Case Study: Improving Biofuels and Renewable Chemicals Production Through AI Driven Enzyme Engineering
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CD Biosynsis is a company specialised in enzyme-directed evolution services that provide customers with rapid and systematic enzyme-directed evolution services with the help of a platform using high-throughput screening technology as well as directed evolution methods. The technology we used for enzyme evolution based on genome engineering. By carrying out high-throughput random mutagenesis and high-throughput screening of a large number of enzyme molecules that have been overwhelmed to evolve the enhancement of enzyme catalytic function, the enzyme target substrate specificity, and the stability and tolerance of enzyme activity to organic solvents and other harsh environments can be achieved. While employing the enzyme function genetically encoded biosensors (GEBs) technologies, rapid identification of enzyme variants with new substrate scope or new reactivity can efficiently be made.
Directed evolution for enzyme development in biocatalysis (Serena G. and Patrice S., 2021)
Enzyme screening and directed evolution services, besides enzyme modification, enzyme immobilisation and process development and scale-up production of chemical-enzyme reactions will further streamline the research and development cycle for enterprises, meanwhile effectively reducing the failure rate of development, and help customers to break through industrial application and biopharmaceutical bottleneck.
Moreover, we also apply artificial intelligence technologies, such as machine learning to optimise directed evolution to enhance its efficiency, fidelity and performance. We are using the best of modern science to provide customers with quality and tailor-made solutions through impeccable enzyme engineering strategies to aid in the development of biomedicine and industrial applications.
Enzyme Directed Evolution Service is a technology that improves enzyme performance by simulating the natural evolution process. The methods we can provide are as follows:
Random mutation
This is one of the most common techniques in directed evolution. By randomly introducing genetic mutations to generate a large library of mutant enzymes, enzymes with desired characteristics are screened out.
Site-directed saturation mutation
This method is to make all possible individual amino acid substitutions at specific amino acid positions to systematically study the impact of each amino acid on enzyme activity.
Combinatorial library construction
Multiple mutation combinations are generated through different mutation strategies to form a library to find the best enzyme variant in subsequent screening
Truncated library
Study the impact of these regions on enzyme function by deleting part of the enzyme sequence.
Promoter evolution and screening
Evolving promoter sequences to increase gene expression levels and thereby improve enzyme production efficiency.
Random Insertion/Deletion Mutation
6.This method can produce mutant enzyme sets that contain unnatural amino acids, thereby increasing the functional diversity of the enzyme.
BAC/YAC modification
Genetic engineering done through bacterial artificial chromosomes (BAC) or yeast artificial chromosomes (YAC) for gene expression and stability.
These separated services are the foundational data of the Enzyme Directed Evolution Service, which aims to significantly improve and optimize enzyme performance through a variety of mutations and screens.
Enzyme Directed Evolution Service is a method of enhancing the performance of enzymes that mainly concentrates on the establishment of enzyme mutation Library construction and high-throughput screening. The process of this service is introduced below:
Construction of mutation library
High-throughput screening
Optimization and verification
Offer custom tailored services
Via the above process, we will offer customers faster and more customised enzyme-directed evolution services facilitating new drug-research and -development, as well as biotechnological applications.
Case 1: Improving enzyme tolerance in organic solvents
Project background:
A pharma company is developing a method to synthetise a drug by using an enzyme that is stable in the presence of high concentrations of organic solvents. In this way, they can ensure a more stable and efficient process, with improved safety in terms of handling and waste disposal. However, current enzymes performing this type of process function only at low concentrations of organic solvents so far, which entails higher production costs and instability.
Solution:
By directed evolution technology (a strategy of random mutation and selection), we constructed a mutant library and found the best one through high-throughput screening. This enzyme could work stably in high concentrations of organic solvents.
Result:
Following three rounds of directed evolution the activity of the gained enzyme was 6 times higher than in the wild type and its stability in organic solvents was drastically increased. This helps the company use cheaper organic solvents for drug synthesis while retaining the quality of the product.
Case 2: Enhance the catalytic ability of natural enzymes on non-natural substrates
Project background:
At a biotechnology company, they are developing a new enzyme that would be effective in catalysing the transformation of unnatural substrates. However, due to the low catalytic efficiency of existing natural enzymes on these substrates, they would not have such an application.
Solution:
To achieve this goal, we employed directed evolution technology by iteratively mutating a natural enzyme and screening for more efficient enzymes that catalyse non-natural substrates. We created several mutation libraries, and we screened them using our unique high-throughput screening technology.
Result:
By modifying the amino acids in its active centre, Delaney increased the enzyme's activity by 80 per cent, relative to the natural enzyme. On the one hand, the improved enzyme has been able to catalyse substrates that are outside of its natural range. On the other hand, it has allowed the company to open up new market opportunities.
