Towards the World of Custom-Made Biocatalysts - Enzymes by Design Enabling a Sustainable Bioindustrial Future
ID: 3436

Abstract: With demands on industry to create products more cost-efficiently, sustainably, safely, and with less waste, enzymes are becoming increasingly attractive solutions for carrying out chemical transformations. This panel showcases cutting-edge technology for enzyme development and recent advances in biocatalysis in order to deliver on this demand.

Christopher Savile (Codexis) will present the CodexTM biocatalyst panel, a set of enzyme variants that are designed to accept a wide range of substrates, produce different stereoisomers, be chemically robust, and be manufactured at commercial scale. The plate variants are also designed to yield structure-function information that can be used to provide a jumpstart to evolution. To achieve these goals, a development strategy was conceived consisting of four steps: evolve a robust backbone, generate binding pocket diversity, screen on multiple diverse substrates, and select variants with broad phenotypic diversity. This strategy was then used to produce five Codex panels: ketoreductase, transaminase, ene reductase, acylase, and halohydrin dehalogenase. This strategy as well as the development of two of these panels, the ketoreductase and ene reductase, will be presented.

Daniela Grabs (Arzeda) will describe the application of the Rosetta computational enzyme design methodology to construct new catalysts for reactions with no natural enzyme. Our computational methodology is general and applicable to any reaction. It starts with a description of the ideal active site - the transition state model surrounded by functional groups optimally oriented for catalysis. Next, protein scaffolds are identified which can accommodate the described active site. The remaining residues in the active site are then computationally redesigned to generate a shape and charge complementary binding pocket for high specificity and affinity. The Arzeda technology can transform biocatalysis with the ability to create enzymes on demand to expand the enzyme toolkits. Examples of designed enzymes with two novel activities and their experimental validation are presented.

Dr. Oliver May (DSM) will point out that the “design” criteria applied in nature can significantly deviate from the requirements of industrial applications. For example, high substrate specificity can be an important feature of a natural enzyme whereas broad substrate specificity is desirable for many industrial enzymes. Obviously, efficient methods that allow us to specifically tailor enzyme properties towards industrial relevant needs will therefore hugely expand the scope of industrial biocatalysis. In the presentation different examples of structure guided and evolutionary enzyme engineering will be discussed for following biocatalysts and applications:

1) Deoxyribose aldolase for production of statin intermediate

2) Hydroxynitrile lyases for production of agro and pharma intermediates

3) Amino acid amide racemase for production of aliphatic amino acids

4) Pen-G acylases for production of beta-lactam antibiotics

Current directed evolution strategies for building and screening libraries generally involve generating protein sequence diversity randomly across the whole sequence or in controlled random fashion at defined positions within the protein. These libraries generally have a large number of members that are "negative" with respect to the primary property, and require large numbers (10^4 to 10^6) of members be screened in order to find the relatively small number of positive mutations. As positive mutations accumulate for one property, all others get worse. David Estelle (Genencor) will present a method to rapidly create proteins with the desired combination of properties. This method uses a survey of sequence activity relationships for all properties as part of the design process and provides a rapid and efficient way to engineer proteins for multiple properties. Data generated this way for several proteins has given us insight into fundamental principles of protein sequence / activity relationships.













Moderator: Daniela Grabs, ARZEDA (United States)

Presenter 1:
Mike Martino, ARZEDA, (United States)  []

Presenter 2:  
Theo Sonke, DSM Pharmaceutical Products, (Netherlands)  [Confirmed]

Presenter 3:  
David Estell, Genencor® A Danisco Division, (United States)  [Confirmed]

Presenter 4 (if necessary): Development of the Codex Biocatalyst Panels  
Marissa Mock, Codexis, (United States)  [Confirmed]

Panel Organizer:
Daniela Grabs, ARZEDA, (United States)

Why should your submission should be selected for this year’s program?
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Having Genencor, DSM, Codexis and Arzeda together on one panel will bring together the world leader in enzyme engineering dedicated to showcase cutting-edge technology for enzyme development and recent advances in biocatalysis. And we are confident that the name recognition and the different spaces of the four companies will draw attention and interest from a broad audience.