Optimization of glycerol fermentation conditions for bioethanol production by locally isolated Escherichia coli SS1

The increase of biodiesel production worldwide lead to the increase of glycerol present in environment. Bioconverting glycerol into various valuable products is one of glycerol’s promising applications due to its high availability,lower costs and existence of many glycerol-utilizing microorganisms....

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Main Author: Suhaimi, Sheril Norliana
Format: Thesis
Language:English
Published: 2013
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Online Access:http://psasir.upm.edu.my/id/eprint/48694/1/FBSB%202013%2043R.pdf
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spelling my-upm-ir.486942016-10-14T08:42:04Z Optimization of glycerol fermentation conditions for bioethanol production by locally isolated Escherichia coli SS1 2013-10 Suhaimi, Sheril Norliana The increase of biodiesel production worldwide lead to the increase of glycerol present in environment. Bioconverting glycerol into various valuable products is one of glycerol’s promising applications due to its high availability,lower costs and existence of many glycerol-utilizing microorganisms. Bioethanol is one of the interest products generated and its production is depending on the types of microorganism used and fermentation conditions. The effective screening procedure is needed to screen and isolate broad ranges of bacteria from environment. This study aims to isolate a suitable glycerol-fermenting microorganism and able to produce high ethanol production under optimized condition using crude glycerol as a substrate. The screening method was modified based on enzymatic oxidation of ethanol which is correlated to reduction of 2,6-dichlorophenol-indophenol dye. The formation of decolourized zone was apparent using modified assay containing 5 ml/L of 0.05 M 2,6-dichlorophenol-indophenol, 10 ml of reaction mixture and 500 μl/L of enzyme, respectively. In this study, several isolates were obtained showing capability in producing ethanol. The isolate namely E.coli SS1 was obtained after several screening processes. The fermentation of glycerol was carried out in batch fermentation using 120 mL serum bottle, incubated in 37°C with 120 rpm of agitation under anaerobic condition. The maximum ethanol production was achieved at 96 hours with 9.23 g/L, which is corresponding to the yield of 1 mol ethanol per mol glycerol and the productivity of 0.01 mol/molh-1. This isolate also showed a higher affinity to glycerol than glucose for bioethanol production. Response surface methodology (RSM) employed to obtain optimized condition. Six parameters were subjected to two level factorial design which were initial pH, substrate concentration, sodium chloride content, trace element solution, incubation temperature and the concentration of yeast extract and tryptone. The results showed that only 4 parameters were identified as significant factors, i.e initial pH, substrate concentration, sodium chloride content and the mixture of yeast extract and tryptone. The effect of the significant variables was subsequently evaluated in Central Composite Design (CCD) to determine the optimum value for each variable. The optimized conditions obtained were at initial pH of 7.61, substrate concentration of 34.5 g/L, the mixture of yeast extract and tryptone at 6.42 g/L whereas salt content was identified as a non-significant parameter, with predicted maximum ethanol production of 17.05 g/L (1.0 mol/mol). Avalidation run was performed using crude glycerol based on optimized conditions and maximum ethanol obtained was 15.72 ± 0.26 g/L with the yield of 1.0 mol/mol and productivity at 0.01 mol/mol.h-1 which was comparable to the predicted ethanol concentration. The isolated E.coli SS1 is a potential ethanol producer from glycerol, where the increased ethanol production is observed under optimized fermentation conditions. The crude glycerol also is feasible to be fermented using this isolate for comparable ethanol production using pure glycerol. Glycerin Escherichia coli 2013-10 Thesis http://psasir.upm.edu.my/id/eprint/48694/ http://psasir.upm.edu.my/id/eprint/48694/1/FBSB%202013%2043R.pdf application/pdf en public masters Universiti Putra Malaysia Glycerin Escherichia coli
institution Universiti Putra Malaysia
collection PSAS Institutional Repository
language English
topic Glycerin
Escherichia coli

