Publication Details |
Category | Text Publication |
Reference Category | Journals |
DOI | 10.1007/s00253-013-5064-x |
Document | Shareable Link |
Title (Primary) | Synthesis of the building block 2-hydroxyisobutyrate from fructose and butyrate by Cupriavidus necator H16 |
Author | Przybylski, D.; Rohwerder, T.; Harms, H.; Yaneva, N.; Müller, R.H. |
Source Titel | Applied Microbiology and Biotechnology |
Year | 2013 |
Department | UMB |
Volume | 97 |
Issue | 20 |
Page From | 8875 |
Page To | 8885 |
Language | englisch |
Keywords | 2-Hydroxyisobutyryl-CoA mutase; 2-Hydroxyisobutyric acid; Cupriavidus necator; H16 PHB−4; Fructose; Butyrate; Overflow metabolism |
UFZ wide themes | RU4; |
Abstract |
2-Hydroxyisobutyryl-coenzyme A mutase, originally discovered in the context of methyl tert-butyl ether degradation in Aquincola tertiaricarbonis L108, catalyzes the isomerization of 3-hydroxybutyryl-coenzyme A (3-HB-CoA) to 2-hydroxyisobutyryl-CoA. It thus constitutes the basis for a biotechnological route from practically any renewable carbon to 2-hydroxyisobutyrate (2-HIB) via the common metabolite 3-hydroxybutyrate. At first sight, recombinant Cupriavidus necator H16 expressing the mutase seems to be well suited for such a synthesis process, as a strong overflow metabolism via (R)-3-HB-CoA is easily induced in this bacterium possessing the poly-3-hydroxybutyrate metabolism. However, the recently established stereospecificity of the mutase, dominantly preferring the (S)-enantiomer of 3-HB-CoA, calls for a closer investigation of C. necator as potential 2-HIB production strain and raised the question about the strain’s potential to yield 2-HIB from substrates directly providing (S)-3-HB-CoA. We compared two mutase-expressing C. necator H16 strains for their capability to synthesize 2-HIB from fructose and butyrate, delivering either (R)- or (S)-3-HB-CoA. Our results indicate that due to the enantiospecificity of the mutase, fructose is a weaker substrate for 2-HIB synthesis than butyrate. Production rates achieved with the PHB-negative strain H16 PHB−4 on butyrate were higher than on fructose. Using the wild-type did not significantly improve the production rates as the latter showed a 34-fold and a 5-fold lower 2-HIB synthesis rate compared to H16 PHB−4 on fructose and butyrate, respectively. Moreover, both strains showed concomitant excretion of undesired side products, such as pyruvate and 3-hydroxybutyrate, significantly decreasing the 2-HIB yield. |
Persistent UFZ Identifier | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=14137 |
Przybylski, D., Rohwerder, T., Harms, H., Yaneva, N., Müller, R.H. (2013): Synthesis of the building block 2-hydroxyisobutyrate from fructose and butyrate by Cupriavidus necator H16 Appl. Microbiol. Biotechnol. 97 (20), 8875 - 8885 10.1007/s00253-013-5064-x |