A5001133461- Microbial Metabolic Engineering and Bioproduction
- Protein purification and stability
- Biochemical Acid Research Studies
- Monoclonal and Polyclonal Antibodies Research
- Enzyme Catalysis and Immobilization
- Viral Infectious Diseases and Gene Expression in Insects
- Enzyme Structure and Function
- Protein Structure and Dynamics
- Glycosylation and Glycoproteins Research
- Advanced Thermodynamics and Statistical Mechanics
- Metabolism and Genetic Disorders
- Catalysis for Biomass Conversion
- Transgenic Plants and Applications
- Chemical Synthesis and Analysis
- Biofuel production and bioconversion
- RNA and protein synthesis mechanisms
- Various Chemistry Research Topics
- Peptidase Inhibition and Analysis
- Computational Drug Discovery Methods
- Innovative Microfluidic and Catalytic Techniques Innovation
- Biochemical and Molecular Research
- Signaling Pathways in Disease
- Alcoholism and Thiamine Deficiency
- Amino Acid Enzymes and Metabolism
- Biosimilars and Bioanalytical Methods
University College London
2016-2025
University College Lahore
2023
Biochemical Society
2003-2022
Engineering and Physical Sciences Research Council
2015-2022
Australian Centre for Robotic Vision
2022
Transnational Press London
2016
UCL Australia
2007-2011
University of Pennsylvania
2011
Institute of Structural and Molecular Biology
2009
Technical University of Denmark
2009
Abstract Flexible sites are potential targets for engineering the stability of enzymes. Nevertheless, success rate rigidifying flexible (RFS) strategy is still low due to a limited understanding how determine best mutation candidates. In this study, two parallel strategies were applied identify candidates within loops Escherichia coli transketolase (TK). The first was “back consensus mutations” approach, and second computational design based on ΔΔ G calculations in Rosetta. Forty-nine single...
Significance Incomplete understanding of the mechanisms epistasis between two or more substitutions in a protein can hamper protein-engineering strategies. With Escherichia coli transketolase as model, we explore epistatic interactions set stabilizing mutations from across different domains within structure. Surprisingly, not all pairwise effects distant surface and core regions were additive. Additionally, behaviors observed dependent on type stability measured. We found single that altered...
Significance Rigidifying flexible sites is a powerful method to improve enzyme stability. However, if the highly regions form active site, modifying them risks losing activity due activity–stability trade-off. We hypothesized here that outside site whose dynamics were correlated sites, would provide good targets for stabilizing mutations. To test this hypothesis, six variants constructed in 3M variant of Escherichia coli transketolase. The best had 10.8-fold improved half-life at 55 °C, and...
Despite recent advances in computational protein science, the dynamic behavior of proteins, which directly governs their biological activity, cannot be gleaned from sequence information alone. To overcome this challenge, we propose a framework that integrates peptide sequence, structure, and dynamics descriptors into machine learning algorithms to enhance predictive capabilities achieve improved prediction variant function. The resulting pipeline traditional structure with molecular...
Abstract Biocatalysis continues to emerge as a powerful technique for the efficient synthesis of optically pure pharmaceuticals that are difficult access via conventional chemistry. The power biocatalysis can be enhanced if two or more reactions achieved by single whole cell biocatalyst containing pathway designed de‐novo facilitate required synthetic sequence. enzymes transketolase (TK) and transaminase (TAm) respectively catalyze asymmetric carboncarbon bond formation amine group addition...
Abstract The engineering of enzymes with altered activity, specificity and stability, using directed evolution techniques that mimic on a laboratory timescale, is now well established. However, the general acceptance these methods as route to new biocatalysts for organic synthesis requires further improvement both ease-of-use also obtaining more significant changes in enzyme properties than currently possible. Recent advances library design, random mutagenesis, combined screening selection...
Chiral amino alcohols represent an important class of value-added biochemicals and pharmaceutical intermediates. Chemical routes to such compounds are generally step intensive, requiring environmentally unfriendly catalysts solvents. This work describes a multidisciplinary approach the rapid establishment biocatalytic chiral aminodiols taking original synthesis (2S,3S)-2-aminopentane-1,3-diol as specific example. An engineered variant Escherichi coli transketolase (D469T) was used for...
Transketolase mutants previously identified for use with the non-phosphorylated aldehyde propanal have been explored a series of linear and cyclic aliphatic aldehydes, excellent stereoselectivities observed.
Abstract Effective application of whole-cell devices in synthetic biology and biocatalysis will always require consideration the uptake molecules interest into cell. Here we demonstrate that AlkL protein from Pseudomonas putida GPo1 is an alkane import capable industrially relevant rates C 7 -C 16 n-alkanes. Without alkL expression, native E.coli n-alkane was rate-limiting step both bioconversion n-alkanes activation a biosensor by 10 11 alkanes. By coexpression as transporter plug-in,...
The analytical characterization of biopharmaceuticals is a fundamental step in the early stages development and prediction their behavior bioprocesses. Protein aggregation particular common issue as it affects all product development. In present work, we investigate stability kinetics A33Fab, therapeutically relevant humanized antibody fragment at wide range pH, ionic strength, temperature. We show that propensity A33Fab to aggregate under thermally accelerated conditions pH ionic-strength...
Over 8 million tonnes of sugar beet are grown annually in the UK. Sugar pulp (SBP) is main by-product processing which currently dried and sold as a low value animal feed. SBP rich source carbohydrates, mainly form cellulose pectin, including d-glucose (Glu), l-arabinose (Ara) d-galacturonic acid (GalAc). This work describes technical feasibility an integrated biorefinery concept for fractionation conversion these monosaccharides into value-added products. initially carried out by steam...
Neurofilament light chain (NfL) is an early nonspecific biomarker in neurodegenerative diseases and traumatic brain injury, indicating axonal damage. This work describes the detailed structural characterization of a selected primary calibrator with potential to be used future reference measurement procedure (RMP) development for accurate quantification NfL. As part described workflow, sequence, higher-order structure as well solvent accessibility, hydrogen-bonding profile were assessed under...
Beta-2-Glycoprotein I (β2GPI) is the main autoantigenic target of antiphospholipid syndrome (APS) with antibodies leading to clinical manifestations. There are two known structural isomers β2GPI, a J shape and circular shaped one. The transition between these structures incompletely understood, functional implications unknown. β2GPI substrate protease plasmin, which cleaves within fifth domain altered cellular binding. Very little currently regarding structure function this protein variant....