Cargill Inc.

Christopher Solheid

David B Weiner

Paula M Hicks

Sara C McFarlan

Steven John Gort

David Paul Weiner

Peter Luginbuhl

Fernando A Sanchez-Riera

Toby Howard Richardson

Lishan Zhao

Ellen Burke

Sherry R Kollmann

Brian J Brazeau

Jose Miguel Laplaza

Mervyn L De Souza

Wei Niu

Erin Kathleen Marasco

Analia Bueno

Joslin M Cuenca

Anil Bhagwan Khare

Mervyn Desouza

Joshua Lundorff

Brent Howard Hilbert

Kent Floy

Sara C Mcfarlan

Sherry R Kollman

Sara C Mc Farlan

This invention relates to polypeptides having aldolase activity, including pyruvate activity such as, without limitation, HMG and/or KHG aldolase activity, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. In some embodiments, the invention is directed to polypeptides having aldolase activity, including pyruvate activity such as, without limitation, HMG and/or KHG aldolase activity, including thermostable and thermotolerant activity, and polynucleotides encoding these enzymes, and making and using these polynucleotides and polypeptides. The polypeptides in accordance with the invention can be used in a variety of pharmaceutical, agricultural and industrial contexts. In some embodiments, the invention provides polypeptides and biosynthetic pathways that are useful in the production of R-2-hydroxy 2-(indol-3ylmethyl)-4-keto glutaric acid (R-MP) and certain stereoisomers of monatin, such as R,R and S,R monatin, and salts thereof, as well as certain stereoisomers of monatin derivatives, such as the R,R and S,R configurations, and salts thereof.

Aldolases, nucleic acids encoding them and methods for making and using them

Monatin and certain stereoisomers of monatin, such as R,R monatin and S,R monatin, as well as salts thereof, are produced using polypeptides and biosynthetic pathways. These polypeptides and biosynthetic pathways are also useful in the production of R-2-hydroxy-2-(indoly-3-ylmethyl)-4-keto glutaric acid, an intermediate that is formed in certain monatin synthesis pathways, including some biosynthetic pathways.

Polypeptides and biosynthetic pathways for the production of stereoisomers of monatin and their precursors

The invention provides for aminotransferase and oxidoreductase polypeptides and nucleic acids encoding such polypeptides. Also provided are methods of using such aminotransferase and oxidoreductase nucleic acids and polypeptides.

Aminotransferase and oxidoreductase nucleic acids and polypeptides and methods of using

Methods and systems for increasing the production of equilibrium reactions are described. In some embodiments, a method comprises removing enzymes from a reaction mixture once equilibrium is achieved, selectively re-adding enzymes and reactants for driving the product-forming reaction to product, and optionally recycling stabilized intermediates, by-products and/or co-products back into the reaction mixture. In some embodiments, a method comprises purifying the desired product from a reaction mixture and selectively recycling one or more components of the reaction mixture, such as original reactants, intermediates, co-products or by-products back into the reaction mixture as desired to improve titer. Systems for implementing the methods are also provided. In some embodiments the methods and systems are implemented for increasing the production of monatin and monatin derivatives, produced in multi-step equilibrium pathways.

Methods and systems for increasing production of equilibrium reactions

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Minneapolis, MN, United States of America

Christopher Solheid