Pseudomonas immobilization

For the kinetic resolution of racemic a-acetoxyamide 6 several native enzymes were used (Scheme 5.5). The native Upases from Pseudomonas cepacia (PCL) and porcine pancreas (PPL) showed the highesL although stiU unsatisfactory, enantios-electivity ( = 5.1 and 3.5, respectively). Upon immobilization into a solgel matrix, the enantioselectivity of PCL was improved significantly to 30.5. The covalent immobilization on Eupergit increased the enantioselectivity even more (34.0) [23]. 

The low-temperature method was then applied to the resolution of ( )-2-hydroxy-2-(pentafluorophenyl)acetonitrile (7) (Fig. which is usahle for the syntheses of a variety of ethane diols, amino alcohols containing CgFj groups as novel chiral ligands. After screening lipases such as Amano PS and AK, lipase LIP Pseudomonas aeruginosa lipase immobilized on Hyflo Super-Cel, Toyobo,... 

Later, in a modification to the above system, we reported the use of an indenylruthenium complex 2 as a racemization catalyst for the DKR of secondary alcohols, which does not require ketones but a weak base hke triethylamine and molecular oxygen to be achvated. The DKR with 2 in combination with immobilized Pseudomonas cepacia lipase (PCL, trade name. Lipase PS-C ) was carried out at a lower temperature (60°C) and provided good yields and high optical purities (Table 2). This paved the way for the omission of ketones as... 

Immobilization of neutral xenobiotics in soils by qnatemary ammoninm cations has been established, and its significance on the bioavailability of naphthalene to bacteria has been examined. Bioavailability was determined by the rates of desorption, and these differed between a strain of Pseudomonas putida and one of Alcaligenes sp. (Crocker et al. 1995). 

Mordocco A, Jenkins CKR (1999) Continuous degradation of phenol at low concentration using immobilized Pseudomonas putida. Enzyme Microb Technol 25 530-536... 

DKR with lipase TL (from Pseudomonas stutzeri) was part of the reaction scheme in work reported by Hoyos et al. [24]. Here the lipase is immobilized using a number of... 

Furui, M. and Yamashita, K. (1983) Pressurized reaction method for continuous production of L-alanine by immobilized Pseudomonas dacunhae cells. Journal of Fermentation Technology, 61, 587—591. 

D-Aspartic acid 16 Apoxycillin Antibacterial Decarboxylation Pseudomonas dacunhae Immobilized cells [10]... 

In these inventions, R. rhodochrous IGTS8 ATCC 53968 [67] can be used, as well as a list of some other patented biocatalysts, such as Pseudomonas sp. CB1 ATCC No. 39381 [7], Acinetobacter sp. CB2 ATCC No. 53515 [8], B. Sulfasportare [16], B. sphaericus IGTS9 ATCC No. 53969 [91], the mixed cultures ATCC No. 39327 [159] and those described in Ref. [160], Valentine discloses the use of all possible biocatalyst types, namely cell cultures (free, immobilized or fragmented), cell extracts, enzyme mixtures (extracted or synthesized) or mixtures of all of them. 

In conventional synthetic transformations, enzymes are normally used in aqueous or organic solvent at moderate temperatures to preserve the activity of enzymes. Consequently, some of these reactions require longer reaction times. In view of the newer developments wherein enzymes can be immobilized on solid supports [183], they are amenable to relatively higher temperature reaction with adequate pH control. The application of MW irradiation has been explored with two enzyme systems namely Pseudomonas lipase dispersed in Hyflo Super Cell and commercially available SP 435 Novozym (Candida antarctica lipase grafted on an acrylic resin). 

Martin M, Mengs G, Plaza E, Garbi C, Sanchez M, Gibello A, Gutierrez F, Ferrer E (2000) Propachlor removal by Pseudomonas strain GCHII in an immobilized cell system. Appl Env Microbiol 66 1190-1194... 

Resmi et al. [59] used laterite stones for the immobilization of Pseudomonas putida (MTCC 1194). The amount of bacterial biomass attached to the support was 8.64 g/100 g of stones on dry weight basis. Packed bed reactor was used for treating mixture of seven azo dyes. With the help of immobilized bacterial strain, dye mixture was degraded to nontoxic smaller molecules. It was reported that even after 2 months, bacteria-coated pebbles were stable and suitable for the aerobic degradation of azo dyes. With the help of TLC and HPLC, 61.7% degradation was reported at the concentration of 50 pg/mL of dye. 

The bioconversion process of Acid Orange 7 will be hereby analyzed. This is an incremental study with respect to that due to Lodato et al. [41], based on the operation of an airlift reactor with cells of Pseudomonas sp. 0X1 immobilized on natural pumice (density = 1,000 kg/m3 particle size = 800-1,000 pm). Details regarding the strain, medium, culture growth and main diagnostics of the liquid phase are reported by Lodato et al. [41]. Elemental analysis of dry biomass was obtained by a C/H/N 600 LECO analyzer. 

Tests were carried out at 25°C and at initial pH 6.9. Cultures in the liquid medium were incubated in 50 mL Falcon tubes, continuously shaked at 220 rpm. Each culture contained a fresh Pseudomonas sp. 0X1 colony in 10 mL of medium. The airlift with 10 g of pumice was sterilized at 121°C for 30 min and then housed in a sterile room. One-day culture was transferred to the reactor and, after a batch phase, liquid medium with phenol as the only carbon source was continuously fed. The reactor volume V was fixed at 0.13 L. Aerobic conditions were established sparging technical air. Under these conditions microorganism started to grow immobilized on the solid s support. When immobilized biomass approached steady state, cyclic operation of the airlift was started by alternating aerobic/anaerobic conditions. 

