Enhanced production of d-β-hydroxyisobutyric acid through strain improvement

Kim, Chul Ho; Hong, Won Kyoung; Lee, In Young; Choi, Eui Sung; Rhee, Shin Hyung

doi:10.1016/s0168-1656(99)00003-6

Cited by 4 publications

(2 citation statements)

References 3 publications

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“…strain VLB120 to grow on isobutanol, isobutyraldehyde and isobutyric acid (Table 2 ). The existence of an isobutyric acid degradation pathway was already described for P. putida , Candida rugosa , Yarrowia lipolytica and Desulfococcus multivorans and used in mutant strains of the first three species for the synthesis of 3-hydroxyisobutyric acid using isobutyric acid as precursor [ 36 , 42 - 45 ].…”

Section: Discussionmentioning

confidence: 99%

Metabolic engineering of Pseudomonas sp. strain VLB120 as platform biocatalyst for the production of isobutyric acid and other secondary metabolites

et al. 2014

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BackgroundOver the recent years the production of Ehrlich pathway derived chemicals was shown in a variety of hosts such as Escherichia coli, Corynebacterium glutamicum, and yeast. Exemplarily the production of isobutyric acid was demonstrated in Escherichia coli with remarkable titers and yields. However, these examples suffer from byproduct formation due to the fermentative growth mode of the respective organism. We aim at establishing a new aerobic, chassis for the synthesis of isobutyric acid and other interesting metabolites using Pseudomonas sp. strain VLB120, an obligate aerobe organism, as host strain.ResultsThe overexpression of kivd, coding for a 2-ketoacid decarboxylase from Lactococcus lactis in Ps. sp. strain VLB120 enabled for the production of isobutyric acid and isobutanol via the valine synthesis route (Ehrlich pathway). This indicates the existence of chromosomally encoded alcohol and aldehyde dehydrogenases catalyzing the reduction and oxidation of isobutyraldehyde. In addition we showed that the strain possesses a complete pathway for isobutyric acid metabolization, channeling the compound via isobutyryl-CoA into valine degradation. Three key issues were addressed to allow and optimize isobutyric acid synthesis: i) minimizing isobutyric acid degradation by host intrinsic enzymes, ii) construction of suitable expression systems and iii) streamlining of central carbon metabolism finally leading to production of up to 26.8 ± 1.5 mM isobutyric acid with a carbon yield of 0.12 ± 0.01 g gglc-1.ConclusionThe combination of an increased flux towards isobutyric acid using a tailor-made expression system and the prevention of precursor and product degradation allowed efficient production of isobutyric acid in Ps. sp. strain VLB120. This will be the basis for the development of a continuous reaction process for this bulk chemicals.

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Section: Discussionmentioning

confidence: 99%

Metabolic engineering of Pseudomonas sp. strain VLB120 as platform biocatalyst for the production of isobutyric acid and other secondary metabolites

et al. 2014

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“…The biocatalyst allowing higher yields of the product to be obtained was selected from among the mutants of C. rugosa IFO 0750 (mutations induced by UV/chemical treatments), which were not able to assimilate propionic acid. A stable mutant of C. rugosa MME 1259 transformed isobutyric acid to ( R )-β-hydroxyisobutyric acid in a concentration of 150 g/L and with the molar conversion yield of over 80% [84]. ( R )-β-Hydroxyisobutyric acid is a building block for the synthesis of captopril, which is known as a blood pressure-lowering agent [83] (Figure 15).…”

Section: Production Of Optically Active Compoundsmentioning

confidence: 99%

Biotransformations Utilizing β-Oxidation Cycle Reactions in the Synthesis of Natural Compounds and Medicines

Œwizdor

Panek

Milecka-Tronina

et al. 2012

IJMS

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β-Oxidation cycle reactions, which are key stages in the metabolism of fatty acids in eucaryotic cells and in processes with a significant role in the degradation of acids used by microbes as a carbon source, have also found application in biotransformations. One of the major advantages of biotransformations based on the β-oxidation cycle is the possibility to transform a substrate in a series of reactions catalyzed by a number of enzymes. It allows the use of sterols as a substrate base in the production of natural steroid compounds and their analogues. This route also leads to biologically active compounds of therapeutic significance. Transformations of natural substrates via β-oxidation are the core part of the synthetic routes of natural flavors used as food additives. Stereoselectivity of the enzymes catalyzing the stages of dehydrogenation and addition of a water molecule to the double bond also finds application in the synthesis of chiral biologically active compounds, including medicines. Recent advances in genetic, metabolic engineering, methods for the enhancement of bioprocess productivity and the selectivity of target reactions are also described.

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Multistep Synthesis of (S)‐3‐Hydroxyisobutyric Acid from Glucose usingPseudomonas taiwanensisVLB120 B83 T7 Catalytic Biofilms

2015

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Thec hiral building block (S)-3-hydroxyisobutyric acid [(S)-3-HIBA] was produced either by conversion of isobutyric acid or directly from glucose utilizing cells of Pseudomonas taiwanensis VLB120 B83 T7 as catalysts.T his strain carriesapoint mutation in the gene encoding 3-hydroxyisobutyrate dehydrogenase (mmsB), leading to its inactivation.T he maximal specific activity in resting-cell biotransformations using isobutyric acid as substrate was 4.9 AE 0.4 Ug cdw À1 .O verexpression of the 2-ketoisovalerate pathwayg enes alsS,i lvC, and ilvD,a nd the introduction of kivd encoding 2-ketoacid decarboxylase resulted in the efficient fermentatives ynthesiso f( S )-3-HIBAd irectly from glucose.U pt o2 2mM( 2.3 g L À1 ) (S)-3-HIBAwere produceda t3 .7 AE 0.3 Ug cdw À1 in repeated batch experiments without observable product degradation. Utilizing ab iofilm reactor it was possible to continuously produce up to.O verall, the conversion of isobutyric acid to (S)-3-HIBAw as found to be the rate-limiting step,l eading to the accumulation of am ixture of (S)-3-hydroxyisobutyric acid andi sobutyric acid. This study demonstratesf or the first time the production of (S)-3-HIBAf rom renewable carbon in shake flasks and biofilm reactors and sets the stage for furthero ptimizations towards the efficient production of 3-HIBAa nd its derivatives in continuous fermentations.

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Enhanced production of d-β-hydroxyisobutyric acid through strain improvement

Cited by 4 publications

References 3 publications

Metabolic engineering of Pseudomonas sp. strain VLB120 as platform biocatalyst for the production of isobutyric acid and other secondary metabolites

Metabolic engineering of Pseudomonas sp. strain VLB120 as platform biocatalyst for the production of isobutyric acid and other secondary metabolites

Biotransformations Utilizing β-Oxidation Cycle Reactions in the Synthesis of Natural Compounds and Medicines

Multistep Synthesis of (S)‐3‐Hydroxyisobutyric Acid from Glucose usingPseudomonas taiwanensisVLB120 B83 T7 Catalytic Biofilms

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