Abstract

Abstract Background The synthesis of 2-hydroxyisobutyric acid (2-HIB), a promising building block for, e.g., Plexiglas® production, is described as an example for a clean and sustainable bioproduction. Methods A derivative strain of Cupriavidus necator H16, impaired in the poly-ß-hydroxybutyrate synthesis pathway and equipped with xenogenic 2-hydroxyisobutyryl-coenzyme A mutase from Aquincola tertiaricarbonis L108, was applied. Batch cultivation was performed in the presence of vitamin B12 by supplying a gas mixture comprising hydrogen, oxygen, and carbon dioxide. Results Exploiting the chemo-litho-autotrophic potential of this so-called knallgas bacterium, 2-HIB was synthesized and excreted into the cultivation broth under aerobic conditions when inorganic nitrogen-limited conditions allowed an overflow metabolism of carbon metabolites. 2-HIB synthesis proceeded at a rate of 8.58 mg/[(g bacterial dry mass)·h]. Approximately 400 mg/L in total was obtained. The results were subsequently compared to calculated model data to evaluate the efficiency of the conversion of the substrates into the product. To achieve overall yield data regarding the substrate conversion, the model describes an integral process which includes both 2-HIB synthesis and biomass formation. Conclusions This study has confirmed the feasibility of the microbial synthesis of the bulk chemical 2-HIB from hydrogen and carbon dioxide by exploiting the chemo-litho-autotrophic metabolism of C. necator H16 PHB−4, additionally expressing the foreign 2-HIB-coenzyme A mutase. The product synthesis was satisfying as a proof of principle but does not yet approach the maximum value as derived from the model data. Furthermore, the biosynthesis potential of an optimized process is discussed in view of its technical application.

Highlights

  • The synthesis of 2-hydroxyisobutyric acid (2-HIB), a promising building block for, e.g., PlexiglasW production, is described as an example for a clean and sustainable bioproduction

  • We recently discovered a novel enzyme, the 2-HIBcoenzyme A mutase, which proves to be an ideal catalyst for the production of 2-HIB, especially, given that 2-HIB synthesis with this enzyme only requires a one-step isomerization of metabolites that are essential for the metabolism of a wide range of bacteria, i.e., 3-hydroxybutyryl-coenzyme A (3-HB-CoA) [25,41,42,43,44]

  • On-line analysis The gas concentrations were measured by specific sensors for hydrogen (0% to 100%), oxygen (0% to 100%), and carbon dioxide (0% to 50%) (BlueSens, Herten, Germany) and were continuously monitored

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Summary

Introduction

The synthesis of 2-hydroxyisobutyric acid (2-HIB), a promising building block for, e.g., PlexiglasW production, is described as an example for a clean and sustainable bioproduction. It is that fossil carbon sources will become limited in the future, but there is a growing pressure to renounce the exploitation of currently treated and prospected sites for environmental reasons. Accidents, such as the recent oil disaster in the Gulf of Mexico, motivate this tendency even more. The turnaround in thinking and acting has been already evident in recent times, mostly with respect to energy production, for which sustainable resources and clean techniques are increasingly implemented to substitute today's oil and coalbased production [1].

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