Volume 8, Issue 3 (Journal of Clinical and Basic Research (JCBR) 2024)
jcbr 2024, 8(3): 10-13 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Shahbazmohammadi H. Response surface optimization for lactose inducible expression of recombinant fructosyl peptide oxidase enzyme in Escherichia coli. jcbr 2024; 8 (3) :10-13
URL: http://jcbr.goums.ac.ir/article-1-454-en.html
URL: http://jcbr.goums.ac.ir/article-1-454-en.html
Cellular and Molecular Research Center, Research Institute for Prevention of Non-Communicable Diseases, Qazvin University of Medical Sciences, Qazvin, Iran , sh_hamid_biochem@yahoo.com
Abstract: (322 Views)
Background: Fructosyl peptide oxidase (FPOX), a flavoenzyme classified as an oxidoreductase, serves as a diagnostic enzyme in HbA1c measurement tests. This research focuses on statistically optimizing lactose-induced expression to produce soluble recombinant FPOX.
Methods: A Plackett–Burman design was used to identify key factors influencing enzyme expression, which were further optimized using a central composite design.
Results: The results indicated that glycerol, yeast extract, tryptone, and lactose significantly affected FPOX activity. The maximum enzyme activity and biomass concentration were achieved under the optimum conditions of yeast extract (10.12 g/L), tryptone (13.44 g/L), K₂HPO₄ (2.62 g/L), and lactose (12.79 g/L). When the lactose-inducible induction strategy was examined at the shake flask scale, FPOX activity (28.77 U/mL) was 18.5-fold higher than with the IPTG induction protocol. Additionally, the increased biomass yield (49.0 g/L compared to 22.0 g/L) further supported the appropriateness of utilizing lactose-inducible expression.
Conclusion: Together, our findings indicated that the design of experiment methodology can be utilized effectively to enhance the production of the FPOX enzyme with lactose as the inducer.
Methods: A Plackett–Burman design was used to identify key factors influencing enzyme expression, which were further optimized using a central composite design.
Results: The results indicated that glycerol, yeast extract, tryptone, and lactose significantly affected FPOX activity. The maximum enzyme activity and biomass concentration were achieved under the optimum conditions of yeast extract (10.12 g/L), tryptone (13.44 g/L), K₂HPO₄ (2.62 g/L), and lactose (12.79 g/L). When the lactose-inducible induction strategy was examined at the shake flask scale, FPOX activity (28.77 U/mL) was 18.5-fold higher than with the IPTG induction protocol. Additionally, the increased biomass yield (49.0 g/L compared to 22.0 g/L) further supported the appropriateness of utilizing lactose-inducible expression.
Conclusion: Together, our findings indicated that the design of experiment methodology can be utilized effectively to enhance the production of the FPOX enzyme with lactose as the inducer.
Article Type: Research |
Subject:
Biochemistry
References
1. Ogawa N, Kimura T, Umehara F, Katayama Y, Nagai G, Suzuki K, et al. Creation of haemoglobin A1c direct oxidase from fructosyl peptide oxidase by combined structure based site specific mutagenesis and random mutagenesis. Sci Rep. 2019;9(1):942. [View at Publisher] [DOI] [PMID] [Google Scholar]
2. Shahbazmohammadi H, Sardari S, Lari A, Omidinia E. Engineering an efficient mutant of Eupenicillium terrenum fructosyl peptide oxidase for the specific determination of hemoglobin A1c. Appl Microbiol Biotechnol. 2019;103(4):1725-35. [View at Publisher] [DOI] [PMID] [Google Scholar]
3. Chen KJ, Wang CH, Liao CW, Lee CK. Recombinant fructosyl peptide oxidase preparation and its immobilization on polydopamine coating for colorimetric determination of HbA1c. Int J Biol Macromol. 2018;120(Pt A):325-31. [View at Publisher] [DOI] [PMID] [Google Scholar]
4. Hsieh KM, Lan KC, Hua WL, Chen MK, Jang LS, Wang MH. Glycated hemoglobin (HbA1c) affinity biosensors withring-shaped interdigital electrodes on impedance measurement. Biosens Bioelectron. 2013;49:450-6. [View at Publisher] [DOI] [PMID] [Google Scholar]
