Optimization for a recombinantE. coli fed-batch fermentation

Applied Biochemistry and Biotechnology - The operating strategy that produces the maximum foreign protein expression for a fed-batch process is desired. This is achieved by using a feasible...     

Production of fumaric acid using rice bran and subsequent conversion to succinic acid through a two-step process

The fungal production of fumaric acid using rice bran and subsequent bacterial conversion of succinic acid using fungal culture broth were investigated. Since the rice bran contains abundant proteins, amino acids, vitamins, and minerals, it is suitable material that fungi use as a nitrogen source. The effective concentration of rice bran to produce fumaric acid was 5 g/L. A large amount of rice bran caused excessive fungal growth rather than enhance fumaric acid production. In addition, we could produce fumaric acid without the addition of zinc and iron. Fungal culture broth containing appro × 25 g/L of fumaric acid was directly employed for succinic acid conversion. The amount of glycerol and yeast extract required for succinic acid conversion was reduced to 70 and 30%, respectively, compared with the amounts cited in previous studies.     

Studies on nutrient requirements and cost-effective supplements for ethanol production by recombinantE. coli

This article describes a systematic study of the nutritional requirements of a patented recombinant ethanologenicEscherichia coli (11303:pLOI297) and provides cost-effective formulations that are compatible with the production of fuel ethanol in fermentations of lignocellulosic prehydrolysate characterized by high xylose conversion efficiency. A complex and nutrient-rich laboratory medium, Luria broth (LB), provided the benchmark with respect to fermentation performance standard. Xylose fermentation performance was assessed in terms of the target values for operational process parameters established by the US National Renewable Energy Laboratory (NREL)—final ethanol concentration (25 g/L), xylose-to-ethanol conversion efficiency (90%), and volumetric productivity (0.52 g/L·h). Biomass prehydrolysates that are rich in xylose also contain acetic acid, and in anticipation of a need to reduce acetic acid toxicity, the fermentors were operated with a pH control set-point of 7.0 Growth and fermentation in the minimal defined salts (DS) medium was only about 15% compared to the reference medium. Amendment of the minimal medium containing 6 wt% xylose with both vitamins and amino acids resulted in improved growth, but the volume productivity (0.59 g/L·h) was still only about 54% of that with LB (1.1 g/L·h). Formulations directed at cost reduction through the use of less expensive commercial complex nutritional supplements were within 90% of the NREL process target with respect to yield and provided a productivity at about 80% of the LB medium, but were not economical. Corn steep liquor (CSL) at about 7–8 g/L was shown to be a complete source of nutritional requirements and supported a fermentation performance approaching that of LB. At a cost of CSL of $50/t (dry wt), the economic impact of using this amount CSL as the sole nutritional supplement in a cellulosic ethanol plant was estimated to be about 4¢/gal of ethanol.

     

Production ofl-malic acid from fumaric acid by resting cells ofBrevibacterium sp.

Applied Biochemistry and Biotechnology - The ability of the resting cells ofBrevibacterium sp. B2D, a flocculent strain, to convert fumaric acid tol-malic acid was investigated under different...     

Chemistry of 2-bromo 3-trichloromethyl succinic anhydride and dimethyl ester : preparation of various halogenated succinic,maleic, fumaric and malic acid derivatives

Various reactions of the readily available adducts of bromotrichloromethane with maleic anhydride and dimethyl maleate and fumarate leading to halogenated derivatives of succinic, maleic, fumaric or malic acids and/or esters are described.     

Production of succinic acid by anaerobiospirillum succiniciproducens

The effect of an external supply of carbon dioxide and pH on the production of succinic acid byAnaerobiospirillum succiniciproducens was studied. In a rich medium containing yeast extract and peptone, when the external carbon dioxide supply was provided by a 1.5M Na₂CO₃ solution that also was used to maintain the pH at 6.0, no additional carbon dioxide supply was needed. In fact, sparging CO₂ gas into the fermenter at 0.025 L/L-min or higher rates resulted in significant decreases in both production rate and yield of succinate. Under the same conditions, the production of the main by-product acetate was not affected by sparging CO₂ gas into the fermenter. The optimum pH (pH 6.0) for the production of succinic acid was found to be in agreement with results previously reported in the literature. Succinic acid production also was studied in an industrial-type inexpensive medium in which light steep water was the only source of organic nutrients. At pH 6.0 and with a CO₂ gas sparge rate of 0.08 L/L-min, succinate concentration reached a maximum of 32 g/L in 27 h with a yield of 0.99 g succinate/g glucose consumed.     