Chang JS, Chou C, Chen SY (2001) Decolorization of azo dyes with immobilized Pseudomonas luteola. Process Biochem 36 757-763... 

For on-site measurement from grab sampling, a compact optical device with disposable strips for BOD determination has been developed [36]. The system includes three pairs of light-emitting diodes and photodiodes, and the disposable strips are made of inexpensive, transparent polycarbonate plates, where Pseudomonas fluorescens is immobilized. Using the 2,6-dichlorophenol-indo-phenol sodium salt as chromophore, a linear relationship was observed between the bioluminescence of the exposed strip and the BOD value. 

When cells of Pseudomonas sp. OS-K-29 immobilized on calcium alginate were incubated in 80 1 of a synthetic medium containing 0.2 vol.% of racemic 2,3-DCP, optically pure (S)-2,3-DCP of 100% e.e. was obtained. The immobilized cells could be re-used for a series of continuous reactions, a bioreactor being used for 19 reactions over 50 days without any loss of activity (Fig. 3). Optically pure (i )-2,3-DCP (100% e.e.) was also isolated from the racemate by means of stereospecific assimilation by Alcaligenes sp. DS-K-S38. Highly pure (R)- and (S)-EP of 99.5% e.e. were prepared from (S)- and (i )-2,3-DCP by treatment with aqueous NaOH, respectively [20-221 (Fig. 2). 

Similarly, lipase-catalyzed kinetic resolution has also been applied to intermediate nitrile alcohol 46 (Scheme 14.14). Best results were obtained by using immobilized Pseudomonas cepacia (PS-D) in diisopropyl ether, leading to excellent yield and enantiomeric excess of the desired (5)-alcohol 46a, along with (/J)-nitrile ester 47. Reduction of 46a with borane-dimethylsulhde complex, followed by conversion to the corresponding carbamate and subsequent lithium aluminum hydride reduction gave rise to the desired (S)-aminoalcohol intermediate 36, a known precursor of duloxetine (3). 

Chiral enantiopure 2,2 -dihydroxybiaryls are important as chiral ligands and are also structural motifs occurring in some natural products. Hydrolase-catalyzed resolution by acylation in organic solvents of some dihydroxybiaryls has been successfully achieved. Thus, the racemic binaphthols 111-113 have been resolved by mono acylation with vinyl acetate in t-butylmethyl ether ( ME) at 45 °C catalyzed by Pseudomonas sp. lipase (Scheme 4.35) [106]. In a similar way the 2,2 -dihydroxybiphenyl 114 can be acylated with vinyl acetate catalyzed by PSL immobilized on celite in TBME at 45 °C (Scheme 4.35) [107]. Butanolysis of the racemic monobutyrate of binaphthol rac-115 catalyzed by CALB in toluene at 80 °C for 72 h gives (R)-binaphthol (93% ) at ca. 50% conversion [108]. 

S)-N-(tert-butoxycarbonyl)-hydroxymethylpiperidine (8) is a key intermediate in the synthesis of a potent tryptase inhibitor (Scheme 7.5). It was synthesized from (R,.S)-3-hydroxyrnethylpiperidine via fractional crystallization of the corresponding L(-)dibenzoyl tartrate salt followed by hydrolysis and acylation [17]. The lipase from Pseudomonas cepacia (PS-30) immobilized on polypropylene accurel PP catalyzed the esterification of racemic 6 with succinic anhydride and toluene, giving the (S)-hemisuccinate ester (7). This was easily separated and hydrolyzed by base to the (S)-Boc-protected 3-hydroxymethylpiperidine (8). Using this repeated esterification procedure gave a 32% yield (maximum theoretical yield = 50%) and 98.9% . 

Havens and Rase reported the immobilization of an enzyme to degrade a specific organophosphate. The organophosphate was an agricultural grade material (parathion). The enzyme was harvested from recombinant Pseudomonas diminuta and immobilized by emulsifying a solution with a prepolymer. The product of the reaction was reported to have excellent stability and the method was proposed for cleanup of small spills of parathion. 

In addition, L-sorbose (25) has been converted into L-xty/o-2-hexulo-sonic acid (28) by way of L-xiy/o-hexos-2-ulose (44) by using mixtures of Gluconobacter melanogenus IFO 3293 and Pseudomonas syringae NRRL B-865 immobilized in a poly(acrylamide) gel.305 The yields were, however, low. 

Immobilized whole cells of Pseudomonas fluorescens were used for the glucose sensor. The microbial sensor has been applied to the continuous determination of glucose in molasses. 

I. Chibata, T. Tosa, and S. Takamatsu, Industrial production of L-alanine using immobilized Escherichia coli and Pseudomonas dacunhae, Microbiol. Sci. 1984, 1, 58-62. 

S. M. Cooper, J. E. Gavagan, B. Stieglitz, S. M. Hennessey, and R. DiCosimo, 5-Cyanovaleramide production using immobilized Pseudomonas chlororaphis B23, Bioorgan. Med. Chem. 1999, 7, 2239-2245. 

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