Application of Transaminases

Höhne, Matthias; Bornscheuer, Uwe T.

doi:10.1002/9783527639861.ch19

Cited by 23 publications

(12 citation statements)

References 100 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 2 − 7 Consequently, they are also called amine transaminases in the literature. 3 , 4 , 8 , 9 ωTAs have received much attention in the last decade as they can be used as biocatalysts for the production of chiral amines. 10 − 18 One example of such a biocatalytically useful ωTA is the ( S )-selective ωTA (( S )-ωTA) from Chromobacterium violaceum (Cv-ωTA).…”

Section: Introductionmentioning

confidence: 99%

Quantum Chemical Study of Dual-Substrate Recognition in ω-Transaminase

2017

View full text Add to dashboard Cite

ω-Transaminases are attractive biocatalysts for the production of chiral amines. These enzymes usually have a broad substrate range. Their substrates include hydrophobic amines as well as amino acids, a feature referred to as dual-substrate recognition. In the present study, the reaction mechanism for the half-transamination of l-alanine to pyruvate in (S)-selective Chromobacterium violaceum ω-transaminase is investigated using density functional theory calculations. The role of a flexible arginine residue, Arg416, in the dual-substrate recognition is investigated by employing two active-site models, one including this residue and one lacking it. The results of this study are compared to those of the mechanism of the conversion of (S)-1-phenylethylamine to acetophenone. The calculations suggest that the deaminations of amino acids and hydrophobic amines follow essentially the same mechanism, but the energetics of the reactions differ significantly. It is shown that the amine is kinetically favored in the half-transamination of l-alanine/pyruvate, whereas the ketone is kinetically favored in the half-transamination of (S)-1-phenylethylamine/acetophenone. The calculations further support the proposal that the arginine residue facilitates the dual-substrate recognition by functioning as an arginine switch, where the side chain is positioned inside or outside of the active site depending on the substrate. Arg416 participates in the binding of l-alanine by forming a salt bridge to the carboxylate moiety, whereas the conversion of (S)-1-phenylethylamine is feasible in the absence of Arg416, which here represents the case in which the side chain of Arg416 is positioned outside of the active site.

show abstract

Section: Introductionmentioning

confidence: 99%

Quantum Chemical Study of Dual-Substrate Recognition in ω-Transaminase

2017

View full text Add to dashboard Cite

show abstract

“…Depending upon their substrate scope, transaminases can be divided into α‐, ω‐ and amine transaminases . α‐Transaminases convert substrates with the amino group in an alpha position to the carboxylate.…”

Section: Introductionmentioning

confidence: 99%

Structural and biochemical characterization of the dual substrate recognition of the (R)‐selective amine transaminase from Aspergillus fumigatus

et al. 2014

Self Cite

View full text Add to dashboard Cite

Chiral amines are important precursors for the pharmaceutical and finechemical industries. Because of this, the demand for enantiopure amines is currently increasing. Amine transaminases can produce a large spectrum of chiral amines in the (R)-or (S)-configuration, depending on their substrate scope and stereo-preference, by converting a prochiral ketone into the chiral amine while using alanine as the amine donor producing pyruvate as an a-keto acid product. In order to guide the protein engineering of transaminases to improve substrate specificity and enantioselectivity, we carried out a crystal structure analysis at 1.6A resolution of the (R)-amine transaminase from Aspergillus fumigatus with the bound inhibitor gabaculine. This revealed that Arg126 has an important role in the dual substrate recognition of this enzyme because mutating this residue to alanine reduced substantially the ability of the enzyme to use pyruvate as an amino acceptor. DatabaseCoordinates and structure factors have been deposited with the Protein Data Bank under accession code 4UUG.

show abstract

“…The ability to install amine groups, often in an enantioselective manner, is highly attractive, and when used to synthesize expensive products using relatively cheap amine donors such as isopropylamine, these methods can also become more economical. Although ω-transaminases are reported to have some limitations, such as low thermotolerance, substrate and product inhibition, and equilibrium-controlled yields, complementary methods have been developed to address these issues ( 7 , 10 – 12 ). In addition, there are a number of X-ray structures available for ω-TAs, and despite having highly conserved tertiary and quaternary structures, the enzymes exhibit conformational changes upon binding, and this conformational heterogeneity has been reported to make significant contributions to substrate specificity ( 13 ).…”

Section: Introductionmentioning

confidence: 99%

A β-Alanine Catabolism Pathway Containing a Highly Promiscuous ω-Transaminase in the 12-Aminododecanate-Degrading Pseudomonas sp. Strain AAC

Wilding

Peat

Newman

et al. 2016

Appl Environ Microbiol

View full text Add to dashboard Cite

We previously isolated the transaminase KES23458 from Pseudomonas sp. strain AAC as a promising biocatalyst for the production of 12-aminododecanoic acid, a constituent building block of nylon-12. Here, we report the subsequent characterization of this transaminase. It exhibits activity with a broad substrate range which includes α-, β-, and ω-amino acids, as well as α,ω-diamines and a number of other industrially relevant compounds. It is therefore a prospective candidate for the biosynthesis of a range of polyamide monomers. The crystal structure of KES23458 revealed that the protein forms a dimer containing a large active site pocket and unusual phosphorylated histidine residues. To infer the physiological role of the transaminase, we expressed, purified, and characterized a dehydrogenase from the same operon, KES23460. Unlike the transaminase, the dehydrogenase was shown to be quite selective, catalyzing the oxidation of malonic acid semialdehyde, formed from β-alanine transamination via KES23458. In keeping with previous reports, the dehydrogenase was shown to catalyze both a coenzyme A (CoA)-dependent reaction to form acetyl-CoA and a significantly slower CoA-independent reaction to form acetate. These findings support the original functional assignment of KES23458 as a β-alanine transaminase. However, a seemingly well-adapted active site and promiscuity toward unnatural compounds, such as 12-aminododecanoic acid, suggest that this enzyme could perform multiple functions for Pseudomonas sp. strain AAC.IMPORTANCE We describe the characterization of an industrially relevant transaminase able to metabolize 12-aminododecanoic acid, a constituent building block of the widely used polymer nylon-12, and we report the biochemical and structural characterization of the transaminase protein. A physiological role for this highly promiscuous enzyme is proposed based on the characterization of a related gene from the host organism. Molecular dynamics simulations were carried out to compare the conformational changes in the transaminase protein to better understand the determinants of specificity in the protein. This study makes a substantial contribution that is of interest to the broad biotechnology and enzymology communities, providing insights into the catalytic activity of an industrially relevant biocatalyst as well as the biological function of this operon.

show abstract

Application of Transaminases

Cited by 23 publications

References 100 publications

Quantum Chemical Study of Dual-Substrate Recognition in ω-Transaminase

Quantum Chemical Study of Dual-Substrate Recognition in ω-Transaminase

Structural and biochemical characterization of the dual substrate recognition of the (R)‐selective amine transaminase from Aspergillus fumigatus

A β-Alanine Catabolism Pathway Containing a Highly Promiscuous ω-Transaminase in the 12-Aminododecanate-Degrading Pseudomonas sp. Strain AAC

Contact Info

Product

Resources

About