Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2-amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased.

Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2-amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased.

The present invention relates to the field of genetic engineering, particularly to method for improving thermo-stability of phytase, mutant and use. The present invention introduces a series of mutations to the phytase APPAmut4, which may involve introducing disulfide bonds, reducing the free energy of unfolding, optimizing the key residues in the coevolution process, and significantly improving the thermal stability of the phytase. Among the mutants of the present invention, the optimal mutant APPAmut9 retains about 70% of its activity after being treated for 5 minutes at 100 C., while the phytase APPAmut4 has already been inactivated. Therefore, the present invention overcomes the shortcomings of the prior art and provides phytase mutants with high thermal stability suitable for wide application in fields such as energy, food, and feed.

The present invention relates to the field of genetic engineering, particularly to method for improving thermo-stability of phytase, mutant and use. The present invention introduces a series of mutations to the phytase APPAmut4, which may involve introducing disulfide bonds, reducing the free energy of unfolding, optimizing the key residues in the coevolution process, and significantly improving the thermal stability of the phytase. Among the mutants of the present invention, the optimal mutant APPAmut9 retains about 70% of its activity after being treated for 5 minutes at 100 C., while the phytase APPAmut4 has already been inactivated. Therefore, the present invention overcomes the shortcomings of the prior art and provides phytase mutants with high thermal stability suitable for wide application in fields such as energy, food, and feed.

Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2-amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased.

Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2-amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased.

Compositions and methods for the production of L-glufosinate are provided. The method involves converting racemic glufosinate to the L-glufosinate enantiomer or converting PPO to L-glufosinate in an efficient manner. In particular, the method involves the specific amination of PPO to L-glufosinate, using L-glutamate, racemic glutamate, or another amine source as an amine donor. PPO can be obtained by the oxidative deamination of D-glufosinate to PRO (2-oxo-4-(hydroxy (methyl) phosphinoyl) butyric acid) or generated via chemical synthesis. PPO is then converted to L-glufosinate using a transaminase in the presence of an amine donor. When the amine donor donates an amine to PPO. L-glufosinate and a reaction by product are formed. Because the PPO remaining represents a yield loss of L-glufosinate, it is desirable to minimize the amount of PPO remaining in the reaction mixture. Degradation, other chemical modification, extraction, sequestration, binding, or other methods to reduce the effective concentration of the by-product. i.e., the corresponding alpha ketoacid or ketone to the chosen amine donor will shift the reaction equilibrium toward L-glufosinate, thereby reducing the amount of PPO and increasing the yield of L-glufosinate. Therefore, the methods described herein involve the conversion or elimination of the alpha ketoacid or ketone by-product to another product to shift the equilibrium towards L-glufosinate.

Bio-production processes that rely on biological activity can be very slow and inefficient, yet, the output of these processes may have significant value. Using biological enzymes outside their natural cellular environments offers a significant and largely untapped opportunity to enhance bio-production processes. By decoupling enzymes from their native contexts, one can modify both their sequences and structures in ways that are favorable for industrial applications. This system and method mixes substrate molecules, PPi, and enzymes that utilize PPi (PPi-dependent enzymes) in a reaction chamber and ends with stable-form products, including phosphorylated molecules.

Bio-production processes that rely on biological activity can be very slow and inefficient, yet, the output of these processes may have significant value. Using biological enzymes outside their natural cellular environments offers a significant and largely untapped opportunity to enhance bio-production processes. By decoupling enzymes from their native contexts, one can modify both their sequences and structures in ways that are favorable for industrial applications. This system and method mixes substrate molecules, PPi, and enzymes that utilize PPi (PPi-dependent enzymes) in a reaction chamber and ends with stable-form products, including phosphorylated molecules.