Biosynthesis of macrocyclic peptides with C-terminal β-amino-α-keto acid groups by three different metalloenzymes DOI Creative Commons
Dinh Thanh Nguyen, Lingyang Zhu, Danielle L. Gray

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 30, 2023

Advances in genome sequencing and bioinformatics methods have identified a myriad of biosynthetic gene clusters (BGCs) encoding uncharacterized molecules. By mining genomes for BGCs containing prevalent peptide-binding domain used the biosynthesis ribosomally synthesized post-translationally modified peptides (RiPPs), we uncovered new class involving modifications installed by cytochrome P450, multi-nuclear iron-dependent non-heme oxidative enzyme (MNIO, formerly DUF692), cobalamin- radical

Language: Английский

Highly Stereoselective Biocatalytic One-Pot Synthesis of Chiral Saturated Oxygen Heterocycles by Integration of a Biosynthetic Heterocyclase into Multiple-Enzyme Cascades DOI Creative Commons

Theresa Roß-Taschner,

Sebastian Derra,

Jörg Stang

et al.

ACS Catalysis, Journal Year: 2024, Volume and Issue: 14(17), P. 13420 - 13428

Published: Aug. 23, 2024

The secondary metabolism is a rich source of enzymes with new synthetically attractive activities that have not yet been integrated into the toolbox biocatalysis. Chiral saturated oxygen heterocycles (CSOHs) are abundant structural elements natural products and other value-added compounds. We present biocatalytic method for synthesis CSOHs from readily accessible precursors combines an intramolecular oxa-Michael addition (IMOMA)-catalyzing cyclase (CYC) biosynthetic pathway alcohol dehydrogenases (ADHs) thioester-derivatizing enzymes. one-pot ADH–CYC reaction enables access to various tetrahydropyran (THP) tetrahydrofuran thioesters under control up four stereocenters. These convertible useful CSOH ketone, amide, aldehyde/alcohol, ester, carboxylic acid building blocks by chemical enzymatic means. extendibility more complex multienzyme cascades was demonstrated thioesterase reductase, allowing straightforward chemoenzymatic product (−)-civet, derivative, THP alcohol. integration IMOMA cyclases allows better exploitation high synthetic potential this group ring-forming expands repertoire pharmacologically relevant as highly selective versatile alternative. This approach will be adaptable wide range varying ADHs, cyclases, modifying

Language: Английский

Citations

1
Phosphorylation-condensation cascade for biocatalytic synthesis of C-nucleosides DOI Creative Commons

Andrej Ribar,

Martin Pfeiffer, Bernd Nidetzky

et al.

Chem Catalysis, Journal Year: 2024, Volume and Issue: unknown, P. 101127 - 101127

Published: Oct. 1, 2024

Language: Английский

Citations

1
Crystal structures and kinetic studies of a laboratory evolved aldehyde reductase explain the dramatic shift of its new substrate specificity DOI Creative Commons

Shruthi Sridhar,

Alberto Zavarise,

Tiila‐Riikka Kiema

et al.

IUCrJ, Journal Year: 2023, Volume and Issue: 10(4), P. 437 - 447

Published: May 25, 2023

The Fe 2+ -dependent E. coli enzyme FucO catalyzes the reversible interconversion of short-chain ( S )-lactaldehyde and )-1,2-propanediol, using NADH NAD + as cofactors, respectively. Laboratory-directed evolution experiments have been carried out previously phenylacetaldehyde substrate for screening catalytic activity with bulky substrates, which are very poorly reduced by wild-type FucO. These identified N151G/L259V double mutant (dubbed DA1472) most active variant this via a two-step evolutionary pathway, in each step consisted one point mutation. Here crystal structures DA1472 its parent D93 (L259V) reported, showing that these amino acid substitutions provide more space site, though they do not cause changes main-chain conformation. physiological series substituted derivatives were systematically quantified compared well corresponding point-mutation variants (N151G L259V). There is 9000-fold increase activity, when expressed k cat / K M values, substrate. structure complexed non-reactive analog (3,4-dimethoxyphenylacetamide) suggests mode binding group new combined structure–function studies therefore explain dramatic aldehyde substrates. comparisons also suggest why site replaced Zn able to support catalysis.

Language: Английский

Citations

2
Enzymes of the crotonase superfamily: Diverse assembly and diverse function DOI Creative Commons
Subhadra Dalwani, Rik K. Wierenga

Current Opinion in Structural Biology, Journal Year: 2023, Volume and Issue: 82, P. 102671 - 102671

Published: Aug. 4, 2023

The crotonase fold is generated by a framework of four repeats ββα-unit, extended two helical regions. active site superfamily (CS) enzymes located at the N-terminal end helix third repeat, typically being covered C-terminal helix. A major subset CS-enzymes catalyzes acyl-CoA-dependent reactions, allowing for diverse range acyl-tail modifications. Most these occur as trimers or hexamers (dimers trimers), but monomeric forms are also observed. common feature sites an oxyanion hole, formed peptide-NH hydrogen bond donors, which stabilises negatively charged thioester oxygen atom reaction intermediate. Structural properties and possible use biotechnological applications discussed.

Language: Английский

Citations

2
Biosynthesis of macrocyclic peptides with C-terminal β-amino-α-keto acid groups by three different metalloenzymes DOI Creative Commons
Dinh Thanh Nguyen, Lingyang Zhu, Danielle L. Gray

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 30, 2023

Advances in genome sequencing and bioinformatics methods have identified a myriad of biosynthetic gene clusters (BGCs) encoding uncharacterized molecules. By mining genomes for BGCs containing prevalent peptide-binding domain used the biosynthesis ribosomally synthesized post-translationally modified peptides (RiPPs), we uncovered new class involving modifications installed by cytochrome P450, multi-nuclear iron-dependent non-heme oxidative enzyme (MNIO, formerly DUF692), cobalamin- radical

Language: Английский

Citations

2