Creative Enzymes is a leading professional in the field of enzyme ligand screening. We are specialized in rapid and accurate screening of substrates, inhibitors, and other ligands for specific enzymes, based on high-throughput experiments or computational simulation results. In other cases, we also provide screening of active enzymes against specific substrates or inhibitors, which would help to identify the functioning enzymes in a pathway or the biological target of a small molecule.
Generally, enzyme activity is measured under specific conditions, explicitly, at certain concentrations of the enzyme, substrate, cofactor, and in a fixed reaction environment. Activity screening, however, is often performed under constantly changing conditions, for that at least one component, either the substrate or the cofactor, never stays the same for the purpose of identifying the most active ones among a large pool of candidates. Therefore, proper design of the test and reasonable interpretation of the results is the key to effective screening tests. At Creative Enzymes, we have the knowledge and experience sourced from our top scientists, who support the screening test for a wide range of applications: identify the most active substrate, inhibitor screening for biotechnology, or even find the target enzyme of an inhibitor.
1. Substrate screening and identification:
Finding the most suitable substrate for an enzyme could be a tool for two aspects of one question: what reaction does an enzyme catalyze. The first fold of the question could be relatively easy to answer. We need to know the substrate, naturally occurred or synthesized in lab, that when mixed with the interested enzyme will show the maximal activity of this enzyme. The second fold of the question is, however, somewhat challenging to solve, but the answer will give more in-depth benefits—that is to discover the natural function of an enzyme by finding the molecules that work best with the enzyme. Although the purposes may be different, the technic to obtain the answers is the same. We rapidly measure the binding affinity and reaction rate of a large library of substrates. The most suitable substrate can then be identified as the ones that give the highest reposes.
In biomedical, chemical, and environmental applications, high-throughput approaches, such as microarray and automated testing, will reveal potential drug molecules, an efficient biosynthesis precursor, or the required building blocks for the target product. In more detailed biological research, scientists often need to understand the natural role of an enzyme. Although several databases can be used to align the protein or gene sequences to give a postulated function of a newly discovered enzyme, there is still a big gap in the numbers between experimentally identified enzymes and metabolites. It is supposed that many catalytic enzymes remain unknown in many biochemical pathways. Therefore, monitoring the reactions of ten thousands of substrate-enzyme pairs will give the key information of the natural function of the enzyme.
Creative Enzymes has established one of the largest substrate library, which does not just excel in the amount of the substrate, but also present a high variety of the substrate classes, including proteins, nucleotides, lipids, fatty acids, and other organic small molecules. We are able to give the identifications of the most likely substrate type, as well as comparison among similar substrates, such as phosphatase substrates, kinase substrates, and protease substrates. We will also choose the best test method for the specific enzyme that you may be interested in. Depending on the nature of the enzyme and the desired catalytic activity, the best method will give the most accurate results within a short timeframe and a reasonable cost. We have the capability of various test methods including radioautography, colorimetric, and flruorometric assays.
2. Inhibitor screening and design
Screening enzyme inhibitors is a necessary tool in life sciences and biotechnologies. The technic is especially helpful in drug discovery, but also used in more fundamental research and other industrial applications. The principle of inhibitor screening is similar to that of substrate screening, but may differ in certain ways. For some inhibitors, the process could be the same to substrate screening: a large number of inhibitors interact with the enzyme, and the magnitude of the binding affinity is indicated by the response of the reporting group, such as a fluorescent or radioactive group. For some competitive, non-competitive, and suicidal inhibitors, the screening could be even easier, using changes in the physical properties of the reaction interface such as refractive index or mass to rapidly detect the inhibition behavior. While in other cases, the assay could be much more complicated. The enzyme, substrate, and inhibitor need to be incubated in the same reaction mixture, and the inhibitory activity is measured by monitoring changes in the reaction rate. In some cases, the reaction is difficult to detect, an indirect method such as secondary reporter or a coupled enzyme assay has to be used.
