Proteases are widely used in the food industry for milk coagulation, meat tenderization, and flavoring, and in detergents as protein stain removal. With its advanced protein engineering platform established over the years, Creative BioMart has successfully developed engineered proteases with desirable pH and temperature activity and stability characteristics, allowing for more efficient functioning at low temperature and alkaline pH. Our scientists are committed to providing fully customized engineered proteases production solutions for food industry to customers around the world.

Introduction of Proteases Production

The main sources of proteases are animals, plants and microorganisms (bacteria and fungi). Proteases have a wide range of applications in the food industry, such as in brewing, meat tenderization, milk coagulation and as a digestive aid. In addition, proteases are used to improve the flavor, nutritional value, solubility, and digestibility of food proteins, as well as to alter their functional properties, including coagulation and emulsification. Proteases are widely used in the bakery industry for the production of bread, baked goods, etc. Proteases can also be used to accelerate cheese ripening, alter functional properties and reduce the allergenic properties of dairy products. Microbial protease production is considered to have the greatest biotechnological potential due to its low cost, high yield, and ease of genetic manipulation. In addition, microbial protease genes have also been investigated for enzymatic properties for protein engineering, elucidating the role of proteases in pathogenicity. At present, protein engineering technology has become an effective tool for improving protease. The study found that cold adaptation of mesophilic subtilisin-like proteases can be achieved through directed evolution. DNA recombination was also used to isolate new proteases with improved properties from the starting material of 26 subtilisins.

Fig 1. Plant proteases, their classifications, and level of investigation in meat studies. (Tantamacharik T, et al., 2018)

Solutions

Proteases are important to promote the rapid development of the food industry. Creative BioMart has successfully utilized a variety of protein engineering techniques to produce engineered proteases with specificity, thermostability and catalytic activity for the food industry. In addition, we produce this enzyme for use in the detergent industry. In order to provide customized high-quality proteases to global customers, we take the following solutions:

• Improving the stability of protease in medium containing organic solvent.
• Reducing the rate of competitive hydrolysis of acyl donor esters, especially when water is used as the reaction medium.
• Limiting the normally undesired proteolytic activity of proteases to prevent competitive peptide cleavage during synthesis reactions.
• Extending or altering the specificity or enantioselectivity of native enzymes.

Applications of Protein Engineering in Proteases Production

Protein engineering techniques enable the rational engineering of conventional proteases to better adapt them to process conditions through the study of the relationship between their properties and structures. We use technologies ranging from intentional chemical modifications to genetic modification of wild-type enzymes as support for protease production solutions.

• Chemical modification of protease

We have successfully chemically modified a variety of proteases through the active site. Here, enhancement of its esterolytic activity while minimizing amidolysis was achieved by chemically modifying the protease. Furthermore, they are made more stable and maintain their activity in organic solvent media.

• Site-direct and random mutagenesis generates mutations

We genetically modify protein sequences to increase the catalytic activity of proteases in organic reaction media. Here, not only the introduction of point mutations, but also the modification of entire protein domains can be achieved.

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

Reference

1. Tantamacharik T, Carne A, Agyei D, et al.. (2018) Use of plant proteolytic enzymes for meat processing[M]. Biotechnological Applications of Plant Proteolytic Enzymes. Springer, Cham. 43-67.

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