2-Hydroxyacyl-CoA lyase catalyzes acyloin condensation for one-carbon bioconversion

Chou, Alexander; Clomburg, James M.; Qian, Shuai; González, Ramón

doi:10.1038/s41589-019-0328-0

Cited by 65 publications

(91 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…During prolonged cultivation of strain DSM 45062 on 2-HIBA under selenium-limiting conditions, we observed a molar ratio of formic acid accumulation versus 2-HIBA consumption of only 0.6. This slightly understoichiometric accumulation might be resulting from partial assimilation of formic acid, possibly via reduction to formaldehyde and lyase-catalyzed acyloin condensation with formyl-CoA to glycolyl-CoA, as has been demonstrated for other 2-hydroxyacyl-CoA lyases (Chou et al, 2019). However, the novel lyase of strain DSM 45062…”

Section: Discussionmentioning

confidence: 81%

“…Likewise, suitable purification protocols have to be established, as our preliminary attempts to remove elution buffer and to concentrate the lyase after standard immobilized metal ion affinity chromatography resulted in precipitation of the heterologous protein. The thus far characterized 2-hydroxyacyl-CoA lyase from Rhodospirillales bacterium URHD0017 (Chou et al, 2019) shows a relatively low K m value with formyl-CoA (K m = 200 ± 50 µM) for the synthase reaction with shortchain aldehydes and ketones. However, the latter compounds are poor substrates which leads to low catalytic efficiency values for the production of 2-hydroxyacyl-CoAs.…”

Section: Discussionmentioning

confidence: 98%

“…Considering the abundance of the enzyme in 2-HIBA-grown cells (Table 1), a specific activity of about 250 nmol min −1 mg −1 (corresponding to a k cat of 0.26 s −1 ) can be deduced, indicating that enzyme preparation and assay protocols still need to be optimized. In addition, a significant overexpression in E. coli could not be established, likely due to insufficient soluble expression as previously observed for human HACL1 and other 2-hydroxyacyl-CoA lyases from prokaryotic origin (Chou et al, 2019;Burgener et al, 2020). In order to purify the enzyme from strain DSM 45062 for kinetic and structural characterization, production of the recombinant protein has to be improved, for example, by employing an actinobacterial expression system such as M. smegmatis (Bashiri and Baker, 2015).…”

Section: Discussionmentioning

confidence: 99%

“…The human peroxisomal 2hydroxyphytanoyl-CoA lyase HACL1 (NP_036392.2), for example, catalyzes the decomposition of 2-hydroxyphytanoyl-CoA (2-hydroxy-3,7,11,15-tetramethylhexadecanoyl-CoA) to the aldehyde pristanal (2,6,10,14-tetramethylpentadecanal) and formyl-CoA (Foulon et al, 1999(Foulon et al, , 2005. In addition, it has recently been demonstrated that HACL1 and related prokaryotic TPP-dependent enzymes can even catalyze the reversible acyloin condensation of formyl-CoA with shortand medium-chain carbonyl compounds (Chou et al, 2019;Burgener et al, 2020). Although these 2-hydroxyacyl-CoA lyases and the putative lyase from DSM 45062 are only distantly related (<30% sequence identity at about 90% query coverage), this provisional, only sequence-based functional assignment of WP_018331913.1 let us speculate about a mutase-independent pathway for the degradation of MPD and 2-HIBA proceeding via the decomposition of 2-hydroxyisobutyryl-CoA to acetone and formyl-CoA ( Figure 1A).…”

Section: Organization Of the Lyase-hcl Gene Cluster In A Chiangmaienmentioning

confidence: 99%

See 3 more Smart Citations

Actinobacterial Degradation of 2-Hydroxyisobutyric Acid Proceeds via Acetone and Formyl-CoA by Employing a Thiamine-Dependent Lyase Reaction

et al. 2020

View full text Add to dashboard Cite

The tertiary branched short-chain 2-hydroxyisobutyric acid (2-HIBA) has been associated with several metabolic diseases and lysine 2-hydroxyisobutyrylation seems to be a common eukaryotic as well as prokaryotic post-translational modification in proteins. In contrast, the underlying 2-HIBA metabolism has thus far only been detected in a few microorganisms, such as the betaproteobacterium Aquincola tertiaricarbonis L108 and the Bacillus group bacterium Kyrpidia tusciae DSM 2912. In these strains, 2-HIBA can be specifically activated to the corresponding CoA thioester by the 2-HIBA-CoA ligase (HCL) and is then isomerized to 3-hydroxybutyryl-CoA in a reversible and B 12-dependent mutase reaction. Here, we demonstrate that the actinobacterial strain Actinomycetospora chiangmaiensis DSM 45062 degrades 2-HIBA and also its precursor 2-methylpropane-1,2-diol via acetone and formic acid by employing a thiamine pyrophosphate-dependent lyase. The corresponding gene is located directly upstream of hcl, which has previously been found only in operonic association with the 2-hydroxyisobutyryl-CoA mutase genes in other bacteria. Heterologous expression of the lyase gene from DSM 45062 in E. coli established a 2-hydroxyisobutyryl-CoA lyase activity in the latter. In line with this, analysis of the DSM 45062 proteome reveals a strong induction of the lyase-HCL gene cluster on 2-HIBA. Acetone is likely degraded via hydroxylation to acetol catalyzed by a MimABCD-related binuclear iron monooxygenase and formic acid appears to be oxidized to CO 2 by selenium-dependent dehydrogenases. The presence of the lyase-HCL gene cluster in isoprene-degrading Rhodococcus strains and Pseudonocardia associated with tropical leafcutter ant species points to a role in degradation of biogenic short-chain ketones and highly branched organic compounds.