Case 3: Improving the thermal stability of the enzyme
Project background:
Overall, a food processing company will develop an improved food processing procedure and product quality by using an enzyme that is more stable at high temperature. Specifically, it is currently almost impossible to efficiently improve food processing procedure and product quality because existing favourozyme excreted by fusarium is unstable at high temperature, which leads to a production interruption and product quality reduction during the food processing.
Solution:
By using the technology of directed evolution, we made alterations to the enzyme by using randomised mutation and screening, and constructed the mutant library. Through the screening of several rounds, the final enzyme remained active under high temperatures.
Result:
By performing several cycles of directed evolution, the research team increased the enzyme's thermal stability and activity five-fold, giving the company the leeway to use higher temperatures during food processing, which improves production efficiency and product quality.
These examples illustrate this strong ability of directed evolution technology to boost enzyme performance and help clients to solve affairs and achieve their needs.
The application areas and uses of Enzyme Directed Evolution service can be summarised as follows:
Biocatalysis and synthetic biology
Directed evolution is widely applied in more conventional contexts such as biocatalysis and synthetic biology (groundbreaking applications of biotechnological methods), especially in biotechnology where novel bioactive molecules and biomaterials can be produced. Artificial enzymes with new functions that did not previously occur in nature can be constructed by directed evolution.
Improving enzyme selectivity and specificity
Directed evolution is a powerful strategy to create enzymes with high stereoselectivity, regioselectivity and chemoselectivity. Experimental challenges and analytical costs upon screening huge populations of mutants can be overcome by this methodology. Stereoselective, regioselective and chemoselective enzymes can be generated by this methodology.
Enhancing current enzymes
the same strategy can be used to enhance the properties of existing enzymes and to endow proteins with new functions. Improving endogenous inulinase production by directed evolution.
Automated in vivo enzyme engineering
High-throughput screening of evolved enzyme activities in vivo with directed evolution in vitro to accelerate the discovery of biocatalysts.
Applications of genetically encoded biosensors
Through the combination of directed evolution with GEBs (genetically encoded biosensors) converting the concentration of small‐molecule ligands into a more easily detectable signal, authors have been quite successful at identifying new or improved enzyme variants.
Using these technologies, customers can create innovation and exploit breakthroughs at the interface of their fields of interest and those listed below: biocatalysis, synthetic biology, multiplexed directed evolution, refinement of existing enzyme properties (for example, improving existing enzyme selectivity and improving existing enzyme specificity) and, eventually, fully automated in vivo enzyme engineering.
As we want the customers of our Enzyme Directed Evolution Service to be better informed, here are some questions that are commonly asked, along with answers:
Q: What is enzyme-directed evolution technology?
A: Directed evolution of enzymes improves an enzymatic reaction by mimicking natural selection. The method can increase activity, stability and selectivity of the enzyme within a week of use.
Q: What is your company's enzyme directed evolution platform?
A: Our company uses the EnzymoGenius™ technology platform, with its own independent intellectual property rights. The platform organises the information in natural enzymes and can create original enzymes, and creates the expectation of the biomanufacture of key chemicals.
Q: What steps does your company's service process include?
A: Our company's service process covers the entire process from discovery of new enzymes, expression and purification to key technical steps for the rapid discovery of new enzymes.
Q: Can your company's services quickly discover new enzymes?
A: Yes, my company’s technology can find new enzymes. An enzyme directed evolution service can screen enzymes for your substrate with desired characteristics on short notice.
Q: How long does it take for directed evolution services?
A: How long the directed evolution takes depends on factors such as how many orders of magnitude of improvement in enzyme performance are needed to solve the problem, the size of the mutation pool, and, most importantly, how efficient the screening and selection steps are (which in turn is affected by the assay type). Either high-throughput screening or high-throughput selection, or both, offer ways of making screening and selection much more efficient.
Q: What is the cost of directed evolution services?
A: Average costs for directed evolution services start in the five-figure range per project, and are generally determined by the size of the experiment needed, the difficulty of screening, and the number of iterations needed. High-throughput technology and computer-aided design reduce costs.
Q: What is the success rate of directed evolution services?
A: The outcome is highly dependent on the starting properties of the enzyme and the goal of enhancement. However, through the use of advanced screening (such as multi-agent screening), and enhanced selection (such as continuous directed evolution), success rates can be substantially improved.
Q: Are your company's services suitable for all types of enzymes?
A: You are offering services geared toward optimising the functioning, stability and selectivity of enzymes that their customers might need. The enzyme types the process can be employed on are as per customer specification.
Q: Does your company's services provide technical support and follow-up services?
A: In addition to actually performing the rounds of enzyme directed evolution for your customer, your company will also provide technical support and follow-up services to enable your client to use your optimised enzymes smoothly.
Customers can better understand our Enzyme Directed Evolution Service and make more reasonable choice and use after to clear up the answers for the above questions and answers.
Our service includes screening the discovery of new enzymes for you to express and purify to obtain new enzymes quickly. We have a professional R & D team and more than 10 years of experience in R & D outsourcing service, and we provide customers with high quality service. In case of any requirements or questions, feel free to contact us.
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