzed milk. J Food Proc Eng 2007;30:472-84. b) Klibanov AM. Immobilized enzymes and cells as practical catalysts. Science 1983;219:722-7.

11. a) Pauly J, Gröger H, Patel AV. Design, characterisation and application of alginate-based encapsulated pig liver esterase. J Biotechnol 2018;280:42-8. b) Zakeri M, Moghadam H, Samimi A, Mohebbi-kalhori D. Optimization of calcium alginate beads production by electrospray using response surface methodology. Mater Res Express 2019;6:1-17.

12. Verma ML, Naebe M, Barrow CJ, Puri M. Enzyme immobilisation on amino-functionalised multi-walled carbon nanotubes: Structural and biocatalytic characterisation. PLoS One 2013;8:e73642.

13. Sörensen MH, Ng JB, Bergström L, Alberius PC. A improved enzymatic activity of Thermomyces lanuginosus lipase immobilized in a hydrophobic particulate mesoporous carrier. J Colloid Interface Sci 2010;343:359-65.

14. Ansari SA, Husain Q. Potential applications of enzymes immobilized on/in nanomaterials: A review. Biotechnol Adv 2012;30:512-23.

15. Mahajan R, Gupta VK, Sharma J. Comparison and suitability of gel matrix for entrapping higher content of enzymes for commercial applications. Indian J Pharm Sci 2010;72:223-8.

16. Vaija J, Linko YY, Linko P. Citric acid production with alginate bead entrapped Aspergillus niger ATCC 9142. Appl Biochem Biotechnol 1982;7:51-4.

17. Faccio G. From protein features to sensing surfaces. Sensor 2018;18:1204.

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