show abstract

Section: Discussionmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Organization Of the Lyase-hcl Gene Cluster In A Chiangmaienmentioning

confidence: 99%

See 2 more Smart Citations

Actinobacterial Degradation of 2-Hydroxyisobutyric Acid Proceeds via Acetone and Formyl-CoA by Employing a Thiamine-Dependent Lyase Reaction

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Kürzlich wurde von Chou et al in der Tat gezeigt, dass HACL die Acyloin-Kondensation zwischen Formyl-CoA und Aldehyd-Substraten katalysiert. [11] Diese Studie konzentrierte sich lediglich auf HACL und Formyl-CoA, während das Carboligationspotential von OXC mit Oxalyl-CoA als Donor unerforscht blieb.…”

unclassified

Oxalyl‐CoA Decarboxylase katalysiert die nukleophile ein‐Kohlenstoff‐Verlängerung von Aldehyden zu chiralen α‐Hydroxysäuren

2020

View full text Add to dashboard Cite

Die Synthese komplexer Moleküle aus erneuerbaren Kohlenstoffbausteinen ist ein zentrales Ziel einer nachhaltigen Wirtschaft. Hier untersuchten wir das biokatalytische Potential Thiamindiphosphat (ThDP)‐abhängiger Oxalyl‐CoA Decarboxylasen (OXC)/2‐Hydroxyacyl‐CoA Lyasen (HACL). Diese Enzyme katalysieren den Abbau von Acyl‐CoA‐Thioestern unter Bildung des C1‐Moleküls Formyl‐CoA. In der OXC/HACL Superfamilie kommen aber auch unspezifische Enzymvarianten vor, die C1‐Bausteine mit verschiedenen Aldehyden kondensieren und so 2‐Hydroxyacyl‐CoA‐Thioester bilden. Eine verbesserte OXC‐Variante mit zwei neu beschriebenen Enzymen – einer spezifischen Oxalyl‐CoA Synthetase und einer 2‐Hydroxyacyl‐CoA Thioesterase — wurde zu einer Kaskade kombiniert, die die Umsetzung von Oxalat und aromatischen Aldehyden zu (S)‐α‐Hydroxysäuren mit einem Enantiomerenüberschuss (ee) bis zu 99 % ermöglicht. Unsere Studie zeigt das Potential ThDP‐katalysierter C1‐Erweiterungsreaktionen in der nachhaltigen Synthese chiraler Bausteine.

show abstract

Oxalyl‐CoA Decarboxylase Enables Nucleophilic One‐Carbon Extension of Aldehydes to Chiral α‐Hydroxy Acids

2020

View full text Add to dashboard Cite

The synthesis of complex molecules from simple, renewable carbon units is the goal of a sustainable economy. Here we explored the biocatalytic potential of the thiamine‐diphosphate‐dependent (ThDP) oxalyl‐CoA decarboxylase (OXC)/2‐hydroxyacyl‐CoA lyase (HACL) superfamily that naturally catalyzes the shortening of acyl‐CoA thioester substrates through the release of the C1‐unit formyl‐CoA. We show that the OXC/HACL superfamily contains promiscuous members that can be reversed to perform nucleophilic C1‐extensions of various aldehydes to yield the corresponding 2‐hydroxyacyl‐CoA thioesters. We improved the catalytic properties of Methylorubrum extorquens OXC by rational enzyme engineering and combined it with two newly described enzymes—a specific oxalyl‐CoA synthetase and a 2‐hydroxyacyl‐CoA thioesterase. This enzymatic cascade enabled continuous conversion of oxalate and aromatic aldehydes into valuable (S)‐α‐hydroxy acids with enantiomeric excess up to 99 %.

show abstract

2-Hydroxyacyl-CoA lyase catalyzes acyloin condensation for one-carbon bioconversion

Cited by 65 publications

References 41 publications

Actinobacterial Degradation of 2-Hydroxyisobutyric Acid Proceeds via Acetone and Formyl-CoA by Employing a Thiamine-Dependent Lyase Reaction

Actinobacterial Degradation of 2-Hydroxyisobutyric Acid Proceeds via Acetone and Formyl-CoA by Employing a Thiamine-Dependent Lyase Reaction

Oxalyl‐CoA Decarboxylase katalysiert die nukleophile ein‐Kohlenstoff‐Verlängerung von Aldehyden zu chiralen α‐Hydroxysäuren

Oxalyl‐CoA Decarboxylase Enables Nucleophilic One‐Carbon Extension of Aldehydes to Chiral α‐Hydroxy Acids

Contact Info

Product

Resources

About