Altered Cofactor Preference of Thermostable StDAPDH by a Single Mutation at K159

International Journal of Molecular Sciences
Xiuzhen GaoMingfei Liu

Abstract

D-amino acid production from 2-keto acid by reductive amination is an attractive pathway because of its high yield and environmental safety. StDAPDH, a meso-diaminopimelate dehydrogenase (meso-DAPDH) from Symbiobacterium thermophilum, was the first meso-DAPDH to show amination of 2-keto acids. Furthermore, StDAPDH shows excellent thermostability compared to other meso-DAPDHs. However, the cofactor of StDAPDH is NADP(H), which is less common than NAD(H) in industrial applications. Therefore, cofactor engineering for StDAPDH is needed. In this study, the highly conserved cofactor binding sites around the adenosine moiety of NADPH were targeted to determine cofactor specificity. Lysine residues within a loop were found to be critical for the cofactor specificity of StDAPDH. Replacement of lysine with arginine resulted in the activity of pyruvic acid with NADH as the cofactor. The affinity of K159R to pyruvic acid was equal with NADH or NADPH as the cofactor, regardless of the mutation. Molecular dynamics simulations revealed that the large steric hindrance of arginine and the interaction of the salt bridge between NADH and arginine may have restricted the free movement of NADH, which prompted the formation of a stable active confo...Continue Reading

References

Sep 7, 1976·Biochemical and Biophysical Research Communications·H MisonoK Soda
Mar 1, 1987·Applied Biochemistry and Biotechnology·H K Chenault, G M Whitesides
Nov 1, 2003·Archives of Biochemistry and Biophysics·Hoon ChoHsin-Hsiung Tai
Aug 17, 2006·Journal of the American Chemical Society·Kavitha Vedha-PetersScott J Novick
Apr 27, 2007·Biotechnology Advances·Wenfang Liu, Ping Wang
Apr 22, 2010·Proteins·Kresten Lindorff-LarsenDavid E Shaw
Jan 21, 2015·Chemico-biological Interactions·Lilian González-SeguraRosario A Muñoz-Clares
Mar 12, 2015·Applied Microbiology and Biotechnology·Xiuzhen GaoHailiang Zhu
May 7, 2015·Acta Crystallographica. Section D, Biological Crystallography·Hironaga AkitaHaruhiko Sakuraba
Oct 30, 2015·Protein Engineering, Design & Selection : PEDS·J K B CahnF H Arnold
Sep 21, 2016·ACS Synthetic Biology·Jackson K B CahnFrances H Arnold
Aug 21, 2018·Frontiers in Microbiology·Hironaga AkitaToshihisa Ohshima
Feb 27, 2019·Journal of Chemical Information and Modeling·Xiuzhen GaoYuanda Song
Dec 14, 2019·Applied Microbiology and Biotechnology·Giorgia Letizia MarconeLoredano Pollegioni

❮ Previous
Next ❯

Methods Mentioned

BETA
deamination

Software Mentioned

PyMOL
GROMACS
GROMACS2018
genbox
Origin8

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.