Optimization of methodology for EPS extraction and analysis

The chemical reagents ‘NaCl+HCHO’ are often used as an extractant for EPS extraction due to the neutral pH. However, there is inadequate discussion of the reagent in the literature. In this study, an extraction process using ‘NaCl+HCHO’ was examined and optimized via an investigation of the relation...

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Main Author: Tan, Shu Hui.
Other Authors: Liu Yu
Format: Final Year Project
Language:English
Published: 2010
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Online Access:http://hdl.handle.net/10356/39532
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-395322023-03-03T16:51:20Z Optimization of methodology for EPS extraction and analysis Tan, Shu Hui. Liu Yu School of Civil and Environmental Engineering DRNTU::Engineering::Environmental engineering::Water treatment The chemical reagents ‘NaCl+HCHO’ are often used as an extractant for EPS extraction due to the neutral pH. However, there is inadequate discussion of the reagent in the literature. In this study, an extraction process using ‘NaCl+HCHO’ was examined and optimized via an investigation of the relationship between the extraction solution, centrifuge speed, ultrasound configurations and biomass concentration with the yield of extraction. Comparisons were made with extraction by ‘NaOH+HCHO’, which was found in the literature as the most effective chemical reagent for extraction to date. The Lowry and Bradford assays for protein measurement was also studied and compared. It was found that low centrifuge speed of 2000 rpm and 4000 rpm will cause biomass re-suspension. Sonication at 15 W/ml and above at 50 kHz for more than 1.5 min might lead to cell lysis. ‘NaCl+HCHO’ will only be effective at extracting biomass below 20mg. The optimized ‘NaCl+HCHO’ extraction process will use 6000rpm for centrifugation and sonication at 12.5W/ml at 50Hz for 1min. However, ‘NaOH+HCHO’ process was still more effective than the optimized ‘NaCl+HCHO’ process. Lowry assay was more effective and sensitive then Bradford assay in the range 0mg/l to 200mg/l proteins. However Lowry assay was prone to interferences and require long experimental time. Bachelor of Engineering (Environmental Engineering) 2010-05-31T01:12:51Z 2010-05-31T01:12:51Z 2010 2010 Final Year Project (FYP) http://hdl.handle.net/10356/39532 en Nanyang Technological University 39 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Engineering::Environmental engineering::Water treatment
spellingShingle DRNTU::Engineering::Environmental engineering::Water treatment
Tan, Shu Hui.
Optimization of methodology for EPS extraction and analysis
description The chemical reagents ‘NaCl+HCHO’ are often used as an extractant for EPS extraction due to the neutral pH. However, there is inadequate discussion of the reagent in the literature. In this study, an extraction process using ‘NaCl+HCHO’ was examined and optimized via an investigation of the relationship between the extraction solution, centrifuge speed, ultrasound configurations and biomass concentration with the yield of extraction. Comparisons were made with extraction by ‘NaOH+HCHO’, which was found in the literature as the most effective chemical reagent for extraction to date. The Lowry and Bradford assays for protein measurement was also studied and compared. It was found that low centrifuge speed of 2000 rpm and 4000 rpm will cause biomass re-suspension. Sonication at 15 W/ml and above at 50 kHz for more than 1.5 min might lead to cell lysis. ‘NaCl+HCHO’ will only be effective at extracting biomass below 20mg. The optimized ‘NaCl+HCHO’ extraction process will use 6000rpm for centrifugation and sonication at 12.5W/ml at 50Hz for 1min. However, ‘NaOH+HCHO’ process was still more effective than the optimized ‘NaCl+HCHO’ process. Lowry assay was more effective and sensitive then Bradford assay in the range 0mg/l to 200mg/l proteins. However Lowry assay was prone to interferences and require long experimental time.
author2 Liu Yu
author_facet Liu Yu
Tan, Shu Hui.
format Final Year Project
author Tan, Shu Hui.
author_sort Tan, Shu Hui.
title Optimization of methodology for EPS extraction and analysis
title_short Optimization of methodology for EPS extraction and analysis
title_full Optimization of methodology for EPS extraction and analysis
title_fullStr Optimization of methodology for EPS extraction and analysis
title_full_unstemmed Optimization of methodology for EPS extraction and analysis
title_sort optimization of methodology for eps extraction and analysis
publishDate 2010
url http://hdl.handle.net/10356/39532
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