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首页> 外文期刊>Applied Microbiology and Biotechnology >Exploiting mixtures of H-2, CO2, and O-2 for improved production of methacrylate precursor 2-hydroxyisobutyric acid by engineered Cupriavidus necator strains
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Exploiting mixtures of H-2, CO2, and O-2 for improved production of methacrylate precursor 2-hydroxyisobutyric acid by engineered Cupriavidus necator strains

机译：利用H-2，CO2和O-2的混合物改善工程化Cupriavidus necator菌株生产甲基丙烯酸酯前体2-羟基异丁酸的能力

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Current manufacturing of most bulk chemicals through petrochemical routes considerably contributes to common concerns over the depletion of fossil carbon sources and greenhouse gas emissions. Sustainable future production of commodities thus requires the shift to renewable feedstocks in combination with established or newly developed synthesis routes. In this study, the potential of Cupriavidus necator H16 for autotrophic synthesis of the building block chemical 2-hydroxyisobutyric acid (2-HIBA) is evaluated. A novel biosynthetic pathway was implemented by heterologous expression of the 2-hydroxyisobutyryl-coenzyme A (2-HIB-CoA) mutase from Aquincola tertiaricarbonis L108, relying on a main intermediate of strain H16's C4 overflow metabolism, 3-hydroxybutyryl-CoA. The intention was to direct the latter to 2-HIBA instead or in addition to poly-3-hydroxybutyrate (PHB). Autotrophic growth and 2-HIBA (respectively, PHB) synthesis of wild-type and PHB-negative mutant strains were investigated producing maximum 2-HIBA titers of 3.2 g L-1 and maximum specific 2-HIBA synthesis rates (q (2-HIBA)) of about 16 and 175 mu mol g(-1) h(-1), respectively. The obtained specific productivity was the highest reported to date for mutase-dependent 2-HIBA synthesis from heterotrophic and autotrophic substrates. Furthermore, expression of a G protein chaperone (MeaH) in addition to the 2-HIB-CoA mutase subunits yielded improved productivity. Analyzing the inhibition of growth and product synthesis due to substrate availability and product accumulation revealed a strong influence of 2-HIBA, when cells were cultivated at high titers. Nevertheless, the presented results imply that at the time the autotrophic synthesis route is superior to thus far established heterotrophic routes for production of 2-HIBA with C. necator.
机译：当前通过石化途径生产大多数散装化学品的行为，极大地引起了人们对化石碳源消耗和温室气体排放的普遍关注。因此，商品的可持续未来生产需要结合已建立或新开发的合成路线，转向可再生原料。在这项研究中，评估了铜绿菌（Cupriavidus necator）H16在自养性合成基础化学2-羟基异丁酸（2-HIBA）方面的潜力。依赖于菌株H16的C4溢流代谢的主要中间体3-羟基丁酰辅酶A，通过异源表达来自Aquincola tertiaricarbonis L108的2-羟基异丁酰辅酶A（2-HIB-CoA）突变酶，实现了一种新的生物合成途径。目的是代替或除聚-3-羟基丁酸酯（PHB）外将后者导向2-HIBA。研究了野生型和PHB阴性突变菌株的自养生长和2-HIBA（分别为PHB）合成，产生的最大2-HIBA效价为3.2 g L-1，最大2-（HIBA）合成率最高（q（2-HIBA ））分别约为16和175μmol g（-1）h（-1）。迄今为止，从异养和自养底物合成的依赖突变酶的2-HIBA合成所获得的比生产率是最高的。此外，除2-HIB-CoA突变酶亚基外，G蛋白伴侣（MeaH）的表达还提高了生产率。分析由于底物可用性和产物积累而引起的生长和产物合成的抑制作用，揭示了当以高滴度培养细胞时2-HIBA的强烈影响。然而，提出的结果暗示，在那时自养合成路线优于迄今建立的用C.necator生产2-HIBA的异养路线。

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