Research Article | Volume 10, Supplement 2, July, 2022

Extraction and quantification of acrylic acid from acrylamidase-catalyzed reaction produced by Bacillus tequilensis

Riddhi Prabha Vinod Kumar Nigam   

Open Access   

Published:  Jun 20, 2022

DOI: 10.7324/JABB.2022.10s208

The reduction of non-renewable material has raised significant concerns for several years over the sustainable production of organic acids through bio-based methods in the world. One such way to overcome this problem is to use reactive extractants, in which appropriate extractants are employed to recover various organic and inorganic acids. The extraction of acrylic acid by solvent extraction is an illustration of this technique. The current study focuses on the synthesis of acrylic acid from acrylamidase produced by Bacillus tequilensis, succeeded by acid extraction from the amidase-catalyzed reaction by a solvent technique. Among the various solvents, ethyl acetate (2:1, v/v) was established as the most appropriate solvent for the extraction. Exactly 65 mg of raw acrylic acid was recovered from 20 ml of the amidase-catalyzed reaction. Various analytical methods such as thin layer chromatography, fourier transform infrared spectroscopy, high-performance liquid chromatography (HPLC), and mass spectrophotometry were accomplished for the identification, validation, and quantification of the extracted acrylic acid. The m/z value of acrylic acid obtained in the extracted product was 73.18, which was similar to the standard acrylic acid. From HPLC, almost 34% of bioconversion was quantified (3.4 mM) from 10 mM of acrylamide consumed. The extracted acrylic acid can be further exploited as chemical intermediates and pharmaceuticals in the future.

Keyword:     Acrylic acid acrylamide Bacillus tequilensis reactive extractant acrylamidase biocatalysis


Prabha R, Nigam VK. Extraction and quantification of acrylic acid from acrylamidase-catalyzed reaction produced by Bacillus tequilensis. J Appl Biol Biotech 2022;10(Suppl 2):78-84.

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. Reyhanitash E, Zaalberg B, Ijmker HM, Kersten SR, Schuur B. CO2- enhanced extraction of acetic acid from fermented wastewater. Green Chem 2015;17(8):4393-400; doi: 10.1039/C5GC01061F

2. Zhou Z, Li Z, Qin W. Reactive extraction of saturated aliphatic dicarboxylic acids with trioctylamine in 1-octanol: equilibria, model, and correlation of apparent reactive equilibrium constants. Ind Eng Chem Res 2013;52(31):10795-801; doi: 10.1021/ie4008002

3. Straathof AJ, Sie S, Franco TT, Van der Wielen LA. Feasibility of acrylic acid production by fermentation. Appl Microbiol Biotechnol 2005;67(6):727-34; doi: 10.1007/s00253-005-1942-1

4. Xiaobo XU, Jianping L, Peilin C. Advances in the research and development of acrylic acid production from biomass. Chinese J Chem Eng 2006;14(4):419-27; doi: 10.1016/S1004-9541(06)60094-3

5. Kumar S, Pandey S, Wasewar KL, Ak N, Uslu H. Reactive extraction as an intensifying approach for the recovery of organic acids from aqueous solution: a comprehensive review on experimental and theoretical studies. J Chem Eng Data 2021;66(4):1557-73; doi: 10.1021/acs.jced.0c00405

6. Altunay N, Elik A, Gürkan R. Extraction and reliable determination of acrylamide from thermally processed foods using ionic liquid-based ultrasound-assisted selective microextraction combined with spectrophotometry. Food Addit Contam Part A 2018;35(2):222-32; doi: 10.1080/19440049.2017.1394585

7. Chen J, Zheng RC, Zheng YG, Shen YC. Microbial transformation of nitriles to high-value acids or amides. Biotechnol China I 2009; 113: 33-77. doi: 10.1007/10_2008_25

8. Tuyun AF, Uslu H. Reactive extraction of acrylic acid using trioctylamine (TOA) in versatile diluents. Desalin Water Treat 2015;55(1):193-8; doi: 10.1080/19443994.2014.911707

9. Kar A, Bagde A, Athankar KK, Wasewar KL, Shende DZ. Reactive extraction of acrylic acid with tri-n-butyl phosphate in natural oils. J Chem Technol Biotechnol 2017;92(11):2825-34; doi: 10.1002/ jctb.5295