Creative Enzymes has all the tools to implement any type of inhibitor screening. To ensure the most reliable results, we would like to understand your specific need, as detailed as possible. We will then choose the tests that best suit your request based on our expertise. In addition to the powerful screening platform, we also offer fast and economic in silico screening, if the biophysics of the enzyme is available. The virtual screening quickly looks through a large library of potential inhibitors and proposes the top candidates based on the desired inhibition. This is often used to narrow the searching range of the inhibitors, when a structural starting point is barely known to the researchers. We generally suggest a following experimental screening to confirm the result and to further focus the candidates on the top dozens of molecules.
3. Target enzyme identification
A different but similarly challenging task in biological research is to find the target enzyme of a known substrate or inhibitor. This is often seen in the search of the mode of action of a potential medicine, the effort to understand a mechanism of a disease, or the process to find the missing link in a metabolism pathway. In this case, the task can be considered as screening thousands of enzymes against a substrate or inhibitor, which could be as simple as a regular high-throughput screening or a short-term project that needs some investment and management. The main challenge of this type of screening is to keep all enzymes active at similar levels for an adequate period of time. In fact, almost every enzyme has its own optimal catalysis condition, and testing all of the candidate enzymes in one single condition is risky. Therefore, several conditions could be used for the same set of enzymes to cover the active conditions for all enzymes, or the candidate pool is sub-divide into smaller groups with similar properties before testing.
Creative Enzymes provides the highly reliable and customizable target identification tests by properly combining the skill of assay development and the power of advanced analytical instruments. We are happy to help you identify the target enzyme for an inhibitor, the natural biocatalyst of a certain reaction, or the most active enzyme that acts on a substrate. You will find the service of great value in pharmaceutical research and development, chemical processing, environmental chemistry, and other industrial applications.
4. Identification and screening of other enzyme ligands
Enzymes are sensitive biological macromolecules. Their catalytic activity could be greatly changed by small factors in the environment. Among all these factors, enzyme ligands are recognized, disproportionate to their significant levels of impact on enzyme activity. An enzymatic reaction could completely halt in the absence of a key cofactor, while the reaction rate could also be enhanced by hundreds of fold simply by adding a minimum amount of the right additive. These ligands or additives are so important to enzyme activities, yet may be difficult to study due to the vast range of possible chemical entities and interaction modes. For example, a nicotinamide adenine dinucleotide cofactor is used by the enzyme to simply transfer the electrons and protons, while a short peptide or metal ion could bind to the allosteric pocket to maintain the correct conformation for the enzyme to be active. Organic small molecules or electrolytes are sometimes necessary to the enzyme activity although not tightly bound to the enzyme. Other proteins or enzymes may also help to initiate and maintain the correct folding of an enzyme. The enzyme itself could be the trigger to activity at certain concentrations, for example, through auto-hydrolysis or oligomerization.
Creative Enzymes would like to help you test and determine the effect of various additives and possible ligands of an enzyme. We are capable of handling and properly testing all types of reaction additives or “helper molecules”. By using the proper additive, you could greatly improve the enzymatic reaction efficiency without having to perform expensive and time-consuming enzyme engineering and evolution. You may also consider testing the additives or ligands when troubleshooting an enzymatic reaction, which may not be working perfectly because of lacking the right additive. The nature of the service is highly technical, and Creative Enzymes promises to provide the most professional help in identification and screening of the right enzyme ligands.
Creative Enzymes enjoys the top reputation in screening tests of substrate, inhibitors, and enzymes. The test requires much knowledge and expertise in assay development and enzymology kinetics, which prevents many service providers from entering the field. Supported by outstanding enzymology researchers, Creative Enzymes is able to design and perform various screening tests for a variety of applications. Once your request is received, you can rely on Creative Enzymes to bring the package of solution and deliver the accurate results.
Welcome! For price inquiries, please feel free to contact us through the form below through the form on the left side. We will get back to you as soon as possible.