Enzymatic De-Racemization of Glufosinate for High-Purity L-Isomer Yield
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for producing chiral pure L-glufosinate ammonium, such as chiral resolution and chemical synthesis, are costly and inefficient, while existing biocatalysis methods face limitations in yield and raw material availability, making large-scale production challenging.
Innovation Solution
A glufosinate ammonium dehydrogenase mutant is used in a biological enzymatic de-racemization process, utilizing a multi-enzyme catalysis system to convert D,L-glufosinate ammonium into L-glufosinate ammonium, with a coenzyme cycling system to regenerate NAD(P)H, and a catalase to remove hydrogen peroxide byproducts.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If chiral resolution is used to separate D-type and L-type isomers, then optically pure L-glufosinate ammonium can be obtained, but the theoretical yield can only reach 50% and expensive chiral resolution reagents are needed
Solution Approach 1:
The patent changes the chemical parameter by using enzymatic catalysis instead of chemical resolution reagents. The L-glufosinate ammonium enolase catalyst selectively catalyzes the reaction of L-glutamate to form L-glufosinate ammonium, achieving both high optical purity and high theoretical yield by converting the racemic mixture directly to the desired enantiomer rather than separating it.
Solution Approach 2:
The patent replaces the mechanical separation process of chiral resolution with a biochemical catalytic process. Instead of using physical or chemical methods to separate enantiomers, the invention uses L-glufosinate ammonium enolase to selectively transform L-glutamate into L-glufosinate ammonium, achieving enantiomeric enrichment through selective catalysis rather than separation.
2Manufacturing precision
If chemical asymmetric synthesis is used from chiral raw materials, then optically pure L-glufosinate ammonium can be synthesized, but the process has many steps and low yield with high production cost
Solution Approach 1:
The patent extracts the key chiral information from L-glutamate and directly transfers it to form L-glufosinate ammonium through a single catalytic step. Instead of going through multiple synthesis steps from chiral raw materials, the invention directly converts the precursor to the target molecule with preserved chirality, achieving both high optical purity and production efficiency.
Solution Approach 2:
The patent uses L-glutamate as a pre-prepared chiral precursor that already contains the desired L-configuration. By using this pre-chiralized starting material and a single catalytic step, the invention avoids the need for multiple asymmetric synthesis steps, thereby improving both optical purity and production efficiency.
3Productivity
If direct enzymatic hydrolysis is used with L-glufosinate ammonium derivatives as substrates, then high conversion rate and ee value can be achieved, but expensive and difficultly available chiral raw materials are needed
Solution Approach 1:
The patent uses L-glutamate, which is a cheap and readily available amino acid, as the starting material instead of expensive and difficultly available chiral raw materials. The L-glufosinate ammonium enolase catalyst efficiently converts this inexpensive precursor into the desired product, achieving both high conversion rate and easy manufacturability.
4Ease of manufacture
If selective resolution of enzyme is used with precursor of racemic glufosinate ammonium, then raw materials are easy to obtain and catalyst activity is high, but the theoretical yield can only reach 50% resulting in waste of raw materials
Solution Approach 1:
The patent makes the useful action continuous by using the enzyme to catalyze the conversion of L-glutamate to L-glufosinate ammonium in a continuous manner. Instead of stopping at 50% yield from racemic resolution, the enzyme continuously processes the precursor to achieve high conversion rates, thereby eliminating raw material waste while maintaining ease of manufacture.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The process achieves high conversion rates and yields of L-glufosinate ammonium, reducing production costs and waste by using readily available D,L-glufosinate ammonium as a substrate without the need for expensive resolution reagents or additional separation steps.
Implementation Method 1
D-amino acid oxidase for catalyzing D-glufosinate ammonium in the D,L-glufosinate ammonium to 2-carbonyl-4-[hydroxy(methyl)phosphonyl]butanoic acid
Implementation Method 2
a glufosinate ammonium dehydrogenase mutant for catalytically reducing 2-carbonyl-4-[hydroxy(methyl)phosphonyl]butanoic acid to L-glufosinate ammonium
Implementation Method 3
a catalase to remove hydrogen peroxide byproducts
Data Source
AI summary
The present invention discloses a method for preparing L-glufosinate ammonium by biological enzymatic de-racemization, a glufosinate ammonium dehydrogenase mutant and a use thereof. The method for preparing L-glufosinate ammonium by biological enzymatic de-racemization includes catalyzing D,L-glufosinate ammonium as a raw material by a multi-enzyme catalysis system to obtain L-glufosinate ammonium. The enzyme catalysis system includes D-amino acid oxidase for catalyzing D-glufosinate ammonium in the D,L-glufosinate ammonium to 2-carbonyl-4-[hydroxy(methyl)phosphonyl]butanoic acid, and a glufosinate ammonium dehydrogenase mutant for catalytically reducing 2-carbonyl-4-[hydroxy(methyl)phosphonyl]butanoic acid to L-glufosinate ammonium. The glufosinate ammonium dehydrogenase mutant is obtained by mutation of glufosinate-ammonium dehydrogenase in wild fungi Thiopseudomonas denitrificans at a mutation site of V377S. The glufosinate ammonium dehydrogenase mutant in the present invention has better catalytic efficiency. When racemic D, L-glufosinate ammonium is used as a substrate for a catalytic reaction, the conversion rate is much higher than the conversion rate of a wild-type enzyme, and the yield of 2-carbonyl-4-[hydroxy(methyl)phosphonyl]butanoic acid (PPO for short) is also greatly improved.