| ORIGINAL ARTICLE |
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| Year : 2022 | Volume
: 11
| Issue : 1 | Page : 110 |
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Expression optimizing of recombinant Oxalyl-CoA decarboxylase in Escherichia coli
Fatemeh Abarghooi Kahaki1, Seyed Mohsen Dehnavi2
1 Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
2 Department of Cell and Molecular Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran
Correspondence Address:
Dr. Seyed Mohsen Dehnavi
Shahid Beheshti University, Daneshjoo Boulevard, Velenjak, Tehran
Iran
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/abr.abr_244_21
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Background: One of the most common diseases of the urinary tract is stones of this system, including kidney stones. About 70%–80% of kidney stones are calcium oxalate. Oxalyl-CoA decarboxylase is a single polypeptide included of 568 amino acids which play a key role in oxalate degradation.
Materials and Methods: The aim of current study is high-level expression of oxalyl-CoA decarboxylase in Escherichia coli BL21 (DE3). To achieve this aim, oxalyl-CoA decarboxylase gene was cloned upon pET-30a (+) with T7 promoter. The vector containing the oxalyl-CoA decarboxylase gene was transformed into E. coli and the expression of the gene was examined on a laboratory scale and fermentor. At first, the effect of temperature, culture medium, and induction time on oxalyl-CoA decarboxylase expression at three levels was examined.
Results: The obtained data showed that the highest expression was related to the terrific broth culture medium and temperature of 32°C with an inducer concentration of 1 mM. Under this situation the ultimate cells dry weight and the final oxalyl-CoA decarboxylase expression were 2.46 g/l and 36% of total protein, respectively. Then induction time was optimized in a bench bioreactor and productivity of oxalyl-CoA decarboxylase was calculated. Under optimized condition the cell density, biomass productivity and oxalyl-CoA decarboxylase concentration reached 4.02 g/l, 0.22 g/l/h, and 0.7 g/l which are one of the highest reported rates.
Conclusion: This study demonstrated that high levels of oxalyl-CoA decarboxylase can be achieved by optimizing the expression conditions.
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