Enzymatic regeneration of ATP. I. Alternative routes

Adenosine-5'-triphosphate-dependent enzyme catalysed reactions are widespread in nature. Consequently, the enzymes involved have an intrinsic potential for use in syntheses of high value products. Although regeneration systems for ATP starting from adenosine-5'-diphosphate are available, certain limitations exist for both in vitro and in vivo applications requiring ATP regeneration from adenosine-5'-monophosphate, or adenosine. Following a short overview of the chemical and thermodynamic background, this Minireview focuses on emerging enzymes and methodologies for ATP regeneration. A large range of as yet unexploited reactions will be accessible with new, powerful, multistep ATP regeneration systems that use cheap phosphate donors and provide high longevity, compatibility, and robustness under process conditions. Their potential might go far beyond the direct use of ATP in enzymatic reactions; enzyme discovery, and engineering, as well as immobilisation strategies, will help to realise such systems.

show abstract

“…Of the many kinases described,48 only a few enzyme/phosphate donor combinations are commonly applied as regeneration systems (Table 2). 49…”

Section: Emerging Enzymes and Phosphate Donors Used For In Vitro Amentioning

confidence: 99%

Emerging Enzymes for ATP Regeneration in Biocatalytic Processes

2015

View full text Add to dashboard Cite

show abstract

“…Each of the assays was reproducible to within less than &2%. Additional details about experimental techniques are available (Langer, 1974 ).…”

Section: Analysesmentioning

confidence: 99%

Enzymatic regeneration of ATP: II. Equilibrium studies with acetate kinase and adenylate kinase

et al. 1977

Self Cite

View full text Add to dashboard Cite

Equilibria of reactions catalyzed by acetate kinase and adenylate kinase were studied experimentally and theoretically. Nucleotide conversions in excess of 90% were obtained with reactants (acetyl phosphate and one nucleotide) in stoichiometric proportion for ATP regeneration from either ADP via acetate kinase or AMP via the coupled enzyme system. Observed equilibrium constants, measured as a function of [Mg*+] at pH 7.4, ranged from about 1 to 9 for the reaction catalyzed by adenylate kinase and from about 50 to 400 for the reaction catalyzed by acetate kinase. These results agreed well with prediction of a theoretical model for the multiple equilibria between all species present in solution. Theoretical calculations showed that magnesium ion complexes and totally dissociated anions are the predominant species in solution at pH 7 and [Mg2+] F 10-3 M. SCOPEProspects for cell free enzyme cataIyzed syntheses on an industrial scale depend upon development of a practical process for regenerating adenosine 5'-triphosphate (ATP) from adenosine 5'-diphosphate (ADP) or adenosine 5'-monophosphate (AMP). A promising enzymatic route involves two reactions catalyzed by phosphotransferasesAcetate where acetyl phosphate (AcP) is the ultimate phosphate donor, and acetate (Ac) is the by-product. Both enzymes require a divalent cation (preferably Mg2+) for activity. Reaction (2) suffices for regeneration from ADP; both reactions are required for regeneration from AMP.In this paper we report equilibrium studies of reactions (1) and (2). Assays were developed to measure the concentration of each individual reactant and product in the presence of all other components and both enzymes. Observed equilibrium constants were measured for each Correspondence conrerning this paper should be addressed to C . K. experiments were conducted were selected to fall within the range where the enzymes retained substantial catalytic activity. The equilibrium composition of nucleotides was measured for ATP regeneration from AMP or ADP at pH 7.4 with the total concentrations of adenosine and magnesium held constant while acetyl phosphate concentration was varied.Under typical reaction conditions, each component may exist in a variety of forms, including totally dissociated anions, partially protonated species, divalent metal ion (Mg2+) complexes, and, under certain conditions, monovalent metal ion complexes. Reactions (1) and (2) therefore constitute a complicated network of reactions among a large variety of species, none of which is individually accessible to direct concentration meawrement.A theoretical model is presented for the multide equilibria between individual species; it permits calculation of the equilibrium comnosition given an arbitrary feed to a reactor. In developing the model it was necessary to determine which species should be incorporated 3nd to select from the literature a consistent set of stability and ionization constants which gave good agreement between prediction and data.AlChE Journal (Vol. 23, No. 1) January, 1977Page 1 C...

show abstract

“…Nature's way to enable phosphate ester formation makes use of ATP‐dependent kinases, that are rather substrate‐specific enzymes. Suitable cofactor recycling is necessary to reduce cost (Faber, ; Langer et al, ; Pollak et al, ; Shih and Whitesides, ). Despite the modest efficiency of ATP‐recycling, several preparative‐scale processes have been developed using hexokinase and glycerol kinase, although only sugar‐like molecules and derivatives of glycerol were well accepted (Chenault et al, ; Crans and Whitesides, ,; Drueckhammer and Wong, ).…”

Section: Introductionmentioning

confidence: 99%

Investigation of acid phosphatase variants for the synthesis of phosphate monoesters

Tasnádi

Zechner

Hall

et al. 2017

Biotech & Bioengineering

View full text Add to dashboard Cite

The major drawback of using phosphatases for transphosphorylation reactions lies in product depletion caused by the natural hydrolytic activity of the enzymes. Variants of PhoC-Mm from Morganella morganii and NSAP-Eb from Escherichia blattae were studied for their ability to maintain a high product level in the transphosphorylation of various primary alcohols. A single amino acid exchange delivered phosphatase variant PhoC-Mm G92D, which was able to catalyze the phosphorylation of primary alcohols without any major hydrolysis of the formed phosphate esters. The mutation mostly improved the affinity of the enzyme for alcohols, while rate constants of transphosphorylation and hydrolysis were decreased, overall resulting in a superior catalytic efficiency in transphosphorylation compared to hydrolysis. The presence of residual substrate alcohol at a given concentration was crucial to suppress phosphate ester hydrolysis. The present work extends the synthetic applicability of phosphatase variants beyond the previously reported nucleosides and allows preparative-scale production of various primary phosphate esters (yields up to 42%) with high enzyme productivity (TONs up to ∼66,000). Biotechnol. Bioeng. 2017;114: 2187-2195. © 2017 Wiley Periodicals, Inc.

show abstract

Enzymatic regeneration of ATP. I. Alternative routes

Cited by 65 publications

References 52 publications

Emerging Enzymes for ATP Regeneration in Biocatalytic Processes

Emerging Enzymes for ATP Regeneration in Biocatalytic Processes

Enzymatic regeneration of ATP: II. Equilibrium studies with acetate kinase and adenylate kinase

Investigation of acid phosphatase variants for the synthesis of phosphate monoesters

Contact Info

Product

Resources

About