Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation

Effect of growth conditions (temperature, agitation rate and working volume) on production of enhanced green fluorescent protein (EGFP) from Escherichia coli (E.coli) fermentation was studied in this research. An improved growth conditions are needed to maximize the functional EGFP production. Prepa...

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Main Author:	Malini, Subramaniam
Format:	Undergraduates Project Papers
Language:	English
Published:	2014
Subjects:	QP Physiology
Online Access:	http://umpir.ump.edu.my/id/eprint/8851/ http://umpir.ump.edu.my/id/eprint/8851/ http://umpir.ump.edu.my/id/eprint/8851/1/CD8546.pdf

id	ump-8851
recordtype	eprints
spelling	ump-88512015-10-26T06:25:10Z http://umpir.ump.edu.my/id/eprint/8851/ Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation Malini, Subramaniam QP Physiology Effect of growth conditions (temperature, agitation rate and working volume) on production of enhanced green fluorescent protein (EGFP) from Escherichia coli (E.coli) fermentation was studied in this research. An improved growth conditions are needed to maximize the functional EGFP production. Preparation of cell culture was done by transferring E. coli strain BL21 (DE3) carrying plasmid pRSETEGFP to agar plate by streaking method and incubated at 30°C for 18 hr. Inoculum was prepared from a single colony of E.coli from agar plate. Fermentation of batch cultures were carried out in Erlenmeyer flasks inoculated with inocula [5% (v/v)]. The process variables include temperature, agitation rate and working volume were varied throughout the batch fermentation by using one factor at a time method. During the cultivation process, samples were taken from Erlenmeyer flask to measure cell biomass and EGFP concentrations. Cell biomass concentration was determined based on the culture absorbance using spectrophotometer and EGFP concentration was determined using gel-based imaging method. The results obtained shows that EGFP production by E.coli decreases as the working volumes was increased from 20 to 50%. However, when the agitation rate was increased from 100 to 250 rpm, it subsequently increases the yield of EGFP and cell concentration. Suitable temperature (30°C) enables E.coli to grow well and produce higher amount of functional EGFP. Consequently, functional EGFP production decreased at low temperature (<30°C). At higher temperature (> 30°C) also resulted in a decrease of EGFP and biomass production due to protein aggregation into inclusion bodies. In conclusion, the yield of functional EGFP was highest in shake flask fermentation under condition, working volume of 20%, agitation rate of 200 rpm and temperature of 30°C with 0.04, 0.061 and 0.06 g/L respectively 2014-01 Undergraduates Project Papers NonPeerReviewed application/pdf en http://umpir.ump.edu.my/id/eprint/8851/1/CD8546.pdf Malini, Subramaniam (2014) Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation. Faculty of Chemical & Natural Resources Engineering, Universiti Malaysia Pahang. http://iportal.ump.edu.my/lib/item?id=chamo:84870&theme=UMP2
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Malaysia Pahang
building	UMP Institutional Repository
collection	Online Access
language	English
topic	QP Physiology
spellingShingle	QP Physiology Malini, Subramaniam Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation
description	Effect of growth conditions (temperature, agitation rate and working volume) on production of enhanced green fluorescent protein (EGFP) from Escherichia coli (E.coli) fermentation was studied in this research. An improved growth conditions are needed to maximize the functional EGFP production. Preparation of cell culture was done by transferring E. coli strain BL21 (DE3) carrying plasmid pRSETEGFP to agar plate by streaking method and incubated at 30°C for 18 hr. Inoculum was prepared from a single colony of E.coli from agar plate. Fermentation of batch cultures were carried out in Erlenmeyer flasks inoculated with inocula [5% (v/v)]. The process variables include temperature, agitation rate and working volume were varied throughout the batch fermentation by using one factor at a time method. During the cultivation process, samples were taken from Erlenmeyer flask to measure cell biomass and EGFP concentrations. Cell biomass concentration was determined based on the culture absorbance using spectrophotometer and EGFP concentration was determined using gel-based imaging method. The results obtained shows that EGFP production by E.coli decreases as the working volumes was increased from 20 to 50%. However, when the agitation rate was increased from 100 to 250 rpm, it subsequently increases the yield of EGFP and cell concentration. Suitable temperature (30°C) enables E.coli to grow well and produce higher amount of functional EGFP. Consequently, functional EGFP production decreased at low temperature (<30°C). At higher temperature (> 30°C) also resulted in a decrease of EGFP and biomass production due to protein aggregation into inclusion bodies. In conclusion, the yield of functional EGFP was highest in shake flask fermentation under condition, working volume of 20%, agitation rate of 200 rpm and temperature of 30°C with 0.04, 0.061 and 0.06 g/L respectively
format	Undergraduates Project Papers
author	Malini, Subramaniam
author_facet	Malini, Subramaniam
author_sort	Malini, Subramaniam
title	Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation
title_short	Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation
title_full	Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation
title_fullStr	Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation
title_full_unstemmed	Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation
title_sort	effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation
publishDate	2014
url	http://umpir.ump.edu.my/id/eprint/8851/ http://umpir.ump.edu.my/id/eprint/8851/ http://umpir.ump.edu.my/id/eprint/8851/1/CD8546.pdf
first_indexed	2023-09-18T22:06:52Z
last_indexed	2023-09-18T22:06:52Z
_version_	1777414766528036864

Effect of growth conditions on production of green fluorescent protein from escherichia coli fermentation

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