Second generation engineering of transketolase for polar aromatic aldehyde substrates

Enzyme and Microbial Technology
Panwajee PayongsriPaul A Dalby

Abstract

Transketolase has significant industrial potential for the asymmetric synthesis of carboncarbon bonds with new chiral centres. Variants evolved on propanal were found previously with nascent activity on polar aromatic aldehydes 3-formylbenzoic acid (3-FBA), 4-formylbenzoic acid (4-FBA), and 3-hydroxybenzaldehyde (3-HBA), suggesting a potential novel route to analogues of chloramphenicol. Here we evolved improved transketolase activities towards aromatic aldehydes, by saturation mutagenesis of two active-site residues (R358 and S385), predicted to interact with the aromatic substituents. S385 variants selectively controlled the aromatic substrate preference, with up to 13-fold enhanced activities, and KM values comparable to those of natural substrates with wild-type transketolase. S385E even completely removed the substrate inhibition for 3-FBA, observed in all previous variants. The mechanisms of catalytic improvement were both mutation type and substrate dependent. S385E improved 3-FBA activity via kcat, but reduced 4-FBA activity via KM. Conversely, S385Y/T improved 3-FBA activity via KM and 4-FBA activity via kcat. This suggested that both substrate proximity and active-site orientation are very sensitive to mutation. Compa...Continue Reading

References

Jul 10, 1998·Biochimica Et Biophysica Acta·U Schörken, G A Sprenger
Jul 17, 1998·Current Opinion in Chemical Biology·W D Fessner
Jan 30, 1999·The International Journal of Biochemistry & Cell Biology·G SchenkP F Nixon
Feb 19, 2002·Current Opinion in Biotechnology·W D Fessner, V Helaine
Dec 10, 2003·Current Opinion in Biotechnology·Michael Breuer, Bernhard Hauer
Nov 1, 1995·Acta Crystallographica. Section D, Biological Crystallography·J LittlechildH Watson
Apr 15, 2006·Applied Microbiology and Biotechnology·Anne K Samland, Georg A Sprenger
Sep 11, 2007·Journal of Biotechnology·Edward G HibbertPaul A Dalby
Mar 18, 2008·Journal of Biotechnology·Edward G HibbertPaul A Dalby
Sep 10, 2008·The Journal of Organic Chemistry·Dieter Enders, Arun A Narine
Jun 9, 2009·Protein Engineering, Design & Selection : PEDS·Janahan ParamesvaranPaul A Dalby
Nov 17, 2009·Accounts of Chemical Research·Pilar HoyosPablo Domínguez de María
Mar 1, 2011·Current Opinion in Biotechnology·Verena ReschWolfgang Kroutil
Mar 23, 2011·Chemical Reviews·Margarita BrovettoGustavo A Seoane
Dec 14, 2011·Journal of Biotechnology·John StraffordPaul A Dalby
Sep 25, 2012·Chembiochem : a European Journal of Chemical Biology·Dong YiWolf-Dieter Fessner
Oct 20, 2012·Organic & Biomolecular Chemistry·Panwajee PayongsriPaul A Dalby

❮ Previous
Next ❯

Citations

Oct 22, 2016·Scientific Reports·Pierre E AffaticatiPaul A Dalby
Feb 2, 2017·Scientific Reports·Haoran YuPaul A Dalby
Dec 12, 2018·Proceedings of the National Academy of Sciences of the United States of America·Haoran Yu, Paul A Dalby

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.