A bicyclic <i>S</i>-adenosylmethionine regeneration system applicable with different nucleosides or nucleotides as cofactor building blocks

Popadić, Désirée; Mhaindarkar, Dipali; Thai, Mike H N Dang; Hailes, Helen C.; Mordhorst, Silja; Andexer, Jennifer N.

doi:10.1039/d1cb00033k

Cited by 37 publications

(74 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In previous studies, polyphosphate kinases were widely used to make the conversions among AMP, ADP and ATP. For example, adenosine kinase (ADK), two family-2 polyphosphate kinases (PPK2-I and PPK2-II) are combined to make an ATP regeneration system which was used to synthesize SAM [ 31 , 32 ]. In another research, an AMP regeneration system was constructed by the adenosine kinase from Saccharomyces cerevisiae ( Sc ADK), a polyphosphate kinase from Acinetobacter johnsonii ( Aj PPK2), and a polyphosphate kinase from Sinorhizobium meliloti ( Sm PPK2).…”

Section: Discussionmentioning

confidence: 99%

Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins

Yang

et al. 2021

Biomolecules

View full text Add to dashboard Cite

Adenosine triphosphate (ATP) and S-adenosyl-L-methionine (SAM) are important intermediates that are widely present in living organisms. Large-scale preparation and application of ATP or SAM is limited by expensive raw materials. To lower the production costs for ATP/SAM, in this study we used strategies applying engineered multidomain scaffold proteins to synthesize ATP and SAM. An artificial scaffold protein containing CBM3 domain, IM proteins and CL-labeled proteins was assembled to form complex 1 for catalytic reactions to increase ATP production. The ATP synthesis system produced approximately 25 g/L of ATP with approximately 15 g/L of ADP and 5 g/L of AMP using 12.5 g/L of adenosine and 40 g/L of sodium hexametaphosphate reaction at 35 °C and a pH of 8.5 for 6 h. Based on the above ATP synthesis system, two CL-labeled methionine adenosyltransferases (CL9-MAT4 and CL9-MAT5) were applied to construct scaffold protein complex 2 to achieve SAM synthesis. Approximately 25 μg of MAT4 in a reaction system with 0.3 M MgCl2 catalyzed at 20 °C and a pH of 8 catalyzed 0.5 g/L of l-Met to produce approximately 0.9 g/L of SAM. Approximately 25 μg of MAT5 in a reaction system with 0.7 M MgCl2 catalyzed at 35 °C and a pH of 8 catalyzed 0.5 g/L of l-Met to produce approximately 1.2 g/L of SAM. Here, we showed that low-cost substrates can be efficiently converted into high-value additional ATP and SAM via multi-enzyme catalytic reactions by engineered multidomain scaffold proteins.

show abstract

Section: Discussionmentioning

confidence: 99%

Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins

Yang

et al. 2021

Biomolecules

View full text Add to dashboard Cite

show abstract

“…Overall, this results in a true SAM regeneration system (Figure 7A). After optimisation, this system achieves between 81 to 99 % conversion for the methylation of e. g. catechols and anthranilic acid with total turnover numbers (TTNs) of up to 200 [94,95] . In this cyclic regeneration system, homocysteine accumulates during the reactions.…”

Section: Sammentioning

confidence: 99%

Methyltransferases: Functions and Applications

et al. 2022

Self Cite

View full text Add to dashboard Cite

In this review the current state-of-the-art of S-adenosylmethionine (SAM)-dependent methyltransferases and SAM are evaluated. Their structural classification and diversity is introduced and key mechanistic aspects presented which are then detailed further. Then, catalytic SAM as a target for drugs, and approaches to utilise SAM as a cofactor in synthesis are introduced with different supply and regeneration approaches evaluated. The use of SAM analogues are also described. Finally O-, N-, C-and S-MTs, their synthetic applications and potential for compound diversification is given.

show abstract

“…Our data demonstrate that the proposed assay is a robust tool to measure MT activity with almost no limitation to the identity of the methylated substrate. In principle, the assay can be further expanded by the addition of an enzyme module that supplies SAM molecules to the transmethylation reaction starting from L‐methionine and ATP production [8c] …”

Section: Introductionmentioning

confidence: 99%

A High‐Throughput Continuous Spectroscopic Assay to Measure the Activity of Natural Product Methyltransferases

et al. 2022

View full text Add to dashboard Cite

Natural product methyltransferases (NPMTs) represent an emerging class of enzymes that can be of great use for the structural and functional diversification of bioactive compounds, such as the strategic modification of C-, N-, O-and Smoieties. To assess the activity and the substrate scope of the ever-expanding repertoire of NPMTs, a simple, fast, and robust assay is needed. Here, we report a continuous spectroscopic assay, in which S-adenosyl-L-methionine-dependent methylation is linked to NADH oxidation through the coupled activities of S-adenosyl-L-homocysteine (SAH) deaminase and glutamate dehydrogenase. The assay is highly suitable for a high-throughput evaluation of small molecule methylation and for determining the catalytic parameters of NPMTs under conditions that remove the potent inhibition by SAH. Through the modular design, the assay can be extended to match the needs of different aspects of methyltransferase cascade reactions and respective applications.

show abstract

A bicyclic S-adenosylmethionine regeneration system applicable with different nucleosides or nucleotides as cofactor building blocks

Abstract: The ubiquitous cofactor S-adenosylmethionine (SAM) is part of numerous biochemical reactions in metabolism, epigenetics, and cancer development. As methylation usually improves physiochemical properties of compounds relevant for pharmaceutical use, the...

Cited by 37 publications

References 68 publications

Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins

Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins

Methyltransferases: Functions and Applications

A High‐Throughput Continuous Spectroscopic Assay to Measure the Activity of Natural Product Methyltransferases

Contact Info

Product

Resources

About