Amylases are widely used in the food industry to multiply functions for example it is used in the food industry for softening bread, adjusting flour for liquefaction and scarification of starch as well as juice treatment. Creative BioMart has successfully developed various α-amylases with stability, activity in the acidic range, and Ca²⁺-independence to improve food texture and shelf life. Our scientists are committed to providing fully customized engineered amylases production solutions for food industry to customers around the world.

Introduction of Amylases Production

Amylase, one of the main enzymes used in industry, hydrolyzes starch into polymers composed of glucose units. Most amylases and related enzymes belong to the α-amylase family, which contain a characteristic catalytic (beta/alpha)8-barrel domain. Amylases are extensively employed in the processed-food industry such as baking, brewing, preparation of digestive aids, production of cakes, fruit juices and starch syrups. α-amylases can be obtained from plants, animals and microorganisms. Among them, the α-amylases produced by bacillus licheniformis, bacillus stearothermophilus and bacillus amyloliquefaciens are dominant in industrial applications. Current studies have found that α-amylase is produced by submerged fermentation (SmF) and solid state fermentation (SSF), but it is difficult to control the matching properties for different applications. The development of protein engineering techniques can improve the properties of α-amylases, such as thermostability, pH profile, pH stability and Ca independence. Scientists have successfully developed calcium- and acid-independent starch-liquefying α-amylases using protein engineering techniques.

Fig 1. Applications of α-amylases. (Mehta D, et al., 2016)

Solutions

α-amylases are important to promote the rapid development of the food industry. Directed evolution and site-directed mutagenesis are powerful tools for engineering enzymes to improve their function and alter their properties, such as activity, selectivity, substrate specificity, stability, and solubility. Creative BioMart has successfully utilized a variety of protein engineering techniques to produce engineered α-amylases with stability, activity in the acidic range, and Ca independence for the food industry. We can develop new α-amylases for your specific application needs. Our process is:

(1) Based on the functional information and known three-dimensional structure of alpha-amylase, the targeted enzymes were subjected to site-directed mutagenesis.

(2) Introduction of random mutagenesis in the development of starch liquefaction alpha-amylase enzymes.

(3) After microbial screening, amylase variants with improved alkaline stability and calcium dependence were obtained.

Applications of Protein Engineering in Amylases Production

Protein engineering techniques can significantly improve α-amylases properties. In order to provide customized high-quality α-amylases to global customers, we take the following solutions:

• Increasing the thermostability of α-amylases:

We use methods such as site-directed mutagenesis, directed evolution, deletion mutagenesis to modify α-amylases to increase their thermostability.

• Altering the pH optimum and stability of α-amylases:

We employed site-directed mutagenesis to alter the enzyme's pH optimum and increase the enzyme's acid/base stability. This approach reduces the time and cost of screening libraries for improved variants.

• Improving oxidative stability of α-amylases:

We truncate the α-amylase at the N- and C-termini and further mutated it to increase thermostability and oxidative stability.

• Altering the transglycosylation/hydrolysis ratio of α-amylases:

We mutated maltogenic amylases to reduce the hydrolytic activity of the enzyme using a structure-directed protein engineering approach.

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

Reference

1. Mehta D, Satyanarayana T. (2016) Bacterial and Archaeal α-Amylases: Diversity and Amelioration of the Desirable Characteristics for Industrial Applications. Front Microbiol. 7:1129.

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