Crystal structure of 2,2-dimethyl succinic acid.

The title compound crystallizes triclinically in space group of P1. The C2-COOH and C3-COOH molecular groups are planar. The crystal structure is stabilized by the formation of intermolecular (O-HO) hydrogen bonds.     

Experimental and quantum-chemical studies of the reactions of 6-methyluracil with succinic and fumaric acids

Possible structures of 6-methyluracil complexes with succinic and fumaric acids are studied by quantum-chemical means. The possibility of complex formation occurring between 6-methyluracil and the acids in the ionized and nonionized states is evaluated. The form of the complexes containing the nonionized acid is found to dominate. The quantum-chemical calculation data are consistent with the experimental results.     

Purification process for succinic acid produced by fermentation

Applied Biochemistry and Biotechnology - Succinic acid is a versatile four-carbon dicarboxylic acid. It can be used commerically as an intermediate chemical for the manufacture of 1,4-butanediol,...     

Bioconversion of municipal solid waste to lactic acid byLactobacillus species

Applied Biochemistry and Biotechnology - EightLactobacillus species were screened for production of lactic acid from acid-hydrolyzed municipal solid waste (AHMSW). Screening criteria included...     

Production of fumaric acid by immobilized rhizopus using rotary biofilm contactor

Rotary biofilm contactor (RBC) is a reactor consisting of plastic discs that act as supports for micro-organisms. The discs are mounted on a horizontal shaft and placed in a medium-containing vessel. During nitrogen-rich growth phase, mycelia ofRhizopus oryzae ATCC 20344 grew on and around the discs and formed the “biofilm” of self-immobilized cells on the surface of the plastic discs. During the fermentation phase, the discs are slowly rotated, and the biofilms are exposed to the medium and the air space, alternately. With RBC, in the presence of CaCO₃,Rhizopus biofilm consumes glucose and produces fumaric acid with a volumetric productivity of 3.78 g/L/h within 24 h. The volumetric productivity is about threefolds higher with RBC than with a stirred-tank fermenter with CaCO₃. Furthermore, the duration of fermentation is one-third of the stirred-tank system. The immobilized biofilm is active for over a 2-wk period with repetitive use without loss of activity.

     

Supramolecular organogels formed by monochain derivatives of succinic acid

The monoalkyl chain derivatives of succinic acid self-assemble into ordered bilayer aggregates by forming dimers of hydrogen bonded carboxylic acid in a number of organic solvents and finally gelatinize the solvents. The gelation ability of each derivative was inspected. The morphologies of xerogels were investigated by scanning electron microscope (SEM). The microstructure of aggregates was studied by small angle X-ray diffraction (SA-XRD) and Fourier transform infrared spectroscopy (FT-IR). The results reveal that the intermolecular hydrogen bonds between neighboring molecules are critical factor in the process of organogelation.     

Production of fumaric acid with immobilized biocatalysts

Immobilization ofRhizopus arrhizus mycelium improved fumaric acid production. The optimum conditions for fumaric acid production with immobilized cells were investigated using a statistical experimental design. Substrate concentration, carbon:nitrogen ratio, and residence time were chosen as independent variables. In the repeated batch shake flask fermentation, the fumaric acid yield from xylose was as much as 3.5 times higher with immobilized mycelium than with free mycelium. Polyurethane foam cubes, in this case, gave better results than nylon net cubes as a carrier.     

A novel fermentation pathway in anEscherichia coli mutant producing succinic acid,acetic acid,and ethanol

Escherichia coli strain NZN111, which is unable to grow fermentatively because of insertional inactivation of the genes encoding pyruvate: formate lyase and the fermentative lactate dehydrogenase, gave rise spontaneously to a chromosomal mutation that restored its ability to ferment glucose. The mutant strain, named AFP111, fermented glucose more slowly than did its wild-type ancestor, strain W1485, and generated a very different spectrum of products. AFP111 produced succinic acid, acetic acid, and ethanol in proportions of approx 2:1:1. Calculations of carbon and electron balances accounted fully for the observed products; 1 mol of glucose was converted to 1 mol of succinic acid and 0.5 mol each of acetic acid and ethanol. The data support the emergence in E.coli of a novel succinic acid:acetic acid:ethanol fermentation pathway.     

