53369-07-6

Usage

Hazard

Moderately toxic by ingestion

Upstream product

• 74-90-8 hydrogen cyanide 
• 59374-45-7 (3,3-Dimethoxy-propyl)-methyl-phosphinic acid ethyl ester 
• 15715-41-0 methyldiethoxyphosphine 
• 73385-92-9 ethyl methyl-(3,3-diethoxypropyl)phosphinate 
• 151-50-8 potassium cyanide 
• 36120-72-6 methyl-vinyl-phosphinic acid 2-chloro-ethyl ester 
• 1068-90-2 2-acetylaminomalonic acid diethyl ester 
• 40682-54-0 ethyl N-benzylideneglycinate 
• 63314-88-5 vinylmethylphosphinic acid methyl ester 
• 139574-94-0 methyl 2-(benzylideneamino)-4-(methoxymethylphosphinyl)butyrate 
• 87647-63-0 4-(Methoxy-methyl-phosphinoyl)-2-{[1-phenyl-meth-(E)-ylidene]-amino}-butyric acid ethyl ester 
• 71680-52-9 ammonium cyanide 
• 773837-37-9 sodium cyanide 
• 59374-49-1 ethyl methyl(3-propaldehyde)phosphinate 

Downstream Products

• 35597-44-5 L-phosphinothricin 
• 82785-28-2 4-[hydroxy(methyl)phosphonyl]-DL-homoalanine 

Relevant academic research and scientific papers

Tuning amino acid dehydrogenases with featured sequences for L-phosphinothricin synthesis by reductive amination

Biosynthesizing unnatural chiral amino acids is challenging due to the limited reductive amination activity of amino acid dehydrogenase (AADH). Here, for the asymmetric synthesis of L-phosphinothricin from 2-oxo-4-[(hydroxy)(methyl)phosphinoyl]butyric acid (PPO), a glutamate dehydrogenase gene (named GluDH3) from Pseudomonas monteilii was selected, cloned and expressed in Escherichia coli (E. coli). To boost its activity, a “two-step”-based computational approach was developed and applied to select the potential beneficial amino acid positions on GluDH3. L-phosphinothricin was synthesized by GluDH-catalyzed asymmetric amination using the D-glucose dehydrogenase from Exiguobacterium sibiricum (EsGDH) for NADPH regeneration. Using lyophilized E. coli cells that co-expressed GluDH3_V375S and EsGDH, up to 89.04 g L?1 PPO loading was completely converted to L-phosphinothricin within 30 min at 35 °C with a space-time yield of up to 4.752 kg·L?1·d?1. The beneficial substitution V375S with increased polar interactions between K90, T193, and substrate PPO exhibited 168.2-fold improved catalytic efficiency (kcat/KM) and 344.8-fold enhanced specific activity. After the introduction of serine residues into other GluDHs at specific positions, forty engineered GluDHs exhibited the catalytic functions of “glufosinate dehydrogenase” towards PPO.

L-GLUFOSINATE INTERMEDIATE AND L-GLUFOSINATE PREPARATION METHOD

Provided are L-glufosinate intermediate preparation method or L-glufosinate preparation method, the method, for preparing L-glufosinate intermediate or L-glufosinate from an L-homoserine derivative, comprising a step of preparing a compound of Chemical Formula 2 from a compound of Chemical Formula 1.

A Single-Transaminase-Catalyzed Biocatalytic Cascade for Efficient Asymmetric Synthesis of l-Phosphinothricin

A single-transaminase-catalyzed biocatalytic cascade was developed by employing the desired biocatalyst, ATA-117-Rd11, that showed high activity toward 2-oxo-4-[(hydroxy)(methyl)phosphinoyl] butyric acid (PPO) and α-ketoglutarate, and low activity against pyruvate. The cascade successfully promotes a highly asymmetric amination reaction for the synthesis of l-phosphinothricin (l-PPT) with high conversion (>95 %) and>99 % ee. In a scale-up experiment, using 10 kg pre-frozen E. coli cells harboring ATA-117-Rd11 as catalyst, 80 kg PPO was converted to ≈70 kg l-PPT after 24 hours with a high ee value (>99 %).

Preparation method of glufosinate-ammonium

The invention relates to a preparation method of glufosinate-ammonium (I). The method comprises the step of reacting an enantiomerically pure compound of formula (II) with a compound of formula (III) in the presence of a Lewis acid, wherein Hal is a halogen; PG is hydrogen or an amino protecting group; Z is OX or OY; R1 is a C1-C16 alkyl group, cyclohexyl group, cyclopentyl group or phenyl group, and each group can be substituted by hydrogen, a C1-C6 alkyl group, a C1-C6 alkoxy group or a dialkylamino group; R2 is a C1-C8 alkyl group, a C1-C8 ether group or a phenyl group; X and Y are respectively and independently alkyl, alkenyl or aryl; and chiral carbon atoms are marked with *. According to the method disclosed by the invention, high-purity glufosinate-ammonium can be obtained with high yield.

