Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae

Bioconversion of crude glycerol to biohydrogen is promising because the cost for capital investment and operation is cheaper, and it can help in reducing waste thus making it a clean process. However, not many microbes can metabolise glycerol efficiently under anaerobic or oxygen-limited conditions....

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Main Author:	Haron, Roslindawati
Format:	Thesis
Language:	English
Published:	2020
Subjects:	TP Chemical technology
Online Access:	http://eprints.utm.my/102644/1/RoslindawatiHaronPSChE2020.pdf.pdf
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spelling	my-utm-ep.1026442023-09-13T02:13:18Z Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae 2020 Haron, Roslindawati TP Chemical technology Bioconversion of crude glycerol to biohydrogen is promising because the cost for capital investment and operation is cheaper, and it can help in reducing waste thus making it a clean process. However, not many microbes can metabolise glycerol efficiently under anaerobic or oxygen-limited conditions. This makes the production and yield of hydrogen produced to be low. Therefore, this study sought to isolate new indigenous bacteria that can consume crude glycerol (85%) and convert it into biohydrogen without the need for pretreatment or acclimatization. Dark fermentation approach was employed since this approach offers much advantages in terms of substrates and microbe producers used. The aim of study was achieved through isolation and screening for potential hydrogen producers followed by identification using 16S rRNA technique. Next, optimisation of medium composition and optimisation of operating parameters to obtain the optimum production and yield of hydrogen and ethanol including glycerol uptake. The study was finalised by a kinetic study for growth and substrate utilisation by kinetic model and potential hydrogen production by modified Gompertz model. The best hydrogen producer had been successfully isolated, screened and identified as Klebsiella pneumoniae strain HS11286. The 2-level fractional factorial design (28–3) for medium composition optimisation and response surface methodology study employing Box-Behnken design for operating parameters were employed to evaluate the interactive effects of several respective factors. The fermentation was conducted in 2 L reactor with a working volume of 1.8 L. Medium optimisation study gave the composition of 30 g/L glycerol, 3.5 g/L K2HPO4, 3.98g/L KH2PO4, 2.69 g/L (NH4)2SO4, 0.03 g/L CaCl2.2H20, 0.054 g/L FESO4.7H2O, 3.0 g/L yeast extract, and 0.54 g/L MgSO4.7H2O as the optimised medium composition. This optimised composition yielded 588.68±0.04 mmol H2/mol glycerolconsumed, 9345±63.64 mL of hydrogen, 97±1.41% glycerol uptake and 0.024±0.001 mmol ethanol/mol glycerolconsumed. Meanwhile, the optimum operating condition was found best at pH 6.0, temperature 32.5°C and 25% headspace with 82% desirability. This yielded 601.07±10.69 mmol H2/mol glycerolconsumed of hydrogen yield, 9935±176 mL of hydrogen, 97±1.4% of glycerol uptake and 0.045±0.002 mmol ethanol/mol glycerolconsumed. Finally, kinetic parameters for specific growth rate (μ) was at 0.106 h−1, glycerol consumption rate (Qgly) at 1.572 g/L/h, and yield coefficient Yp/x, Yp/s and Yx/s at 30758.51 mL/g cell, 479.26 mL/g substrate and 0.016 g cell/g substrate, respectively. Meanwhile, the modified Gompertz model gave a prediction of 10155 mL of hydrogen at 620 mL/h. In conclusion, Klebsiella pneumonia strain HS11286 has the potential to produce almost 10 litre of hydrogen in a short period (less than 48 h) without the need to pretreat the glycerol or to acclimatize the bacteria in crude glycerol. 2020 Thesis http://eprints.utm.my/102644/ http://eprints.utm.my/102644/1/RoslindawatiHaronPSChE2020.pdf.pdf application/pdf en public http://dms.library.utm.my:8080/vital/access/manager/Repository/vital:149288 phd doctoral Universiti Teknologi Malaysia Faculty of Engineering - School of Chemical & Energy Engineering
institution	Universiti Teknologi Malaysia