Metabolic modeling of fumaric acid production byrhizopus arrhizus

Applied Biochemistry and Biotechnology - A metabolic model is developed for fumaric acid production byRhizopus arrhizus. The model describes the reaction network and the extents of reaction in...     

Arylmaleic anhydrides via Heck arylation of fumaric acid

Palladium-catalyzed arylation of fumaric acid with aryl iodides is found to be a very simple, economic and scalable approach to arylmaleic anhydrides. The reaction is facilitated by the presence of donor moieties on the aryl fragment and does not occur with strong acceptor groups.     

Antifungal activity of fumaric acid in mice infected with Candida albicans.

An examination was made on the effect of fumaric acid on an experimental systemic candidiasis. Male ICR mice were innoculated into the tail veins with 10(6) yeast cells of Candida albicans and treated with daily intraperitoneal injections of fumaric acid at the dose of 40 mg/kg/d. The results indicated that the administration of fumaric acid was effective in prolonging the survival of animals after the fungal challenge and prevented one-fifth of the treated animals from dying of candidiasis.     

Determination of maleic acid and fumaric acid in the presence of each other by thermal analysis

The thermal properties of fumaric acid and maleic acid were investigated. It was found that differences in their thermal decompositions provide a possibility for their determination. The thermoanalytical determination of their mixtures was carried out derivatographically.     

Crystal engineering approach to forming cocrystals of amine hydrochlorides with organic acids. Molecular complexes of fluoxetine hydrochloride with benzoic, succinic, and fumaric acids

A crystal engineering strategy for designing cocrystals of pharmaceuticals is presented. The strategy increases the probability of discovering useful cocrystals and decreases the number of experiments that are needed by selecting API:guest combinations that have the greatest potential of forming energetically and structurally robust interactions. Our approach involves multicomponent cocrystallization of hydrochloride salts, wherein strong hydrogen bond donors are introduced to interact with chloride ions that are underutilized as hydrogen bond acceptors. The strategy is particularly effective in producing cocrystals of amine hydrochlorides with neutral organic acid guests. As an example of the approach, we report the discovery of three cocrystals containing fluoxetine hydrochloride (1), which is the active ingredient in the popular antidepressant Prozac. A 1:1 cocrystal was prepared with 1 and benzoic acid (2), while succinic acid and fumaric acid were each cocrystallized with 1 to provide 2:1 cocrystals of fluoxetine hydrochloride:succinic acid (3) and fluoxetine hydrochloride:fumaric acid (4). The presence of a guest molecule along with fluoxetine hydrochloride in the same crystal structure results in a solid phase with altered physical properties when compared to the known crystalline form of fluoxetine hydrochloride. On the basis of intrinsic dissolution rate experiments, cocrystals 2 and 4 dissolve more slowly than 1, and 3 dissolves more quickly than 1. Powder dissolution experiments demonstrated that the solid present at equilibrium corresponds to the cocrystal for 2 and 4, while 3 completely converted to 1 upon prolonged slurry in water.     

Electromigration of carrier-free radionuclide ions Bismuth complexes in aqueous solutions of oxalic, fumaric and succinic acids

The overall ion mobilities u of carrier-free radiobismuth have been measured in aqueous solutions of some dicarboxylic acids (H(2)L)-xalic, fumaric and succinic-by means of a new version of the electromigration method in electrolytes consisting of HClO(4)/H(2)L, 0.20m H(+), mu = 0.20m; Na(H)ClO(4)/H(2)L, 0.05m H(+), mu = 0.20m; Na(H)ClO(4)/H(2)L, 0.05m H(+), mu = 0.25m; at 298.15 K. Mathematical processing of the experimental functions u = f([L(2-)]) allowed calculation of the mean individual stability constants K(n) and ion mobilities u degrees of the complex ions [BiL(n)](3-2n), n = 1, 2: [Bi(C(2)O(4))](+), log K(1) = 7.65 (8), u degrees = +2.26 (5) x 10(-4) cm(2). sec(-1).V(-1); [Bi(C(2)O(4))(2)](-), log K(2) = 4.81 (2), u degrees = -1.63 (64) x 10(-4) cm(2).sec(-1).V(-1); [Bi(C(4)O(4)H(2))](+), log K(1) = 6.90 (20); [Bi(C(4)O(4)H(4))](+), log K(1) = 8.76 (48).