scholarly journals A multi‐enzyme cascade for the biosynthesis of AICA ribonucleoside di‐ and triphosphate

ChemBioChem ◽  
2021 ◽  
Author(s):  
Lobna Eltoukhy ◽  
Christoph Loderer
Keyword(s):  
Enzyme Cascade

scholarly journals A Redox-Neutral, Two-Enzyme Cascade for the Production of Malate and Gluconate from Pyruvate and Glucose

2021 ◽  
Vol 11 (11) ◽  
pp. 4877
Author(s):  
Ravneet Mandair ◽  
Pinar Karagoz ◽  
Roslyn M. Bill
Keyword(s):  
Malate Dehydrogenase ◽  
Closed System ◽  
Glucose Dehydrogenase ◽  
Sulfolobus Solfataricus ◽  
Cofactor Regeneration ◽  
Triple Mutant ◽  
Thermococcus Kodakarensis ◽  
Platform Chemicals ◽  
Enzyme Cascade

A triple mutant of NADP(H)-dependent malate dehydrogenase from thermotolerant Thermococcus kodakarensis has an altered cofactor preference for NAD+, as well as improved malate production compared to wildtype malate dehydrogenase. By combining mutant malate dehydrogenase with glucose dehydrogenase from Sulfolobus solfataricus and NAD+/NADH in a closed reaction environment, gluconate and malate could be produced from pyruvate and glucose. After 3 h, the yield of malate was 15.96 mM. These data demonstrate the feasibility of a closed system capable of cofactor regeneration in the production of platform chemicals.

Efficient biotransformation and synergetic mechanism of dual-enzyme cascade reaction in nonreducing maltoheptaose synthesis

2021 ◽  
pp. 101066
Author(s):  
Luhua Zheng ◽  
Bo Jiang ◽  
Jingjing Chen ◽  
Tao Zhang ◽  
Xiaohong Gu ◽  
...  
Keyword(s):  
Cascade Reaction ◽  
Enzyme Cascade

scholarly journals A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Xu Tan ◽  
Sheng Zhang ◽  
Wei Song ◽  
Jia Liu ◽  
Cong Gao ◽  
...  
Keyword(s):  
Amino Acids ◽  
Specific Activity ◽  
Enantiomeric Excess ◽  
Efficient Production ◽  
Enzyme Cascade ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Rotation Strategy ◽  
Rate Limiting

AbstractIn this study, a four-enzyme cascade pathway was developed and reconstructed in vivo for the production of d-p-hydroxyphenylglycine (D-HPG), a valuable intermediate used to produce β-lactam antibiotics and in fine-chemical synthesis, from l-tyrosine. In this pathway, catalytic conversion of the intermediate 4-hydroxyphenylglyoxalate by meso-diaminopimelate dehydrogenase from Corynebacterium glutamicum (CgDAPDH) was identified as the rate-limiting step, followed by application of a mechanism-guided “conformation rotation” strategy to decrease the hydride-transfer distance d(C6HDAP−C4NNADP) and increase CgDAPDH activity. Introduction of the best variant generated by protein engineering (CgDAPDHBC621/D120S/W144S/I169P with 5.32 ± 0.85 U·mg−1 specific activity) into the designed pathway resulted in a D-HPG titer of 42.69 g/L from 50-g/L l-tyrosine in 24 h, with 92.5% conversion, 71.5% isolated yield, and > 99% enantiomeric excess in a 3-L fermenter. This four-enzyme cascade provides an efficient enzymatic approach for the industrial production of D-HPG from cheap amino acids.

Design of a Synthetic Enzyme Cascade for the in vitro Fixation of a C1 Carbon Source to a Functional C4 Sugar

2021 ◽  
Author(s):  
Samed Güner ◽  
Vanessa Wegat ◽  
André Pick ◽  
Volker Sieber
Keyword(s):  
Carbon Dioxide ◽  
Carbon Source ◽  
Greenhouse Gas ◽  
Value Added ◽  
Waste Stream ◽  
Enzyme Cascade ◽  
Sustainable Future ◽  
Value Added Products

Realizing a sustainable future requires intensifying the waste stream conversion, such as converting the greenhouse gas carbon dioxide into value-added products. In this paper, we focus on utilizing formaldehyde as...

scholarly journals Microfluidics Featuring Multilayered Hydrogel Assemblies Enable Real-Time NMR-Monitoring of Enzyme Cascade Reactions

2020 ◽  
Author(s):  
Nurdiana Nordin ◽  
lorenzo bordonali ◽  
Hossein Davoodi ◽  
Novindi Dwi Ratnawati ◽  
Gudrun Gygli ◽  
...  
Keyword(s):  
Real Time ◽  
Functional Performance ◽  
Cascade Reactions ◽  
Inhibitory Effect ◽  
Resonance Spectroscopy ◽  
Reaction Cell ◽  
Enzyme Cascade ◽  
Sensing Applications ◽  
Progress Curve ◽  
Michaelis Constants

Compartmentalized chemical reactions at the microscale are interesting from many perspectives including (multi)functional surfaces and biotechnology. Monitoring the molecular content as a measure of functional performance at these small scales is challenging. As a means to address this challenge, we leverage microtechnology and biocompatible materials to integrate a compact, reconfigurable reaction cell featuring electrochemical functionality with high-resolution nuclear magnetic resonance spectroscopy (NMR). We demonstrate the operation of this system by monitoring the activity of enzymes immobilized in chemically distinct layers within a multi-layered chitosan hydrogel assembly. As a benchmark, we observed the parallel activities of urease (Urs), catalase (Cat), and glucose oxidase (GOx) by recording NMR spectra to extract reagent and product concentrations in real-time. As a result, simultaneous monitoring of a cooperative enzymatic process (GOx + Cat) together with an independent process (Urs) is achieved. Using Michaelis-Menten progress curve analysis of the NMR data, kinetic data is extracted: in the case of GOx, the Michaelis constants (K<sub>M</sub>) are consistent with previous reports, while for Urs, deviations are observed, attributed to an inhibitory effect under our reaction conditions. The system therefore enables the construction of complex reaction cascades with spatial control, as would be interesting in, for example, metabolic engineering and multiplexed sensing applications.

scholarly journals Phase Separation Can Increase Enzyme Activity by Concentration and Molecular Organization

2020 ◽  
Author(s):  
William Peeples ◽  
Michael K. Rosen
Keyword(s):  
Phase Separation ◽  
Enzymatic Reaction ◽  
Reaction Rates ◽  
Quantitative Model ◽  
Mass Action ◽  
Molecular Organization ◽  
Scaffolding Proteins ◽  
Enzyme Cascade ◽  
Separation Threshold ◽  
Liquid Liquid Phase Separation

AbstractBiomolecular condensates concentrate macromolecules into discrete cellular foci without an encapsulating membrane. Condensates are often presumed to increase enzymatic reaction rates through increased concentrations of enzymes and substrates (mass action), although this idea has not been widely tested and other mechanisms of modulation are possible. Here we describe a synthetic system where the SUMOylation enzyme cascade is recruited into engineered condensates generated by liquid-liquid phase separation of multidomain scaffolding proteins. SUMOylation rates can be increased up to 36-fold in these droplets compared to the surrounding bulk, depending on substrate KM. This dependency produces substantial specificity among different substrates. Analyses of reactions above and below the phase separation threshold lead to a quantitative model in which reactions in condensates are accelerated by mass action and by changes in substrate KM, likely due to scaffold-induced molecular organization. Thus, condensates can modulate reaction rates both by concentrating molecules and by physically organizing them.

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