The stability increase of α-amylase enzyme from Aspergillus fumigatus using dimethyladipimidate
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Yandri Yandri
,
Nurmala Nurmala
Abstract
This study’s purpose is to improve the α-amylase enzyme’s stability from Aspergillus fumigatus applying dimethyladipimidate (DMA). It was conducted in different stages, including production, isolation, purification, modification, and the characterization of native and modified enzymes by the DMA addition. The enzyme activity was specified using the Fuwa and Mandels methods, while the protein level was conducted by the Lowry method. The results indicated that the native enzyme contains a specific activity of 7010.42 U/mg, with an increase of 7.8 times than the crude extract, which contains 904.38 U/mg. Meanwhile, the native enzyme contains an optimum pH of 5 at 55 °C, with residual activity of 17.17% after 60 min of incubation at 55 °C and a half-life of 25.86 min. After the DMA addition with 0.5, 1, and 1.5% concentration, the enzymes had 5.5 optimum pH and 65 °C temperature. Meanwhile, after 60 min of incubation at 65 °C, the modified enzymes had 54.17, 46.18, and 34.44% of residual activity, and 85.55 58.25 and 37.46 min of half-lives, respectively. This showed that the addition of DMA to the native α-amylase from A. fumigatus increased the stability of the modified enzymes by 1.5–3.3 times than the native enzyme.
Funding source: Direktorat Jenderal Pendidikan Tinggi and Ministry of Education, Culture, Research, and Technology
Award Identifier / Grant number: 032/E4.1/AK.04.PT/2021, 12 July 2021
Award Identifier / Grant number: 3972/UN26.21/PN/2021, 14 July 2021
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Author contribution: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
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Research funding: This study was supported by Ministry of Education, Culture, Research, and Technology through Basic Research 2021 number 032/E4.1/AK.04.PT/2021, 12 July 2021 and 3972/UN26.21/PN/2021, 14 July 2021.
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Conflict of interest statement: The authors declare no conflicts of interest regarding this article.
References
1. Li, S, Yang, X, Yang, S, Zhu, M, Wang, X. Technology prospecting on enzymes: application, marketing and engineering. Comput Struct Biotechnol J 2012;2:1–11. https://doi.org/10.5936/csbj.201209017.Search in Google Scholar PubMed PubMed Central
2. Sirisha, VL, Jain, A, Jain, A. Enzyme immobilization: an overview on methods, support material, and applications of immobilized enzymes. Adv Food Nutr Res 2016;79:179–211. https://doi.org/10.1016/bs.afnr.2016.07.004.Search in Google Scholar PubMed
3. Yandri, Suhartati, T, Hadi, S. Immobilization of α-amylase from locale bacteria isolate Bacillus subtilis ITBCCB148 with diethylaminoethyl cellulose (DEAE-Cellulose). Mat Sci Res India 2010;7:123–8. https://doi.org/10.13005/msri/070114.Search in Google Scholar
4. Yandri, Susanti, D, Suhartati, T, Hadi, S. Immobilization of α-amylase from locale bacteria isolate Bacillus subtilis ITBCCB148 with carboxymethyl cellulose (CM-Cellulose). Mod Appl Sci 2012;6:81–6. https://doi.org/10.5539/mas.v6n3p81.Search in Google Scholar
5. Yandri, Amalia, P, Suhartati, T, Hadi, S. Effect of immobilization towards thermal stability of α-amylase isolates from locale bacteria isolate Bacillus subtilis ITBCCB148 with calcium alginate. Asian J Chem 2013;25:6897–9. https://doi.org/10.14233/ajchem.2013.15111.Search in Google Scholar
6. Yandri, Suhartati, T, Yuwono, SD, Qudus, HI, Tiarsa, ER, Hadi, S. value="α-[?show $132#?]amylase"Immobilization of α-amylase from Bacillus subtilis ITBCCB148 using bentonit. Asian J Microbiol Biotechnol Environ Sci 2018;20:487–92.Search in Google Scholar
7. Yandri, Suhartati, T, Satria, H, Widyasmara, A, Hadi, S. Increasing stability of α-amylase obtained from Bacillus subtilis ITBCCB148 by immobilization with chitosan. Mediterr J Chem 2020;10:155–61. https://doi.org/10.13171/mjc10202002131126ysh.Search in Google Scholar
8. Sarrouh, B, Santos, TM, Miyoshi, A, Dias, R, Azevedo, V. Up-to-date insight on industrial enzymes applications and global market. J Bioprocess Biotech 2012;S4:002. https://doi.org/10.4172/2155-9821.s4-002.Search in Google Scholar
9. Yandri, Y, Tiarsa, ER, Suhartati, Satria, H, Irawan, B, Hadi, S. The stability improvement of α-amylase enzyme from Aspergillus fumigatus by immobilization on a bentonite matrix. Biochem Res Int 2022;2022:3797629. https://doi.org/10.1155/2022/3797629.Search in Google Scholar PubMed PubMed Central
10. Vieille, C, Zeikus, JG. Thermozymes: identifying molecular determinant of protein structural and functional stability. Trends Biotech 1996;14:183–9. https://doi.org/10.1016/0167-7799(96)10026-3.Search in Google Scholar
11. Mozhaev, VV, Martinek, K. Structure-stability relationship in proteins: new approaches to stabilizing enzymes. Enzym Microb Technol 1984;6:50–9. https://doi.org/10.1016/0141-0229(84)90034-6.Search in Google Scholar
12. Janecek, S. Strategies for obtaining stable enzymes. Process Biochem 1993;28:435–45.10.1016/0032-9592(93)85026-CSearch in Google Scholar
13. Tananchai, P. Stabilization of enzymes by chemical modification [Thesis]. New Zealand: Massey University; 2011.Search in Google Scholar
14. Elangovan, H. Chemical cross-linking protein: a review. J Pharmaceut Sci Innovat 2012;1:22–6.Search in Google Scholar
15. Wong, SS, Jameson, DM. Chemistry of protein and nucleic acid cross-linking and conjugation. Boca Raton: Taylor & Francis. United States of America; 2012.10.1201/b11175Search in Google Scholar
16. Yandri, Amalia, P, Suhartati, T, Hadi, S. The chemical modification of cellulase obtained from Bacillus subtilis ITBCCB148 with dimethyladimipidate. Biosci Biotechnol Res Asia 2015;12:2089–93. https://doi.org/10.13005/bbra/1877.Search in Google Scholar
17. Yandri, A, Apriyanti, Suhartati, T, Hadi, S. The increase of thermal stability of α- amylase from locale bacteria Isolate Bacillus subtilis ITBCCB148 by chemical modification with dimethyladipimidate. Biosci Biotechnol Res Asia 2010;7:713–8. https://doi.org/10.13005/msri/070114.Search in Google Scholar
18. Mandels, M, Raymond, R. Measurement of saccharifying cellulose. Biotechnol Bioeng Symp 1976;6:21–33.Search in Google Scholar
19. Lowry, OH, Rosebrough, NJ, Farr, AL, Randall, RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951;193:265–75. https://doi.org/10.1016/s0021-9258(19)52451-6.Search in Google Scholar
20. Kazan, D, Ertan, H, Erarslan, A. Stabilization of Escherichia coli penicillin g acylase against thermal inactivation by cross-linking with dextran dialdehyde polymers. Appl Microbiol Biotechnol 1997;48:191–7. https://doi.org/10.1007/s002530051037.Search in Google Scholar PubMed
21. Stahl, S. In: Gupta, MN, editor. Thermostability of Enzymes. New Delhi: Springer-Verlag; 1999:59–60 pp.Search in Google Scholar
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