A synthetic CRISPR-Cas nuclease with expanded enzymatic activities DOI Open Access
Ylenia Jabalera, Igor Tascón, Sara Samperio

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Ноя. 12, 2023

Summary Clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonucleases have revolutionized biotechnology for their potential as programmable genome editors. Yet, most natural nucleases and variants limitations. Here, we report a fully synthetic CRISPR-associated (Cas) nuclease (α-synCas) designed by Ancestral Sequence Reconstruction (ASR) that displays set of robust distinct targeting properties, not found in any other known CRISPR-Cas Class 2 system. We show α-synCas is PAMless able to catalyse RNA-guided, specific cleavage dsDNA, ssDNA ssRNA. The enzyme also capable sequence-nonspecific degradation ssRNA following activation complementary targets. Furthermore, exhibits editing activity human cells bacteria. Cryo-electron microscopy structures ternary quaternary complexes provide framework understand the structural basis its expanded enzymatic activities. capability multimodal virtually nucleic acid sequence distinguishes promising new tool extend current CRISPR-based technologies.

Язык: Английский

A thiopyridine-bound mirror-image copper center in an artificial non-heme metalloenzyme DOI
Yoshitsugu Morita,

Hiroki Kubo,

Ryusei Matsumoto

и другие.

Journal of Inorganic Biochemistry, Год журнала: 2024, Номер 260, С. 112694 - 112694

Опубликована: Авг. 13, 2024

Язык: Английский

Процитировано

1
All-in-One CO2 Capture and Transformation: Lessons from Formylmethanofuran Dehydrogenases DOI
Olivier N. Lemaire, Tristan Wagner

Accounts of Chemical Research, Год журнала: 2024, Номер 57(24), С. 3512 - 3523

Опубликована: Ноя. 25, 2024

ConspectusCarbon-one-unit (C1) feedstocks are generally used in the chemical synthesis of organic molecules, such as solvents, drugs, polymers, and fuels. Contrary to dangerous polluting carbon monoxide mostly coming from fossil fuels, formate formamide attractive alternative for synthesis. As these currently mainly obtained oil industry, novel synthetic routes have been developed based on transformation greenhouse gas CO2. Such developments motivated by urgent need recycling, leading a sustainable future. The inert nature CO2 represents challenge chemists activate specifically convert molecule through an affordable efficient process. could be inspired biological activation, which highly specialized enzymes perform atmospheric fixation relatively abundant metal catalysts. In this Account, we describe discuss potential one most CO2-converting systems: formylmethanofuran dehydrogenase (abbreviated FMD).FMDs multienzymatic complexes found archaea that capture formyl group branched amine moiety methanofuran (MFR) cofactor. This overall reaction formyl−MFR production does not require ATP hydrolysis compared CO2-fixing microbes relying reductive Wood–Ljungdahl pathway, highlighting different operative mode saves cellular energy. FMD entry point hydrogenotrophic methanogenesis (H2 dependent or dependent) operates reverse other methanogenic pathways microbial metabolisms. Therefore, is key enzyme planetary cycle. After decades investigations, recent studies provided description structure, mechanism, electroreduction CO2, our laboratory has actively contributing.FMD "all-in-one" catalyzing redox-active coupled redox-neutral at two very cofactors where new C–H C–N bonds made. First, principle consisting exergonic reduction with endergonic condensation MFR resumed. Then, Account exposes molecular details active sites provides overview each catalytic mechanism. It also describes natural versatility electron-delivery modules fueling extends it possibilities using artificial systems electrodes.A perspective concludes how mechanistic applied produce CO2-based intermediates synthesize molecules. Indeed, its biochemical properties, opens opportunities generate molecules derivatives, all synthesizing compounds. Transferring knowledge acquired would provide coherent models can lead further development field biology bio-inspired chemistry large-scale conversion into building blocks

Язык: Английский

Процитировано

1
Designing Michaelases: exploration of novel protein scaffolds for iminium biocatalysis DOI Creative Commons
Alejandro Gran‐Scheuch, Stefanie Hanreich,

Iris Keizer

и другие.

Faraday Discussions, Год журнала: 2024, Номер 252, С. 279 - 294

Опубликована: Янв. 1, 2024

The study identified promising protein scaffolds for artificial enzyme development in iminium-ion catalysis, demonstrating activity and enantioselectivity abiological Michael addition reactions.

Язык: Английский

Процитировано

1
Excelzyme: A Swiss University-Industry Collaboration for Accelerated Biocatalyst Development DOI Creative Commons
Sumire Honda Malca, Peter Stockinger,

Nadine Duss

и другие.

