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Expression of the gene encoding oxalate decarboxylase from Bacillus subtilis and characterization of the recombinant enzyme
BACKGROUND: The concentration of urinary oxalate is more influential to the formation of calcium oxalate urolithiasis than is urinary calcium concentration. YvrK gene encodes a 43 KD-sized oxalate decarboxylase. We previously developed the recombinant Escherichia coli (E. coli) expressing Yvrk gene...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4161865/ https://www.ncbi.nlm.nih.gov/pubmed/25186982 http://dx.doi.org/10.1186/1756-0500-7-598 |
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author | Lee, Eunhye Jeong, Byong Chang Park, Yong Hyun Kim, Hyeon Hoe |
author_facet | Lee, Eunhye Jeong, Byong Chang Park, Yong Hyun Kim, Hyeon Hoe |
author_sort | Lee, Eunhye |
collection | PubMed |
description | BACKGROUND: The concentration of urinary oxalate is more influential to the formation of calcium oxalate urolithiasis than is urinary calcium concentration. YvrK gene encodes a 43 KD-sized oxalate decarboxylase. We previously developed the recombinant Escherichia coli (E. coli) expressing Yvrk gene from Bacillus subtilis and named it as pBy. The aim of this study was to purify the recombinant oxalate decarboxylase overexpressed in E. coli and evaluate the oxalate-degrading activity of the purified enzyme. RESULTS: The oxalate-degrading activity of pBy was highest when cultured at pH 5. The activity of purified oxalate decarboxylase was determined after incubation with sodium oxalate and the optimal pH and temperature of oxalate decarboxylase were determined. Purified oxalate decarboxylase degraded more than 50% of oxalate when incubated with MnCl(2) and sodium oxalate in atmospheric O(2). The optimal pH of recombinant oxalate decarboxylase was 5 and the optimal temperature was 28°C. Eight-week-old Sprague–Dawley male rats were used as a transient hyperoxaluric rat model. Suprapubic catheter was inserted into the bladder of each rat and urine was collected hourly before and 3 hours after oral oxalate intake in the absence and presence of homogenates of pBy and non-recombinant E. coli as the control. After the oral intake of sodium oxalate, the concentration of oxalate in urine increased exponentially for 3 hours. The oxalate concentration in urine was decreased significantly by pBy homogenates compared to control. CONCLUSIONS: We constructed the recombinant E. coli expressing YvrK gene and purified the recombinant oxalate decarboxylase successfully. Purified recombinant oxalate decarboxylase, as well as recombinant E. coli named pBy, showed the oxalate-degrading activity in in vitro and in vivo model. |
format | Online Article Text |
id | pubmed-4161865 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-41618652014-09-13 Expression of the gene encoding oxalate decarboxylase from Bacillus subtilis and characterization of the recombinant enzyme Lee, Eunhye Jeong, Byong Chang Park, Yong Hyun Kim, Hyeon Hoe BMC Res Notes Research Article BACKGROUND: The concentration of urinary oxalate is more influential to the formation of calcium oxalate urolithiasis than is urinary calcium concentration. YvrK gene encodes a 43 KD-sized oxalate decarboxylase. We previously developed the recombinant Escherichia coli (E. coli) expressing Yvrk gene from Bacillus subtilis and named it as pBy. The aim of this study was to purify the recombinant oxalate decarboxylase overexpressed in E. coli and evaluate the oxalate-degrading activity of the purified enzyme. RESULTS: The oxalate-degrading activity of pBy was highest when cultured at pH 5. The activity of purified oxalate decarboxylase was determined after incubation with sodium oxalate and the optimal pH and temperature of oxalate decarboxylase were determined. Purified oxalate decarboxylase degraded more than 50% of oxalate when incubated with MnCl(2) and sodium oxalate in atmospheric O(2). The optimal pH of recombinant oxalate decarboxylase was 5 and the optimal temperature was 28°C. Eight-week-old Sprague–Dawley male rats were used as a transient hyperoxaluric rat model. Suprapubic catheter was inserted into the bladder of each rat and urine was collected hourly before and 3 hours after oral oxalate intake in the absence and presence of homogenates of pBy and non-recombinant E. coli as the control. After the oral intake of sodium oxalate, the concentration of oxalate in urine increased exponentially for 3 hours. The oxalate concentration in urine was decreased significantly by pBy homogenates compared to control. CONCLUSIONS: We constructed the recombinant E. coli expressing YvrK gene and purified the recombinant oxalate decarboxylase successfully. Purified recombinant oxalate decarboxylase, as well as recombinant E. coli named pBy, showed the oxalate-degrading activity in in vitro and in vivo model. BioMed Central 2014-09-03 /pmc/articles/PMC4161865/ /pubmed/25186982 http://dx.doi.org/10.1186/1756-0500-7-598 Text en © Lee et al.; licensee BioMed Central Ltd. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Lee, Eunhye Jeong, Byong Chang Park, Yong Hyun Kim, Hyeon Hoe Expression of the gene encoding oxalate decarboxylase from Bacillus subtilis and characterization of the recombinant enzyme |
title | Expression of the gene encoding oxalate decarboxylase from Bacillus subtilis and characterization of the recombinant enzyme |
title_full | Expression of the gene encoding oxalate decarboxylase from Bacillus subtilis and characterization of the recombinant enzyme |
title_fullStr | Expression of the gene encoding oxalate decarboxylase from Bacillus subtilis and characterization of the recombinant enzyme |
title_full_unstemmed | Expression of the gene encoding oxalate decarboxylase from Bacillus subtilis and characterization of the recombinant enzyme |
title_short | Expression of the gene encoding oxalate decarboxylase from Bacillus subtilis and characterization of the recombinant enzyme |
title_sort | expression of the gene encoding oxalate decarboxylase from bacillus subtilis and characterization of the recombinant enzyme |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4161865/ https://www.ncbi.nlm.nih.gov/pubmed/25186982 http://dx.doi.org/10.1186/1756-0500-7-598 |
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