With the support of our advanced protein engineering platform that we have established for many years, Creative BioMart has successfully designed a variety of platforms for different applications to produce special proteins based on the directed evolution strategy. Creative BioMart provides customers with a wide range of combinatorial enzyme engineering services, which can be used to broaden the range of enzyme substrates, increase stability in high temperature, acidic pH or organic solvents, and improve enantioselectivity.

Enzyme engineering is a process of improving the efficiency or activity of existing enzymes by changing the amino acid sequence of existing enzymes. This technology has been developed as a potential tool to overcome the shortcomings of natural enzymes as biocatalysts. Enzyme engineering has a wide range of applications in various industries such as baking, beverages, detergents, food, feed, leather, pharmaceuticals and textiles. For example, enzymes can degrade toxic compounds (amines, nitriles, phenols, and xenobiotic compounds) in industrial and domestic waste. Hydrolase or pectinase is added when preparing dairy products or fruit juices. Therefore, the research on enzyme engineering has aroused widespread interest.

Fig 1. Enzyme engineering strategies for the improvement of catalytic performance of industrial enzymes. (Sharma A, et al., 2021 )

Services

The current enzyme engineering strategies and methods are mainly to modify and optimize key catalytic performance, structural stability, and introduce new features to meet the growing needs of enzyme application processes. Creative BioMart has successfully established an enzyme engineering platform to customize existing enzymes and construct high-efficiency new industrial enzymes to produce value-added products cost-effectively. The enzyme engineering services we provide include but not limited to:

• Enzyme Stability Enhancement Service
• Protease Substrate Screening Service
• Enzyme Activity Analysis Service
• Protease Substrate Specificity Profiling Service

Understanding the relationship between enzyme stability and activity is critical to improving the function of enzymes. These characteristics must be considered when designing enzymes, including catalytic activity, substrate specificity, enantioselectivity, thermodynamic stability, co-solvent stability, expressibility and solubility. Our scientists are committed to developing a variety of methods to explore enzyme sequences and create new high-efficiency biocatalysts. Our enzyme engineering platform has achieved remarkable results in the new design and optimization of enzymes for chemical and pharmaceutical biosynthesis, regenerative medicine, food production, waste biodegradation and biosensing.

Methods of Enzyme Engineering

For your project, we will choose the right strategy for you. These technologies can be used alone or in combination to significantly improve the properties of the enzyme:

• Directed evolution: we use random mutation, error-prone PCR, DNA recombination and staggered expansion methods, and then high-throughput screening and selection of libraries to identify mutants with ideal characteristics. In addition, we combine directed evolution with metabolic engineering to obtain improved or completely new metabolic pathways, thereby developing new whole-cell biocatalysts for industrial applications.
• Saturation mutagenesis: we apply this technology to the hot spots of the enzyme, which can improve the catalytic efficiency and thermal stability of the enzyme.
• Fusion: the chimeric enzyme produced by fusion technology improves the catalytic efficiency, thermal stability, product selectivity and substrate specificity.
• Truncation: our truncation strategy is used to remove excess regions in the protein that hinder enzyme activity. Truncated enzyme libraries or enzymes can be constructed with altered enzyme properties.
• Rational design: the chimeric enzyme produced by fusion technology improves the catalytic efficiency, thermal stability, product selectivity and substrate specificity.
• Sense codon reassignment: in E. coli, incorporation of specific cotranslational phosophoserine into any targeted position in a protein can be carried out by using sense codon rearrangement technique.
• Computational tools for rational engineering: in order to improve enzyme activity, specificity and stability, we have developed several calculation tools and software. CAVER software has been used for the identification and analysis of tunnels and channels in protein structures. YASARA has been widely used in molecular mechanics-based simulation and graph-based hot spot detection as a reasonable enzyme engineering.
• Semi-rational design: it creates a smaller and smarter library based on biochemical or structural information, which has a high advantage.

Service Principle

Creative BioMart is based on innovative, pragmatic and honest, adhering to the tenet of "Quality is our life, providing customers with the best quality service", providing customized services to customers around the world.

We will be glad to discuss details of intended interaction studies with you and develop experimental strategies/methods tailored to your requirement. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, Monday to Friday. Please do not hesitate to contact us for more information or to discuss in detail, if you are interested in our services.

Reference

1. Sharma A, Gupta G, et al.. (2021) Enzyme engineering: current trends and future perspectives. Food Reviews International. 37(2): 121-154.

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