Extended Abstract Journal of Genetics and DNA Research Volume 5:3, 2021

Open Access

Investigation of itaconate in Necator H16

Samuel Yemofio

University of Nottingham, UK, E-mail: [email protected]

Abstract

Recent challenges of pollution and climate change in our It is, therefore, necessary for the current generation to identify environment stems from the over-dependence on fossil fuel various sustainable and cleaner processes for chemical, fuel through the extraction, processing, and exploitation for and energy production. petrochemical-based products. This has caused severe havoc This study revealed that other genes can be involved in to the environment and its natural habitats, leading to deaths itaconate degradation and therefore further research to and displacements into unfavorable conditions. Researchers investigate the function of these genes is required. in the US Department of Energy (DoE) in 2004 identified In Australia, RLN species Pratylenchus thornei and P. itaconate, one of the twelve attractive platform chemicals, as neglectus are particularly important biotic constraints to wheat a potential chemical suitable for bio-based industrial products production. The most efficient and effective strategy for using biological routes. Previous research has also shown HPLC. that itaconate has the potential to replace petroleumbased Cupriavidus necator H16 (also known as Ralstonia eutropha products such as petrochemical-based acrylic and methacrylic H16) is a Gram-negative bacterium that belongs to the order acid; and detergents, surface active agents and , class . H16 has been biosynthesized plastics for industrial applications with bio- isolated from a near Goettingen, Germany, almost 60 based products. This can be achieved through biological or years ago (Schlegel et al. 1961a, b; Wilde 1962). Since then, chemical conversions and be subsequently converted into it has become the most-studied hydrogen-oxidising ‘Knallgas’ several high-value bio-based chemicals and materials from bacterium with best-characterised poly(3-hydroxybutyrate) biomass. Research also discovered that itaconate is naturally (PHB) metabolism. Cupriavidus necator H16 is a facultative produced by microorganisms such as Candida sp., Ustilago anaerobe, which can switch to anaerobic respiration by using madis and Aspergillus terreus although many microorganisms NO3 and NO2 as electron acceptors. It grows utilizing a have been genetically engineered for the biosynthesis of variety of organic substrates ranging from sugars, fatty acids, itaconate. It was used to estimate the concentration of amino acids and citric acid cycle intermediates. It is also itaconate consumed. The purpose of this research was to capable to fix directly a carbon dioxide (CO2) through the identify the genes involved in itaconate metabolism and CBB cycle using hydrogen as the energy source (Bowien and abolish its metabolism. To investigate itaconate metabolism Kusian 2002), making it a useful organism for studying on host organism Cupriavidus necator H16, the growth of mixotrophic and autotrophic metabolism. mutants was observed using itaconate as a sole carbon source. Single, double and triple knock-outs of ict genes involved in itaconate conversion to itaconyl-CoA (itaconate- CoA transferase activity) were generated. Growth and itaconate consumption assays were performed establishing that only H16_RS22140 gene is clearly involved in itaconate metabolism.

This work is partly presented at International Conference on Microbial Genetics and Molecular Microbiology, December 03-04, 2018

International Conference on Microbial Genetics and Molecular Microbiology 1 December 03-04, 2018