5. Kim S, Miura S, Ferri S, Tsugawa W, Sode K. Cumulative effect of amino acid substitution for the development of fructosyl valine-specific fructosyl amine oxidase. Enzyme Microb Technol. 2009;44(1):52-6. [View at Publisher] [DOI] [Google Scholar]
6. Miura S, Ferri S, Tsugawa W, Kim S, Sode K. Development of fructosyl amine oxidase specific to fructosyl valine by site-directed mutagensis. Protein Eng Des Sel. 2008;21(4):233-9. [View at Publisher] [DOI] [PMID] [Google Scholar]
7. Hatada M, Saito S, Yonehara S, Tsugawa W, Asano R, Ikebukuro K, et al. Development of glycated peptide enzyme sensor based flow injection analysis system for haemoglobin A1c monitoring using quasi-direct electron transfer type engineered fructosyl peptide oxidase. Biosens Bioelectron. 2021;177:112984. [View at Publisher] [DOI] [PMID] [Google Scholar]
8. Singh P, Singh Shera S, Banik J, Mohan Banik R. Optimization of cultural conditions using response surface methodology versus artificial neural network and modeling of L-glutaminase production by Bacillus cereus MTCC 1305. Bioresour Technol. 2013;137:261-9. [View at Publisher] [DOI] [PMID] [Google Scholar]
9. Shahbaz Mohammadi H, Mostafavi SS, Soleimani S, Bozorgian S, Pooraskari M, Kianmehr A. Response surface methodology to optimize partition and purification of two recombinant oxidoreductase enzymes, glucose dehydrogenase and D-galactose dehydrogenase in aqueous two-phase systems. Protein Expression Purifi. 2015;108:41-7. [View at Publisher] [DOI] [PMID] [Google Scholar]
10. Khayet M, Cojocaru C, Zakrzewska-Trznadel G. Response surface modeling and optimization in pervaporation. J Membr Sci. 2008;321(2):272-83. [View at Publisher] [DOI] [Google Scholar]
11. Ashrafi-Saiedlou S, Rasouli-Sadaghiani M, Samadi A, Barin M, Sepehr E. Aspergillus niger as an eco-friendly agent for potassium release from K- bearing minerals: isolation, screening and culture medium optimization using Plackett-Burman design and response surface methodology. Heliyon. 2024;10(7):e29117. [View at Publisher] [DOI] [PMID] [Google Scholar]
12. Moore B, Georgakis C, Antoniou C, Khattak S. A two-phase approach optimizing productivity for a mAb-producing CHO cell culture process using dynamic response surface methodology models. Biochem Eng J. 2024;201:109137. [View at Publisher] [DOI] [Google Scholar]
13. Abdullah R, Ahmad S, Nisar K, Kaleem A, Iqtedar M. Response surface methodology as an approach for optimization of alpha amylase production by using bacterial consortium under submerged fermentation. Kuwait J Sci. 2024;51(3):100220. [View at Publisher] [DOI] [Google Scholar]
14. Mutanda I, Zahoor, Sethupathy S, Xu Q, Zhu B, Shah SWA, et al. Optimization of heterologous production of Bacillus ligniniphilus L1 laccase in Escherichia coli through statistical design of experiments. Microbiol Res. 2023;274:127416. [View at Publisher] [DOI] [PMID] [Google Scholar]
15. Wardah ZH, Chaudhari HG, Prajapati V, Raol GG. Application of statistical methodology for the optimization of l-glutaminase enzyme production from Streptomyces pseudogriseolus ZHG20 under solid-state fermentation. J Genet Eng Biotechnol. 2023;21(1):138. [View at Publisher] [DOI] [PMID] [Google Scholar]
16. Sharma P, Sharma N. RSM approach to pre-treatment of lignocellulosic waste and a statistical methodology for optimizing bioethanol production. Waste Manage Bull. 2024;2(1):49-66. [View at Publisher] [DOI] [Google Scholar]
17. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A laboratory manual. 2nd ed. New York:Cold Spring Harbor Laboratory press;1989. [View at Publisher] [Google Scholar]
18. Shahbazmohammadi H, Omidinia E. Medium optimization for improved production of dihydrolipohyl dehydrogenase from Bacillus sphaericus PAD-91 in Escherichia coli. Mol Biotechnol. 2017;59(7):260-70. [View at Publisher] [DOI] [PMID] [Google Scholar]
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0).