Development of a biocatalytic cascade for synthesis of 2-oxo-4-(hydroxymethylphosphinyl) butyric acid in one pot

2-Oxo-4-(hydroxymethylphosphinyl) butyric acid (PPO) is an important precursor compound for the broad-spectrum herbicide l-glufosinate (L-PPT). In this study, the gene of d-amino acid oxidase (DAAO) was cloned and expressed in Escherichia coli. By coupling exogenous catalase (CAT), a biocatalytic cascade was constructed for synthesis of PPO in one pot. The bioprocess was optimized on a 300 mL scale reaction by one factor at a time optimization. The conversion of this biocatalytic cascade achieved 46.8% towards 400 mM DL-PPT within 4 h. These results indicated that DAAO could be applied to the large-scale bioproduction of PPO and provide a promising route for the asymmetric synthesis of L-PPT by bio-enzymatic methods using PPO as the substrate.

Method for preparing L-glufosinate-ammonium

The invention belongs to the field of organic synthesis, and particularly relates to a method for preparing L-glufosinate-ammonium (I) or salt thereof. A compound represented by formula (II) or a saltthereof reacts with a compound represented by formula (III), and no matter whether an intermediate is separated or not, a product obtained by the reaction is subjected to a hydrolysis reaction to obtain L-glufosinate-ammonium (I) or a salt thereof. Compared with an existing L-glufosinate-ammonium synthesis route, the method is a new chemical synthesis route, the steps are simple, the atom economyis high, an L-glufosinate-ammonium product with a high ee value can be obtained without chiral catalysis, and the method has a potential industrialization application value.

Biocatalytic asymmetric synthesis of L-phosphinothricin using a one-pot three enzyme system and a continuous substrate fed-batch strategy

Transamination catalyzed by an aminotransferase is becoming a key tool for the production of chiral amine pharmaceuticals and agrochemicals owing to its excellent enantioselectivity and green credentials. To overcome the unfavorable thermodynamic equilibrium and the inhibition by the byproduct α-ketoglutaric acid in the transamination of 2-oxo-4-[(hydroxy)(methyl)phosphinoyl]butyric acid (PPO) to form the promising herbicide L-phosphinothricin (L-PPT), a tri-enzymatic cascade reaction system was developed by combining a robust glutamate dehydrogenase to recycle the byproduct to the amino donor L-glutamate in situ, together with a cofactor recycling process catalyzed by an alcohol dehydrogenase. Moreover, a continuous substrate fed-batch strategy was employed to alleviate the decomposition of PPO and applied to scale-up the cascade reaction to 90 L, yielding 111.4 g/L (615.4 mM) L-PPT in 99.7% yield and >99.9% ee with an productivity of 15.9 g/L?h. This combination of improved biocatalyst system and process engineering should prove to be economically competitive for industrial applications.

Semi-rational hinge engineering: modulating the conformational transformation of glutamate dehydrogenase for enhanced reductive amination activity towards non-natural substrates

The active site is the common hotspot for rational and semi-rational enzyme activity engineering. However, the active site represents only a small portion of the whole enzyme. Identifying more hotspots other than the active site for enzyme activity engineering should aid in the development of biocatalysts with better catalytic performance. Glutamate dehydrogenases (GluDHs) are promising and environmentally benign biocatalysts for the synthesis of valuable chirall-amino acids by asymmetric reductive amination of α-keto acids. GluDHs contain an inter-domain hinge structure that facilitates dynamic reorientations of the domains relative to each other. Such hinge-bending conformational motions of GluDHs play an important role in regulating the catalytic activity. Thus, the hinge region represents a potential hotspot for catalytic activity engineering for GluDHs. Herein, we report semi-rational activity engineering of GluDHs with the hinge region as the hotspot. Mutants exhibiting significantly improved catalytic activity toward several non-natural substrates were identified and the highest activity increase reached 104-fold. Molecular dynamics simulations revealed that enhanced catalytic activity may arise from improving the open/closed conformational transformation efficiency of the protein with hinge engineering. In the batch production of three valuablel-amino acids, the mutants exhibited significantly improved catalytic efficiency, highlighting their industrial potential. Moreover, the catalytic activity of several active site tailored GluDHs was also increased by hinge engineering, indicating that hinge and active site engineering are compatible. The results show that the hinge region is a promising hotspot for activity engineering of GluDHs and provides a potent alternative for developing high-performance biocatalysts toward chirall-amino acid production.

METHODS FOR IMPROVING YIELDS OF L-GLUFOSINATE

Compositions and methods for the production of L-glufosinate are provided. The method involves converting racemic glufosinate to the L-glufosinate enantiomer or converting PRO to L-glufosinate in an efficient manner. In particular, the method involves the specific amination of PRO to L-glufosinate, using L-glutamate, racemic glutamate, or another amine source as an amine donor. PRO 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. PRO is then converted to L-glufosinate using a transaminase in the presence of an amine donor. When the amine donor donates an amine to PRO, L-glufosinate and a reaction by product are formed. Because the PRO remaining represents a yield loss of L-glufosinate, it is desirable to minimize the amount of PRO 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 PRO 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.

Preparation method of glufosinate-ammonium

The invention discloses a preparation method of glufosinate-ammonium. The preparation method comprises the following steps that 1, in alkaline environment, 4-(hydroxymethyl phosphono)-2-carbonyl butyric acid (I) and a benzylamine solution react to produce 2-[( phenyl amino)-4-(methyl sodium phosphate)-sodium butyrate (II); 2, 2-[(phenyl amino)-4-(methyl sodium phosphate)-sodium butyrate (II) is subjected to acid hydrolysis to obtain glufosinate-ammonium (III). Compared with the prior art, the preparation method has the advantages that the conditions are mild; the yield of the glufosinate-ammonium is high; the purity is high.