optimization of auto-induction conditions for the heterologous expression of a maltogenic amylase in escherichia coli

Authors

zahra goudarzi

seyed abbas shojaosadati department of chemical engineering, tarbiat modares university, tehran

reza hassan sajedi

amir maghsoudi

abstract

background and objectives: auto-induction is usually employed to achieve high cell density and overproduction of proteins with a simple and low-cost operation. the efficiency of heterologous protein expression in escherichia coli is determined by different parameters. interactions between these parameters usually complicate the identification of those that contribute more to the improvement of protein expression. as optimal implementation of the auto-induction considerably relies on both the composition of the auto-induction medium and induction conditions, the present study focused on the optimization of related culture parameters through response surface methodology.   materials and methods: in the first step, the optimum culture temperature and auto-induction duration were determined with the aim of achieving the highest specific activity. then the culture composition was optimized through response surface methodology considering the concentration of carbon sources, glucose and lactose, as the variables for the simultaneous maximizing of the maltogenic amylase volumetric yield and specific activity.   results and conclusion: expression of recombinant maltogenic amylase under optimum conditions in the shake-flask cultures of escherichia coli harboring pet 28a increased by 1.7 folds in comparison with an un-optimized auto-induction culture. the maltogenic amylase specific activity and volumetric yield were found to be 34.93 u mg -1 and 390.78 u ml -1 at optimum conditions, respectively.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

Enhanced Expression of Recombinant Activin A in Escherichia coli by Optimization of Induction Parameters

Activin A is a member of the transforming growth factor β super family. Because of its extensive clinical usages, its recombinant production is beneficial. In this study, activin A was expressed in E. coli using the pET 21a expression vector. The optimization of the activin A production in E. coli was done by using the response surface methodology (RSM). At this stage, the effect of IPTG and la...

full text

the effect of taftan pozzolan on the compressive strength of concrete in the environmental conditions of oman sea (chabahar port)

cement is an essential ingredient in the concrete buildings. for production of cement considerable amount of fossil fuel and electrical energy is consumed. on the other hand for generating one tone of portland cement, nearly one ton of carbon dioxide is released. it shows that 7 percent of the total released carbon dioxide in the world relates to the cement industry. considering ecological issu...

study of cohesive devices in the textbook of english for the students of apsychology by rastegarpour

this study investigates the cohesive devices used in the textbook of english for the students of psychology. the research questions and hypotheses in the present study are based on what frequency and distribution of grammatical and lexical cohesive devices are. then, to answer the questions all grammatical and lexical cohesive devices in reading comprehension passages from 6 units of 21units th...

Optimization of Cloning Conditions for high-level Production of Recombinant Mouse Interleukin-2 in Escherichia coli

Backgrounds and objectives: Interleukin 2 (IL-2) secreted by activated CD4+ T cells  has been known as a major mediator in both  adaptive and native immune system  due to a board range of effects on  different cells in the immunity system (1-6). Methods: cDNA synthesis was performed using gene- specific primers designed by Gene Runner software after RNA extraction of mouse splenocytes. PCR pro...

full text

The Expression of Human Granulocyte Macrophage Colony Stimulating Factor by Heat-Induction in Escherichia coli

A self-regulated high-copy number plasmid containing chloramphenicol resistant gene, for the production of recombinant proteins under the regulation of bacteriophage ?pL promoter, was constructed. The designed 5024 base pair expression plasmid contained a heat sensitive repressor cI857 coding gene to regulate the function of ?pL promoter under heat shock induction. Using the constructed vector,...

full text

My Resources

Save resource for easier access later


Journal title:
applied food biotechnology

جلد ۳، شماره ۲، صفحات ۱۰۵-۱۱۳

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023