collection	UTM Institutional Repository
language	English
topic	TP Chemical technology
spellingShingle	TP Chemical technology Haron, Roslindawati Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae
description	Bioconversion of crude glycerol to biohydrogen is promising because the cost for capital investment and operation is cheaper, and it can help in reducing waste thus making it a clean process. However, not many microbes can metabolise glycerol efficiently under anaerobic or oxygen-limited conditions. This makes the production and yield of hydrogen produced to be low. Therefore, this study sought to isolate new indigenous bacteria that can consume crude glycerol (85%) and convert it into biohydrogen without the need for pretreatment or acclimatization. Dark fermentation approach was employed since this approach offers much advantages in terms of substrates and microbe producers used. The aim of study was achieved through isolation and screening for potential hydrogen producers followed by identification using 16S rRNA technique. Next, optimisation of medium composition and optimisation of operating parameters to obtain the optimum production and yield of hydrogen and ethanol including glycerol uptake. The study was finalised by a kinetic study for growth and substrate utilisation by kinetic model and potential hydrogen production by modified Gompertz model. The best hydrogen producer had been successfully isolated, screened and identified as Klebsiella pneumoniae strain HS11286. The 2-level fractional factorial design (28–3) for medium composition optimisation and response surface methodology study employing Box-Behnken design for operating parameters were employed to evaluate the interactive effects of several respective factors. The fermentation was conducted in 2 L reactor with a working volume of 1.8 L. Medium optimisation study gave the composition of 30 g/L glycerol, 3.5 g/L K2HPO4, 3.98g/L KH2PO4, 2.69 g/L (NH4)2SO4, 0.03 g/L CaCl2.2H20, 0.054 g/L FESO4.7H2O, 3.0 g/L yeast extract, and 0.54 g/L MgSO4.7H2O as the optimised medium composition. This optimised composition yielded 588.68±0.04 mmol H2/mol glycerolconsumed, 9345±63.64 mL of hydrogen, 97±1.41% glycerol uptake and 0.024±0.001 mmol ethanol/mol glycerolconsumed. Meanwhile, the optimum operating condition was found best at pH 6.0, temperature 32.5°C and 25% headspace with 82% desirability. This yielded 601.07±10.69 mmol H2/mol glycerolconsumed of hydrogen yield, 9935±176 mL of hydrogen, 97±1.4% of glycerol uptake and 0.045±0.002 mmol ethanol/mol glycerolconsumed. Finally, kinetic parameters for specific growth rate (μ) was at 0.106 h−1, glycerol consumption rate (Qgly) at 1.572 g/L/h, and yield coefficient Yp/x, Yp/s and Yx/s at 30758.51 mL/g cell, 479.26 mL/g substrate and 0.016 g cell/g substrate, respectively. Meanwhile, the modified Gompertz model gave a prediction of 10155 mL of hydrogen at 620 mL/h. In conclusion, Klebsiella pneumonia strain HS11286 has the potential to produce almost 10 litre of hydrogen in a short period (less than 48 h) without the need to pretreat the glycerol or to acclimatize the bacteria in crude glycerol.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Haron, Roslindawati
author_facet	Haron, Roslindawati
author_sort	Haron, Roslindawati
title	Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae
title_short	Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae
title_full	Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae
title_fullStr	Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae
title_full_unstemmed	Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae
title_sort	optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae
granting_institution	Universiti Teknologi Malaysia
granting_department	Faculty of Engineering - School of Chemical & Energy Engineering
publishDate	2020
url	http://eprints.utm.my/102644/1/RoslindawatiHaronPSChE2020.pdf.pdf
_version_	1783729200496115712

Optimization of media and process parameters for bioconversion of crude glycerol to hydrogen by locally isolated klebsiella pneumoniae

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