TY - JOUR
T1 - Enzyme-Loaded Nanoreactors Enable the Continuous Regeneration of Nicotinamide Adenine Dinucleotide in Artificial Metabolisms
AU - Jo, Seong-Min
AU - Wurm, Frederik R.
AU - Landfester, Katharina
N1 - Wiley deal
Funding Information:
This work is part of the MaxSynBio consortium, which was jointly funded by the Federal Ministry of Education and Research of Germany and the Max Planck Society. We thank Dr. Robert Graf (MPIP) for Si solid‐state NMR measurements. We thank Petra Räder (MPIP) for TGA measurements. We thank Dr. Young‐Su Jeong (Agency for Defense Development in South Korea) for helpful discussion about enzymatic reactions. Open access funding enabled and organized by Projekt DEAL. 29
Publisher Copyright:
© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH
PY - 2021/3/29
Y1 - 2021/3/29
N2 - Nicotinamide adenine dinucleotide (NAD) is an essential coenzyme for numerous biocatalytic pathways. While in nature, NAD+ is continuously regenerated from NADH by enzymes, all synthetic NAD+ regeneration strategies require a continuous supply of expensive reagents and generate byproducts, making these strategies unattractive. In contrast, we present an artificial enzyme combination that produces NAD+ from oxygen and water continuously; no additional organic substrates are required once a minimal amount pyruvate is supplied. Three enzymes, i.e., LDH, LOX, and CAT, are covalently encapsulated into a substrate-permeable silica nanoreactor by a mild fluoride-catalyzed sol–gel process. The enzymes retain their activity inside of the nanoreactors and are protected against proteolysis and heat. We successfully used NAD+ from the nanoreactors for the continuous production of NAD+ i) to sense glucose in artificial glucose metabolism, and ii) to reduce the non-oxygen binding methemoglobin to oxygen-binding hemoglobin. This latter conversion might be used for the treatment of Methemoglobinemia. We believe that this versatile tool will allow the design of artificial NAD+-dependent metabolisms or NAD+-mediated redox-reactions.
AB - Nicotinamide adenine dinucleotide (NAD) is an essential coenzyme for numerous biocatalytic pathways. While in nature, NAD+ is continuously regenerated from NADH by enzymes, all synthetic NAD+ regeneration strategies require a continuous supply of expensive reagents and generate byproducts, making these strategies unattractive. In contrast, we present an artificial enzyme combination that produces NAD+ from oxygen and water continuously; no additional organic substrates are required once a minimal amount pyruvate is supplied. Three enzymes, i.e., LDH, LOX, and CAT, are covalently encapsulated into a substrate-permeable silica nanoreactor by a mild fluoride-catalyzed sol–gel process. The enzymes retain their activity inside of the nanoreactors and are protected against proteolysis and heat. We successfully used NAD+ from the nanoreactors for the continuous production of NAD+ i) to sense glucose in artificial glucose metabolism, and ii) to reduce the non-oxygen binding methemoglobin to oxygen-binding hemoglobin. This latter conversion might be used for the treatment of Methemoglobinemia. We believe that this versatile tool will allow the design of artificial NAD+-dependent metabolisms or NAD+-mediated redox-reactions.
KW - UT-Hybrid-D
KW - encapsulation
KW - enzyme reactions
KW - nanoreactors
KW - nicotinamide adenine dinucleotide
KW - artificial metabolism
UR - http://www.scopus.com/inward/record.url?scp=85101652660&partnerID=8YFLogxK
U2 - 10.1002/anie.202012023
DO - 10.1002/anie.202012023
M3 - Article
SN - 1433-7851
VL - 60
SP - 7728
EP - 7734
JO - Angewandte Chemie (international edition)
JF - Angewandte Chemie (international edition)
IS - 14
ER -