Inverted metabolic engineering of E. coli toproduce PHB and simplifying of lysis process, using gene E from phage PhiX۱۷۴

AuthorsRoohollah Nakhaei Sistani- Hadi Shirzad- Mojtaba Saadati
JournalJournal of Army University
Paper TypeFull Paper
Published At۲۰۱۰
Journal GradeScientific - research
Journal TypeTypographic
Journal CountryIran, Islamic Republic Of

Abstract

Background: The polyhydroxybutyrate biosynthetic genes of Ralstonia eutropha are organized in single
operon. There are many studies which show that this operon could be cloned in gram negative bacteria
like E. coli. As its original promoter could work efficiently in E. coli, there is no need to change it with host
ones. Granule extraction is one of the most important considerations of industrial production of PHB. Solvent
base or physical approaches increase the cost of production and compromise the integrity of PHB granules.
Therefore, E mediated lysis was used in this study to extract the granules.
Materials and Methods: In this study, the whole operon of phb from Ralstonia eutropha and E gene from
phage phiX174 were obtained by PCR technique, and cloned transferred to E. coli by separate plasmids. To
control the lysis process, the chemical inducible system was used.
Results: Bacterial cells which have both plasmids could produce high levels of PHB, and their PHB
content could be released into the medium, nearly perfectly, at the correct time by adding IPTG as a chemical
inducer.
Conclusion: This method could be used to produce and extract PHB with more cost effectiveness in
industrial scales.

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