Citation
Zakaria, Muhammad Azman
(2017)
Role of yqiG pseudogene in Escherichia coli towards biohydrogen production.
Masters thesis, Universiti Putra Malaysia.
Abstract
Biohydrogen gas has a great potential as alternative energy and it is considered as
environmental friendly energy source. Extensive studies have been carried out on the
production of biohydrogen from Escherichia coli (E. coli). E. coli was used due to its
accessibility of full genome and well characterized proteins. Interestingly, there are
some of pseudogenes present in E. coli genome. The pseudogenes were considered as
non-functional genes as result of genetic variability. In this study, we found that yqiG is
one of the pseudogenes in E. coli that is essential for biohydrogen production. The
deletion of yqiG has reduced the biohydrogen productivity in comparison to the E.
coli wild type as control. The objective of this study is to investigate the role of yqiG
pseudogene of E. coli during hydrogen production. The yqiG single gene deletion strain
and ASKA strains were used throughout the study. The yqiG mutant strain has
significantly interrupted biohydrogen productivity and suppressed glucose consumption
up to 50% compared to wild type, respectively. In addition, overexpressed yqiG protein
in the wild type via pCA24N-YqiG had increased the hydrogen productivity to 56
μmol H2/mg protein from glucose compared to control which only 38 μmol H2/mg
protein. Similar pattern was observed from formate fermentation whereas wild type
producing yqiG protein dominated the biohydrogen productivity about 141.8 μmol
H2/mg protein. Higher expression of formate hydrogen lyase components (fdhF, hycE
and fhlA) were observed in mutant yqiG strain. In contrast, the yqiG mutation causes
down regulation of pflB and pykF of glycolysis as well as formic acid accumulation in
the fermentation broth compared to the wild type strain. The pflB and pykF encode
pyruvate kinase and pyruvate formate lyase, respectively. We conclude that the yqiG
protein is important in the conversion of phosphoenolpyruvate to pyruvate and
utilization of pyruvate to formate for hydrogen production. Thus, yqiG pseudogene
makes an important contribution during hydrogen metabolism in E.coli.
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