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An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains
Cytidine is an industrially useful precursor for the production of antiviral compounds and a variety of industrial compounds. Interest in the microbial production of cytidine has grown recently and high-throughput screening of cytidine over-producers is an important approach in large-scale industria...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4376533/ https://www.ncbi.nlm.nih.gov/pubmed/25816248 http://dx.doi.org/10.1371/journal.pone.0121612 |
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author | Dong, Huina Liu, Yongfei Zu, Xin Li, Ning Li, Feiran Zhang, Dawei |
author_facet | Dong, Huina Liu, Yongfei Zu, Xin Li, Ning Li, Feiran Zhang, Dawei |
author_sort | Dong, Huina |
collection | PubMed |
description | Cytidine is an industrially useful precursor for the production of antiviral compounds and a variety of industrial compounds. Interest in the microbial production of cytidine has grown recently and high-throughput screening of cytidine over-producers is an important approach in large-scale industrial production using microorganisms. An enzymatic assay for cytidine was developed combining cytidine deaminase (CDA) and indophenol method. CDA catalyzes the cleavage of cytidine to uridine and NH(3), the latter of which can be accurately determined using the indophenol method. The assay was performed in 96-well plates and had a linear detection range of cytidine of 0.058 - 10 mM. This assay was used to determine the amount of cytidine in fermentation flasks and the results were compared with that of High Perfomance Liquid Chromatography (HPLC) method. The detection range of the CDA method is not as wide as that of the HPLC, furthermore the correlation factor of CDA method is not as high as that of HPLC. However, it was suitable for the detection of large numbers of crude samples and was applied to high-throughput screening for high cytidine-producing strains using 96-well deep-hole culture plates. This assay was proved to be simple, accurate, specific and suitable for cytidine detection and high-throughput screening of cytidine-producing strains in large numbers of samples (96 well or more). |
format | Online Article Text |
id | pubmed-4376533 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-43765332015-04-04 An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains Dong, Huina Liu, Yongfei Zu, Xin Li, Ning Li, Feiran Zhang, Dawei PLoS One Research Article Cytidine is an industrially useful precursor for the production of antiviral compounds and a variety of industrial compounds. Interest in the microbial production of cytidine has grown recently and high-throughput screening of cytidine over-producers is an important approach in large-scale industrial production using microorganisms. An enzymatic assay for cytidine was developed combining cytidine deaminase (CDA) and indophenol method. CDA catalyzes the cleavage of cytidine to uridine and NH(3), the latter of which can be accurately determined using the indophenol method. The assay was performed in 96-well plates and had a linear detection range of cytidine of 0.058 - 10 mM. This assay was used to determine the amount of cytidine in fermentation flasks and the results were compared with that of High Perfomance Liquid Chromatography (HPLC) method. The detection range of the CDA method is not as wide as that of the HPLC, furthermore the correlation factor of CDA method is not as high as that of HPLC. However, it was suitable for the detection of large numbers of crude samples and was applied to high-throughput screening for high cytidine-producing strains using 96-well deep-hole culture plates. This assay was proved to be simple, accurate, specific and suitable for cytidine detection and high-throughput screening of cytidine-producing strains in large numbers of samples (96 well or more). Public Library of Science 2015-03-27 /pmc/articles/PMC4376533/ /pubmed/25816248 http://dx.doi.org/10.1371/journal.pone.0121612 Text en © 2015 Dong et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Dong, Huina Liu, Yongfei Zu, Xin Li, Ning Li, Feiran Zhang, Dawei An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains |
title | An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains |
title_full | An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains |
title_fullStr | An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains |
title_full_unstemmed | An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains |
title_short | An Enzymatic Assay for High-Throughput Screening of Cytidine-Producing Microbial Strains |
title_sort | enzymatic assay for high-throughput screening of cytidine-producing microbial strains |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4376533/ https://www.ncbi.nlm.nih.gov/pubmed/25816248 http://dx.doi.org/10.1371/journal.pone.0121612 |
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