spellingShingle Glycerin
Escherichia coli

Suhaimi, Sheril Norliana
Optimization of glycerol fermentation conditions for bioethanol production by locally isolated Escherichia coli SS1
description The increase of biodiesel production worldwide lead to the increase of glycerol present in environment. Bioconverting glycerol into various valuable products is one of glycerol’s promising applications due to its high availability,lower costs and existence of many glycerol-utilizing microorganisms. Bioethanol is one of the interest products generated and its production is depending on the types of microorganism used and fermentation conditions. The effective screening procedure is needed to screen and isolate broad ranges of bacteria from environment. This study aims to isolate a suitable glycerol-fermenting microorganism and able to produce high ethanol production under optimized condition using crude glycerol as a substrate. The screening method was modified based on enzymatic oxidation of ethanol which is correlated to reduction of 2,6-dichlorophenol-indophenol dye. The formation of decolourized zone was apparent using modified assay containing 5 ml/L of 0.05 M 2,6-dichlorophenol-indophenol, 10 ml of reaction mixture and 500 μl/L of enzyme, respectively. In this study, several isolates were obtained showing capability in producing ethanol. The isolate namely E.coli SS1 was obtained after several screening processes. The fermentation of glycerol was carried out in batch fermentation using 120 mL serum bottle, incubated in 37°C with 120 rpm of agitation under anaerobic condition. The maximum ethanol production was achieved at 96 hours with 9.23 g/L, which is corresponding to the yield of 1 mol ethanol per mol glycerol and the productivity of 0.01 mol/molh-1. This isolate also showed a higher affinity to glycerol than glucose for bioethanol production. Response surface methodology (RSM) employed to obtain optimized condition. Six parameters were subjected to two level factorial design which were initial pH, substrate concentration, sodium chloride content, trace element solution, incubation temperature and the concentration of yeast extract and tryptone. The results showed that only 4 parameters were identified as significant factors, i.e initial pH, substrate concentration, sodium chloride content and the mixture of yeast extract and tryptone. The effect of the significant variables was subsequently evaluated in Central Composite Design (CCD) to determine the optimum value for each variable. The optimized conditions obtained were at initial pH of 7.61, substrate concentration of 34.5 g/L, the mixture of yeast extract and tryptone at 6.42 g/L whereas salt content was identified as a non-significant parameter, with predicted maximum ethanol production of 17.05 g/L (1.0 mol/mol). Avalidation run was performed using crude glycerol based on optimized conditions and maximum ethanol obtained was 15.72 ± 0.26 g/L with the yield of 1.0 mol/mol and productivity at 0.01 mol/mol.h-1 which was comparable to the predicted ethanol concentration. The isolated E.coli SS1 is a potential ethanol producer from glycerol, where the increased ethanol production is observed under optimized fermentation conditions. The crude glycerol also is feasible to be fermented using this isolate for comparable ethanol production using pure glycerol.
format Thesis
qualification_level Master's degree
author Suhaimi, Sheril Norliana
author_facet Suhaimi, Sheril Norliana
author_sort Suhaimi, Sheril Norliana
title Optimization of glycerol fermentation conditions for bioethanol production by locally isolated Escherichia coli SS1
title_short Optimization of glycerol fermentation conditions for bioethanol production by locally isolated Escherichia coli SS1
title_full Optimization of glycerol fermentation conditions for bioethanol production by locally isolated Escherichia coli SS1
title_fullStr Optimization of glycerol fermentation conditions for bioethanol production by locally isolated Escherichia coli SS1
title_full_unstemmed Optimization of glycerol fermentation conditions for bioethanol production by locally isolated Escherichia coli SS1
title_sort optimization of glycerol fermentation conditions for bioethanol production by locally isolated escherichia coli ss1
granting_institution Universiti Putra Malaysia
publishDate 2013
url http://psasir.upm.edu.my/id/eprint/48694/1/FBSB%202013%2043R.pdf
_version_ 1747811993572605952