Lipases are widely used in the food industry to develop the flavor development of dairy products and the processing of other fatty foods. Creative BioMart has successfully developed lipases with substrate specificity, stereoselectivity and stability to improve the characteristic flavor of dairy products. Our scientists are committed to providing fully customized engineered lipases production solutions for food industry to customers around the world.

Introduction of Lipases Production

Fats and oils are important components of food. Lipases consist of a large family with the same overall structural fold, but with a multifunctional ring structure when in contact with a substrate and exhibiting a multifunctional substrate specificity. Lipases are used to produce flavor ex situ and modify structures by transesterification or transesterification to obtain products with increased nutritional value or suitable for parental feeding. Lipases are also used in foods to alter flavor by synthesizing short-chain fatty acid esters and alcohols. Currently, some classical strategies such as immobilization, additive or process engineering are used to produce lipase, but they cannot meet the requirements of a specific application, so its performance needs to be optimized. Rational protein design or directed (molecular) evolution has emerged as the most important approach to custom design lipases for specific applications. Scientists have successfully engineered triacylglycerol lipases and the Pseudomonas mendocina lipasemendocina with protein engineering.

Fig 1. Visualization of the target residues in the lipase from Rhizopus oryzae (ROL) for saturation mutagenesis to increase the stability of ROL towards aldehydes. (Bornscheuer U T, 2008)

Solutions

Lipases are important to promote the rapid development of the food industry. Creative BioMart has successfully utilized a variety of protein engineering techniques to produce fungi from the fungal family of rhizzomucor miehei and the bacterial family of Pseudomonas sp. and from Fusarium solani pisi lipase for the food industry. In addition, we produce this enzyme for use in the detergent industry and biofuels. We can develop new lipases for your specific application needs. In order to provide customized high-quality lipases to global customers, we take the following lipase protein engineering solutions:

• Variation addressing active sites.
• Variation addressing activity (including specific activity, substrate type specificity and chain length specificity).
• Variation addressing stability (including temperature stability, protease stability, and oxidative stability).
• Variant addressing cover function.
• Variation addressing macroscopic substrate interactions.
• The variant addresses calcium binding.
• Variants address detergent use and surfactant compatibility.
• Variants address X-ray structure and dynamics.

Applications of Protein Engineering in Lipases Production

Protein engineering is now widely used to alter the properties of lipid-modified lipases, including chemical, regio- and stereoselectivity, as well as long-term stability at high temperature or pH and activity in the presence of large substrate concentrations need to be improved. We use directed evolution combined with rational protein design to improve the performance of lipases:

• Rational design:

The key amino acid residues of the lipase are first determined from the protein structure (or homology model). These mutants were then introduced at the genetic level to generate and validate the desired properties of the lipase.

Our protein engineers have successfully used molecular modeling methods to identify molecular determinants of lipase acyl chain length specificity and to be able to significantly alter the chain length selectivity of lipases.

• Directed evolution:

Starting from one or a few (homologous) genes, using random mutagenesis methods from the resulting library of mutants or chimeras, high-throughput screening or selection methods are used to identify the desired variants for lipases.

Our protein engineers have successfully used specific genes to perform saturation mutagenesis at selected positions to rapidly and reliably identify stable mutants in libraries that improve the stereoselectivity and stability of lipases.

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

References

1. Bornscheuer U T. (2008) Alteration of lipase properties by protein engineering methods[J]. Oléagineux, Corps gras, Lipides. 15(3): 184-188.
2. Svendsen A. (2000) Lipase protein engineering[J]. Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular Enzymology. 1543(2): 223-238.

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