Keeping the Distance: Activity Control in Solid-Supported Sucrose Phosphorylase by a Rigid α-Helical Linker of Tunable Spacer Length DOI Creative Commons
Chao Zhong, Anisha Vyas, Jakob Liu

et al.

ACS Catalysis, Journal Year: 2024, Volume and Issue: 14(22), P. 17090 - 17102

Published: Nov. 6, 2024

Enzyme immobilization into carrier materials has broad importance in biotechnology, yet understanding the catalysis of enzymes bound to solid surfaces remains challenging. Here, we explore surface effects on sucrose phosphorylase through a fusion protein approach. We immobilize enzyme via structurally rigid α-helical linker [EA3K]n tunable spacer length due variable number pentapeptide repeats used (n = 6, 14, 19). Molecular modeling and simulation approaches delineate conformational space sampled by each relative its His-tag cap for tethering. The population distribution conformers gets broader, with consequent shift enzyme-to-surface distance larger values (≤15 nm), as increases. Based temperature kinetic studies, obtain an energetic description enzyme-to-linker fusions solution Ni2+-chelate agarose. solid-supported involve distinct changes enthalpy–entropy partitioning within frame invariant Gibbs free energy activation (ΔG‡ ∼61 kJ/mol at 30 °C). entropic contribution (−TΔS‡) ΔG‡ increases length, from −16.4 linker-free +7.9 [EA3K]19 linked fusion. immobilized is indistinguishable catalytic properties solution, which behave identically regardless their linker. Enzymes positioned closer arguably experience higher degree molecular organization ("rigidification") that must relax additional uptake heat, compensated gain entropy. Increased thermostability these (up 2.8-fold) consistent proposed rigidification effect. Collectively, our study reveals parameters shows dependence surface-tethering fundamental insight here obtained, together successful extension principle different (nigerose phosphorylase), suggests linker-based control protein–surface can be engineering strategy optimize activity characteristics enzymes.

Language: Английский

Bottom‐Up Synthesized Glucan Materials: Opportunities from Applied Biocatalysis DOI Creative Commons
Chao Zhong, Bernd Nidetzky

Advanced Materials, Journal Year: 2024, Volume and Issue: 36(27)

Published: March 22, 2024

Linear d-glucans are natural polysaccharides of simple chemical structure. They comprised d-glucosyl units linked by a single type glycosidic bond. Noncovalent interactions within, and between, the d-glucan chains give rise to broad variety macromolecular nanostructures that can assemble into crystalline-organized materials tunable morphology. Structure design functionalization for diverse material applications largely relies on top-down processing derivatization naturally derived starting materials. The approach encounters critical limitations in efficiency, selectivity, flexibility. Bottom-up approaches synthesis offer different, often more precise, ways polymer structure control provide means functional diversification widely inaccessible routes polysaccharide processing. Here engineered enzymes (glycosyltransferases, glycoside hydrolases phosphorylases, glycosynthases) polymerization described use applied biocatalysis bottom-up assembly specific structures is shown. Advanced resulting polymeric products further shown their important role development sustainable bio-based circular economy discussed.

Language: Английский

Citations

4
Advances on hybrid modelling for bioprocesses engineering: insights into research trends and future directions from a bibliometric approach DOI Creative Commons

Juan Federico Herrera-Ruiz,

Javier Fontalvo, Oscar Andrés Prado-Rúbio

et al.

Results in Engineering, Journal Year: 2024, Volume and Issue: unknown, P. 103548 - 103548

Published: Nov. 1, 2024

Language: Английский

Citations

3
Pushing the boundaries of phosphorylase cascade reaction for cellobiose production I: Kinetic model development DOI Creative Commons

Alexander Sigg,

Mario Klimacek,

Bernd Nidetzky

et al.

Biotechnology and Bioengineering, Journal Year: 2023, Volume and Issue: 121(2), P. 580 - 592

Published: Nov. 20, 2023

Abstract One‐pot cascade reactions of coupled disaccharide phosphorylases enable an efficient transglycosylation via intermediary α‐ d ‐glucose 1‐phosphate (G1P). Such transformations have promising applications in the production carbohydrate commodities, including cellobiose for food and feed use. Several studies shown sucrose phosphorylase synthesis from sucrose, but boundaries on transformation efficiency that result kinetic thermodynamic characteristics individual enzyme are not known. Here, we assessed a step‐by‐step systematic fashion practical requirements model to describe at industrially relevant substrate concentrations up 600 mM glucose each. Mechanistic initial‐rate models two‐substrate (sucrose + phosphate → G1P fructose) (G1P phosphate) were needed additionally required expansion by terms inhibition, particular distinctive two‐site inhibition (from Cellulumonas uda ). Combined with mass action accounting approach equilibrium, gave excellent fit robust prediction full reaction time courses wide range activities as well concentrations, variable substoichiometric concentration phosphate. The thus provides essential engineering tool disentangle highly interrelated factors conversion reaction; it establishes necessary basis window operation calculations targeted optimizations toward different process tasks.

Language: Английский

Citations

6
Keeping the Distance: Activity Control in Solid-Supported Sucrose Phosphorylase by a Rigid α-Helical Linker of Tunable Spacer Length DOI Creative Commons
Chao Zhong, Anisha Vyas, Jakob Liu

et al.

ACS Catalysis, Journal Year: 2024, Volume and Issue: 14(22), P. 17090 - 17102

Published: Nov. 6, 2024

Enzyme immobilization into carrier materials has broad importance in biotechnology, yet understanding the catalysis of enzymes bound to solid surfaces remains challenging. Here, we explore surface effects on sucrose phosphorylase through a fusion protein approach. We immobilize enzyme via structurally rigid α-helical linker [EA3K]n tunable spacer length due variable number pentapeptide repeats used (n = 6, 14, 19). Molecular modeling and simulation approaches delineate conformational space sampled by each relative its His-tag cap for tethering. The population distribution conformers gets broader, with consequent shift enzyme-to-surface distance larger values (≤15 nm), as increases. Based temperature kinetic studies, obtain an energetic description enzyme-to-linker fusions solution Ni2+-chelate agarose. solid-supported involve distinct changes enthalpy–entropy partitioning within frame invariant Gibbs free energy activation (ΔG‡ ∼61 kJ/mol at 30 °C). entropic contribution (−TΔS‡) ΔG‡ increases length, from −16.4 linker-free +7.9 [EA3K]19 linked fusion. immobilized is indistinguishable catalytic properties solution, which behave identically regardless their linker. Enzymes positioned closer arguably experience higher degree molecular organization ("rigidification") that must relax additional uptake heat, compensated gain entropy. Increased thermostability these (up 2.8-fold) consistent proposed rigidification effect. Collectively, our study reveals parameters shows dependence surface-tethering fundamental insight here obtained, together successful extension principle different (nigerose phosphorylase), suggests linker-based control protein–surface can be engineering strategy optimize activity characteristics enzymes.

Language: Английский

Citations

1