Aldolases are regarded as an attractive route to produce biologically important compounds due to their ability to form carbon-carbon bonds. With its advanced protein engineering platform established over the years, Creative BioMart has successfully developed engineered aldolases with stability, substrate specificity and stereospecificity. Our scientists are committed to providing fully customized engineered aldolases production solutions for industry to customers around the world.

Introduction of Aldolases Production

The formation of carbon-carbon bonds is one of the key steps in the production of fine chemicals. Aldolase is one of the few enzymes that can catalyze the formation of carbon-carbon bonds. Some aldolase enzymes are involved in the degradation of metabolites (sugar degradation), others are used in the synthesis of the carbon backbone (amino acid synthesis). Aldolases convert their substrates to aldol products in high yields under mild conditions with high specificity. In most cases, these enzymes tightly control the relative and absolute configuration of newly formed neosteric centers. These properties make aldolase-catalyzed routes attractive for the production of biologically meaningful compounds. Therefore, aldolase enzymes are widely used in the biocatalytic synthesis of amino acids and carbohydrate derivatives. However, naturally occurring aldolase enzymes do not exist for many industrially important reactions and protein engineering, directed evolution and de novo enzyme design, which have all been used to alter stability, substrate specificity and stereoselectivity and other properties to produce tailored aldolase enzymes for use as biocatalysts.

Fig 1. Protein engineering of DHA-dependent aldolase. (Widmann M, et al., 2012)

Solutions

Creating an industrially applicable aldolase for biocatalysis plays an important role in promoting the green development of the industry. Creative BioMart has successfully utilized a variety of protein engineering techniques to improve the properties of aldolase enzymes, including their stability, substrate range and stereoselectivity. Our customers have extensively used aldolases for industrial biocatalytic synthesis of amino acids and carbohydrate derivatives. In order to provide customized high-quality aldolases to global customers, we take the following solutions:

• Enhancing aldolase activity and stability at lower temperatures.
• Broadening or altering the substrate specificity of aldehyde acceptors and ketone donors.
• Altering the stereochemistry of the reaction, the nucleophilic attack on the carbonyl carbon of the acceptor aldehyde occurs from the opposite side.

Applications of Protein Engineering in Aldolases Production

Scientists have increasingly used protein engineering to manipulate aldolase enzymes as new biocatalysts. Furthermore, the field of computational enzyme design has demonstrated the de novo design of aldolase to enhance their activity. We employed different strategies of rational protein engineering to improve the properties of aldolase and try to generate new aldolase.

• Key library design for directed evolution

Directed evolution has been shown to be an effective method to improve the properties of aldolase enzymes. In this way, our protein engineers designed highly concentrated libraries and resulted in mutants with increased selectivity or altered aldolase substrate specificity.

• Prediction of sequence-function relationships

Our protein engineers identify aldolase structurally or functionally relevant residues by analyzing multiple sequence alignments to identify relevant mutations, and predict mutants with improved substrate specificity, catalytic activity, or stability.

• Structure-based molecular modeling

Starting with protein structural information, our protein engineers seek to improve stability, activity, specificity or selectivity of aldolase through molecular modeling.

Creative BioMart has in-depth knowledge and experience of the tools and processes involved in engineered aldolases development, and provides professional aldolases production solutions for industry. Whether your aldolases are in the discovery and screening stages, or are planning engineered aldolases production, please contact us to discuss further details to ensure your next success.

Reference

1. Widmann M, Pleiss J, Samland A K. (2012) Computational tools for rational protein engineering of aldolases[J]. Computational and Structural Biotechnology Journal. 2(3): e201209016.

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