CHIMIA International Journal for Chemistry, Год журнала: 2024, Номер 78(3), С. 108 - 117

Опубликована: Март 27, 2024

Excelzyme, an enzyme engineering platform located at the Zurich University of Applied Sciences, is dedicated to accelerating development tailored biocatalysts for large-scale industrial applications. Leveraging automation and advanced computational techniques, including machine learning, efficient can be generated in short timeframes. Toward this goal, Excelzyme systematically selects suitable protein scaffolds as foundation constructing complex libraries, thereby enhancing sequence structural biocatalyst diversity. Here, we describe applied workflows technologies well case study that exemplifies successful application workflow.

Язык: Английский

Процитировано

1
A resurrected ancestor of Cas12a expands target access and substrate recognition for nucleic acid editing and detection DOI
Ylenia Jabalera, Igor Tascón, Sara Samperio

и другие.

Nature Biotechnology, Год журнала: 2024, Номер unknown

Опубликована: Окт. 31, 2024

Язык: Английский

Процитировано

1
Artificial metalloenzymes enabled by combining proteins with hemin via protein refolding DOI
Jingping Ouyang, Zhenfang Zhang, René Hübner

и другие.

CHINESE JOURNAL OF CATALYSIS (CHINESE VERSION), Год журнала: 2024, Номер 67, С. 157 - 165

Опубликована: Дек. 1, 2024

Язык: Английский

Процитировано

1
Reactivity regulation for olefin metathesis-catalyzing ruthenium complexes with sulfur atoms at the terminal of 2-alkoxybenzylidene ligands DOI

Tsubasa Kinugawa,

Takashi Matsuo

Dalton Transactions, Год журнала: 2023, Номер 52(27), С. 9499 - 9508

Опубликована: Янв. 1, 2023

For regulating the olefin metathesis (OM) activity of Hoveyda-Grubbs second-generation complex (HG-II), structural modification benzylidene ligand is a useful strategy. This paper reports effect chalcogen atom placed at end group on catalytic properties HG-II derivatives, using complexes with thioether or ether component in (ortho-Me-E-(CH2)2O-styrene; E = S, O). Nuclear magnetic resonance and X-ray crystallographic analyses moiety (E S) proved (O,S)-bidentate trans-dichlorido coordination for complex. A stoichiometric exchange between produced corresponding an 86% yield, confirming higher stability than that HG-II. Despite bidentate chelation, exhibited OM activity, indicating exchangeability S-chelating olefinic substrate. The green solution color, characteristic was retained after S)-mediated reactions, high catalyst durability. Conversely, O) rapidly initiated reactions; however, it showed low In reactions conducted presence methanol, yields HG-II: S-coordination increased tolerance to methanol. coordinative (such as sulfur) terminal can precisely regulate reactivity derivatives.

Язык: Английский

Процитировано

3
Chemoenzymatic cascade reaction as a sustainable and scalable access to para-quinols DOI
Jan Samsonowicz‐Górski,

Anastasiia Hrunyk,

Anna Brodzka

и другие.

Green Chemistry, Год журнала: 2023, Номер 25(16), С. 6306 - 6314

Опубликована: Янв. 1, 2023

A novel protocol for the synthesis of para -quinols from inexpensive phenols based on a chemoenzymatic cascade reaction was developed.

Язык: Английский

Процитировано

2
A Review Towards Sustainable Analyte Detection: Biomimetic Inspiration in Biosensor Technology DOI
Pratistha Bhagat, Lata Sheo Bachan Upadhyay

Journal of Biotechnology, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 1, 2024

Язык: Английский

Процитировано

0
Using BpyAla to generate Copper Artificial Metalloenzymes: a catalytic and structural study DOI Creative Commons
Eva Klemencic, Richard C. Brewster, Hafız Muhammad Ali

и другие.

Опубликована: Окт. 30, 2023

Artificial metalloenzymes (ArMs) have emerged as a promising avenue in the field of biocatalysis, offering new reactivity. However, their design remains challenging due to limited understanding protein dynamics and how introduced cofactors alter scaffold structure. Here we present structures catalytic activity novel copper ArMs capable (R)- or (S)-stereoselective control, utilizing steroid carrier (SCP) scaffold. To incorporate 2,2’-Bipyridine (Bpy) into SCP, two distinct strategies were employed: either Bpy was an unnatural amino acid (2,2’-bipyridin-5-yl)alanine (BpyAla) using amber stop codon expression via bioconjugation bromomethyl-Bpy cysteine residues. The resulting proved be effective at catalysing enantioselective Friedel-Crafts reaction with SCP_Q111BpyAla achieving best selectivity enantioselectivity 72% ee (S). Interestingly, despite same scaffold, different attachment for residue (Q111) led switch enantiopreference ArM.

Язык: Английский

Процитировано

1