Many successful stories in enzyme engineering are based on the creation of randomized diversity in large mutant libraries, containing millions to billions of enzyme variants. Methods that enabled their evaluation with high throughput are dominated by spectroscopic techniques due to their high speed and sensitivity. A large proportion of studies relies on fluorogenic substrates that mimic the chemical properties of the target or coupled enzymatic assays with an optical read-out that assesses the desired catalytic efficiency indirectly. The most reliable hits, however, are achieved by screening for conversions of the starting material to the desired product. For this purpose, functional group assays offer a general approach to achieve a fast, optical read-out. They use the chemoselectivity, differences in electronic and steric properties of various functional groups, to reduce the number of false-positive results and the analytical noise stemming from enzymatic background activities. This review summarizes the developments and use of functional group probes for chemoselective derivatizations, with a clear focus on screening for enzymatic activity in protein engineering.
The application of flour is determined by the composition of its starch and storage proteins. Previously isolated diploid wheat is known to be amylose-free and possesses the same amylopectin structure as the wild-type. To reveal its characteristics, starch, protein, lipid, fiber, gluten, and allergen contents and rheological properties were analyzed and compared to its parental wild-type diploid wheat and commercially available hexaploid wheats.
D-Allulose 3-epimerase catalyzes C-3 epimerization between D-fructose and D-allulose was found in strain M30. The enzyme gene was cloned, and its recombinant enzyme and the mutant variants were expressed in Using the information of the sequence and model structure, we succeed in the improvement of melting temperature for the enzyme without significant loss of the enzyme activity by protein engineering method. The melting temperatures were increased by 2.
Department of Cardiology, College of Medicine, Southwest Jiaotong University, Chengdu Cardiovascular Disease Research Institute, The Third People's Hospital of Chengdu, Chengdu, Sichuan, People's Republic of China.
Background: Increased levels of remnant cholesterol (RC) and inflammation are linked to higher risks of atherosclerotic cardiovascular disease. Whether a combination of C-reactive protein (CRP) and RC improves the predictive ability for evaluating the severity of coronary artery lesions remains unknown.
Methods: A total of 1675 patients with coronary artery disease were stratified according to the Synergy Between Percutaneous Coronary Intervention (SYNTAX) score (SYNTAX score ≤22 versus SYNTAX score >22).
Mussels and tubeworms have evolved similar adhesive systems to cope with the hydrodynamics of intertidal environments. Both secrete adhesive proteins rich in DOPA, a post-translationally modified amino acid playing essential roles in their permanent adhesion. DOPA is produced by the hydroxylation of tyrosine residues by tyrosinase enzymes, which can also oxidize it further into dopaquinone.
Polyethylene Terephthalate (PET) is a petroleum-based plastic polymer that, by design, can last decades, if not hundreds of years, when released into the environment through plastic waste leakage. In the pursuit of sustainable solutions to plastic waste recycling and repurposing, the enzymatic depolymerization of PET has emerged as a promising green alternative. However, the metabolic potential of the resulting PET breakdown molecules, such as the two-carbon (C2) molecule ethylene glycol (EG), remains largely untapped.
