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A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates

Published in ISI journal. Full text available in SpringerLink . http://link.springer.com/article/10.1007%2Fs12257-012-0607-z

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Main Authors: R. Zuriani, S. Vigneswari, M. N. M. Azizan, M. I. A. Majid, A. A. Amirul, UniKL Chan
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Published: Springer Berlin Heidelberg 2014
Subjects:
polyhydroxyalkanoate
nile red staining
excitation and emission wavelength
fluorescence intensity
Online Access:http://localhost/xmlui/handle/123456789/7201
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spelling my.unikl.ir-72012014-07-14T08:05:10Z A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates R. Zuriani S. Vigneswari M. N. M. Azizan M. I. A. Majid A. A. Amirul UniKL Chan polyhydroxyalkanoate nile red staining excitation and emission wavelength fluorescence intensity Published in ISI journal. Full text available in SpringerLink . http://link.springer.com/article/10.1007%2Fs12257-012-0607-z A rapid quantitative measurement of accumulated polyhydroxyalkanoate (PHA) is essential for rapid monitoring of PHA production by microorganisms. In the present study, a 96-well microplate was used as a high throughput means to measure the fluorescence intensity of the Nile red stained cells containing PHA. The linear correlation obtained between intracellular PHA concentration and the fluorescence intensity represents the potential of the Nile red method employment to determine PHA concentration. The optimal ranges of excitation and emission wavelengths were determined using bacterial cells containing different types of PHAs, of different co-monomers and compositions. Interestingly, in spite of different co-monomers compositions in each PHA, all tested PHAs fluoresced maximally at excitation wavelength between 520 and 550 nm, and emission wavelength between 590 and 630 nm. The developed staining method also had successfully demonstrated a good correlation between the amount of accumulated PHA based on the fluorescence intensity measurements and that from chromatographic analysis to evaluate poly(3-hydroxybutyrate) [P(3HB)], poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)], poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] and poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) [P(3HB-co-3HV-co-4HB)], using the same calibration curve, despite of different co-monomers that the PHA consist. Strongly supported by these experimental results, it can therefore be concluded that the developed staining method can be efficiently applied for rapid monitoring of PHA production. 2014-07-14T08:05:10Z 2014-07-14T08:05:10Z 2013 Zuriani, R., Vigneswari, S., Azizan, M. N. M., Majid, M. I. A., & Amirul, A. A. (2013). A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates. Biotechnology and Bioprocess Engineering, 18(3), 472–478. doi:10.1007/s12257-012-0607-z 1976-3816 http://localhost/xmlui/handle/123456789/7201 Biotechnology and Bioprocess Engineering; Springer Berlin Heidelberg
institution Universiti Kuala Lumpur
building UniKL Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Kuala Lumpur
content_source UniKL Institutional Repository
url_provider http://ir.unikl.edu.my/
topic polyhydroxyalkanoate
nile red staining
excitation and emission wavelength
fluorescence intensity
spellingShingle polyhydroxyalkanoate
nile red staining
excitation and emission wavelength
fluorescence intensity
R. Zuriani
S. Vigneswari
M. N. M. Azizan
M. I. A. Majid
A. A. Amirul
UniKL Chan
A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates
description Published in ISI journal. Full text available in SpringerLink . http://link.springer.com/article/10.1007%2Fs12257-012-0607-z
format
author R. Zuriani
S. Vigneswari
M. N. M. Azizan
M. I. A. Majid
A. A. Amirul
UniKL Chan
author_facet R. Zuriani
S. Vigneswari
M. N. M. Azizan
M. I. A. Majid
A. A. Amirul
UniKL Chan
author_sort R. Zuriani
title A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates
title_short A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates
title_full A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates
title_fullStr A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates
title_full_unstemmed A high throughput Nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates
title_sort high throughput nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates
publisher Springer Berlin Heidelberg
publishDate 2014
url http://localhost/xmlui/handle/123456789/7201
_version_ 1644484883511246848
score 13.211869

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