10. Bhalla TC, Kumar V, Kumar V, Thakur N. Nitrile metabolizing enzymes in biocatalysis and biotransformation. Appl Biochem Biotechnol 2018;185(4):925-46; doi: 10.1007/s12010-018-2705-7

11. Gong JS, Shi JS, Lu ZM, Li H, Zhou ZM, Xu ZH. Nitrile-converting enzymes as a tool to improve biocatalysis in organic synthesis: recent insights and promises. Crit Rev Biotechnol 2017;37(1):69-81; doi: 10.3109/07388551.2015.1120704

12. Wu Z, Liu C, Zhang Z, Zheng R, Zheng Y. Amidase as a versatile tool in amide-bond cleavage: From molecular features to biotechnological applications. Biotechnol Adv 2020;43:107574; doi: 10.1016/j. biotechadv.2020.107574

13. Lin MM. Selective oxidation of propane to acrylic acid with molecular oxygen. Appl Catal A Gen 2001;207(1-2):1-6; doi: 10.1016/S0926- 860X(00)00609-8

14. Sun J, Yu H, Chen J, Luo H, Shen Z. Ammonium acrylate biomanufacturing by an engineered Rhodococcus ruber with nitrilase overexpression and double-knockout of nitrile hydratase and amidase. J Ind Microbiol Biotechnol 2016;43(12):1631-9; doi: 10.1007/ s10295-016-1840-9

15. Gong JS, Lu ZM, Li H, Shi JS, Zhou ZM, Xu ZH. Nitrilases in nitrile biocatalysis: recent progress and forthcoming research. Microb Cell Fact 2012;11(1):1-8; doi: 10.1186/1475-2859-11-142

16. Wasewar KL. Reactive extraction: an intensifying approach for carboxylic acid separation. Int J Chem Eng Appl 2012;3(4):249; doi: 10.7763/IJCEA.2012.V3.195

17. Wasewar KL, Shende D, Keshav A. Reactive extraction of itaconic acid using tri-n-butyl phosphate and aliquat 336 in sunflower oil as a non-toxic diluent. J Chem Technol Biotechnol 2011;86(2):319-23; doi: 10.1002/jctb.2500

18. Keshav A, Wasewar KL, Chand S. Extraction of propionic acid with tri-n-octyl amine in different diluents. Sep Purif Technol 2008;63(1):179- 83; doi: 10.1016/j.seppur.2008.04.012

19. Zhou X, Liu L, Chen Y, Xu S, Chen J. Efficient biodegradation of cyanide and ferrocyanide by Na-alginate beads immobilized with fungal cells of Trichoderma koningii. Can J Microbiol 2007;53(9):1033-7; doi: 10.1139/W07-070

20. Arfi T, Nigam VK. Extraction and quantification of nicotinic acid from 3-cyanopyrdinase catalysed reaction system. Res J Chem Environ 2018; 22 (9): 7-13.

21. Prabha R, Nigam VK. Biotransformation of acrylamide to acrylic acid carried through acrylamidase enzyme synthesized from whole cells of Bacillus tequilensis (BITNR004). Biocatal Biotransfor 2020;38(6):445-56; doi: 10.1080/10242422.2020.1780211

22. Pacheco R, Karmali A, Serralheiro ML, Haris PI. Application of fourier transform infrared spectroscopy for monitoring hydrolysis and synthesis reactions catalyzed by a recombinant amidase. Anal Biochem 2005;346(1):49-58; doi: 10.1016/j.ab.2005.07.027

23. Pacheco R, Serralheiro ML, Karmali A, Haris PI. Measuring enzymatic activity of a recombinant amidase using fourier transform infrared spectroscopy. Anal Biochem 2003;322(2):208-14; doi: 10.1016/j. ab.2003.07.012

24. Kolev T, Spiteller M, Koleva B. Spectroscopic and structural elucidation of amino acid derivatives and small peptides: experimental and theoretical tools. Amino Acids 2010;38(1):45-50; doi: 10.1007/ s00726-008-0220-9

25. Rubin-Bejerano IF, Fink GR, Osler H. Canadian patents database/ patent 2760044 summary. Canadian Intellectual Property Office, Gatineau, Canada, 2010.

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