Pseudomonas putida production

A very efficient and universal method has been developed for the production of optically pue L- and D-amino adds. The prindple is based on the enantioselective hydrolysis of D,L-amino add amides. The stable D,L-amino add amides are effidently prepared under mild reaction conditions starting from simple raw materials (Figure A8.2). Thus reaction of an aldehyde with hydrogen cyanide in ammonia (Strecker reaction) gives rise to the formation of the amino nitrile. The aminonitrile is converted in a high yield to the D,L-amino add amide under alkaline conditions in the presence of a catalytic amount of acetone. The resolution step is accomplished with permeabilised whole cells of Pseudomonas putida ATCC 12633. A nearly 100% stereoselectivity in hydrolysing only the L-amino add amide is combined with a very broad substrate spedfidty. 

Harayama S, M Rekik, M Wubbolts, K Rose, RA Leppik, KN Timmis (1989) Characterization of five genes in the upper-pathway operon of TOE plasmid pPWWO from Pseudomonas putida and identification of the gene products. J Bacterial 171 5048-5055. 

The production of carbon monoxide from trichloroflnoromethane catalyzed by cytochrome P450c nj proceeded through intermediate formation of the dichloroflnorocarbene (Li and Wackett 1993) (Fignre 7.70c). Other reactions included (3-elimination from l,l,l-trichloro-2,2,2-trifiuorethane (Fignre 7.70c). Pseudomonas putida strain G786 (pGH-2) was constructed to contain both the... 

Donnelly Ml, S Dagley (1980) Production of methanol from aromatic acids by Pseudomonas putida. J Bacterial 142 916-924. 

The oxidation of morphine by Pseudomonas putida MIO gave rise to a large number of transformation products including hydromorphone (dihy-dromorphinone), 14/3-hydroxymorphine, 14 6-hydroxymorphinone, and dihydromorphine. Similarly, in incubations with oxymorphone (14/3-hydroxy-... 

Scheme 2 Structures of the benzylisoquinoline alkaloid laudanosine and its transformed products (2, 3, and 4) by utilizing Pseudomonas putida incubated at 30 °C for 96 h [47]... 

The inifial sfeps in fhe mefabolism of morphine and codeine by Pseudomonas putida MIO involve oxidafion of fhe C-6 hydroxy group and subsequenf reducfion of fhe 7,8-olefinic bond, forming hydromorphone (dihydromorphinone) and hydrocodone (dihydrocodeinone), respectively (Scheme 4) [52], These products have important industrial appUcations hydromorphone is an analgesic some seven times more potent than morphine [53],... 

Wu, S., Fallon, R.D. and Payne, M.S. (1997) Over-production of stereoselective nitrile hydratase from Pseudomonas putida 5B in Pichia pastoris activity requires a novel downstream protein. Applied Microbiology and Biotechnology, 48 (6), 704—708. 

Other approaches could use transcriptional regulators that directly bind the substrate or product of the reaction and activate the reporter gene. For instance, a mutant transcriptional activator from Pseudomonas putida, NahH, was used that can bind various benzoic acids to develop a screening/selection method to detect the action of benzaldehyde dehydrogenase [45]. A transcriptional regulator may need to be engineered to bind the desired compound before it can be used in such a manner [46]. 

Martin, A. B. Alcon, A. Santos, V. E., and Garcia-Ochoa, F., Production of a Biocatalyst of Pseudomonas Putida CECT5279 for Dibenzothiophene (DBT) Biodesulfurization for Different Media Compositions. Energy Fuels, 2004. 18(3) pp. 851-857. 

Ben-Mansour H, Mosrati R, Corroler D et al (2009) In vitro mutagenicity of Acid Violet 7 and its degradation products by Pseudomonas putida mt-2 correlation with chemical structures. Env Toxicol Phamacol 27 231-236... 

The enzyme p-ethylphenol methylene hydroxylase (EPMH), which is very similar to PCMH, can also be obtained from a special Pseudomonas putida strain. This enzyme catalyzes the oxidation of p-alkylphenols with alkyl chains from C2 to C8 to the optically active p-hydroxybenzylic alcohols. We used this enzyme in the same way as PCMH for continuous electroenzymatie oxidation of p-ethylphenol in the electrochemical enzyme membrane reactor with PEG-ferrocene 3 (MW 20 000) as high molecular weight water soluble mediator. During a five day experiment using a 16 mM concentration of p-ethylphenol, we obtained a turnover of the starting material of more than 90% to yield the (f )-l-(4 -hydroxyphenyl)ethanol with 93% optical purity and 99% enantiomeric excess (glc at a j -CD-phase) (Figure 14). The (S)-enantiomer was obtained by electroenzymatie oxidation using PCMH as production enzyme. 

Aside from the multifaceted chemical conversions, there are sources to develop into industrially viable microbial conversions. 1,2,4-Butanetriol, for example, used as an intermediate chemical for alkyd resins and rocket fuels, is currently prepared commercially from malic acid by high-pressure hydrogenation or hydride reduction of its methyl ester. In a novel environmentally benign approach to this chemical, wood-derived D-xylose is microbially oxidized to D-xylonic acid, followed by a multistep conversion to the product effected by a biocatalyst specially engineered by inserting Pseudomonas putida plasmids into E. coli ... 

In an original application, Yasuda et al have used both l-AAO and d-AAO, and L-lysine oxidase to oxidize o ,Ci -diamino acids. The reactions produce the expected a-keto w-amino acid products, but these then spontaneously cyclize to form cyclic a-imino acids. These compounds are then substrates for the authors recently discovered A methyl amino acid dehydrogenase (NMAADH) from Pseudomonas putida, producing the pure L-cyclic amino acid (Scheme 5). 

Biological. Vinyl chloride was reported to be a biodegradation product from an anaerobic digester at a wastewater treatment facility (Howard, 1990). Under aerobic conditions. Pseudomonas putida oxidized 1,1,2-trichloroethane to chloroacetic and glyoxylic acids. Simultaneously, 1,1,2-trichloroethane is reduced to vinyl chloride exclusively (Castro and Belser,... 

Enhancement of metabolism of " C-toxaphene by several added cofactors was studied in 8 hrs aerobic cultures of washed Pseudomonas putida cells. The results of these experiments are summarized in Table 6. In the NAD group, NADH showed the greatest stimulatory effect on the production of total polar metabolites (1.2 times) over that of the 8 hrs incubate value. Similarly, FAD was responsible for a modest increase (1.1 times). The combination of NADH-FAD showed stimulation above that of either of the unpaired cofactors (2.0 times). Addition of mannitol to NADH-FAD stimulated this combination s total polar metabolite value from 2.0 to 2.3 times that of the 8 hrs incubate. In the sequence NADH to NADH-FAD to NADH-FAD-mannitol, the increase in the radioactivities in the ethyl acetate phase (moderately polar metabolites)... 

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