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Enzymatic Production (enzymatic + production)
Selected AbstractsEnzymatic Production of l -Menthol by a High Substrate Concentration Tolerable Esterase from Newly Isolated Bacillus subtilis ECU0554ADVANCED SYNTHESIS & CATALYSIS (PREVIOUSLY: JOURNAL FUER PRAKTISCHE CHEMIE), Issue 3 2009Gao-Wei Zheng Abstract Enzymatic preparation of l -menthol has been attracting much attention in the flavor and fragrance industry. A new ideal strain, Bacillus subtilis ECU0554, which exhibited high hydrolytic activity and excellent enantioselectivity towards l -menthyl ester, has been successfully isolated from soil samples through enrichment culture and identified as Bacillus subtilis by 16S rDNA gene sequencing. The esterase extracted from B. subtilis ECU0554 (BSE) showed the best catalytic properties (E>200) for dl -menthyl acetate among the five menthyl esters examined. Enantioselective hydrolysis of 100,mM dl -menthyl acetate at 30°C and pH,7.0, using crude BSE as biocatalyst and 10% ethanol (v/v) as cosolvent, resulted in 49.0% conversion (3,h) and 98.0% ee for the l -menthol produced, which were much better than those using commercial enzymes tested. Moreover, BSE exhibited strong tolerance against high substrate concentration (up to 500,mM), and the concentration of l -menthol produced could reach as high as 182,mM, and more importantly, the optical purity of l -menthol produced was kept above 97% ee, which were not found in previous reports. These results imply that BSE is a potentially promising biocatalyst for the large-scale enzymatic preparation of l -menthol. Using this excellent biocatalyst, the enzymatic production of l -menthol will become a mild, efficient, inexpensive and easy-to-use "green chemistry" methodology. [source] Production of biodiesel: possibilities and challengesBIOFUELS, BIOPRODUCTS AND BIOREFINING, Issue 1 2007Sulaiman Al-Zuhair Abstract Biodiesel, defined as monoalkyl fatty acid ester (preferentially methyl and ethyl esters), represents a promising alternative fuel for use in compression-ignition (diesel) engines. Biodiesel fuel comes from renewable sources as it is plant- not petroleum-derived and as such it is biodegradable and less toxic. In addition, relative to conventional diesel, its combustion products have reduced levels of particulates, carbon oxides, sulphur oxides and, under some conditions, nitrogen oxides. Enzymatic production of biodiesel has been proposed to overcome the drawbacks of the conventional chemically catalyzed processes. The main obstacle facing full exploitation of the enzyme, lipase, potential is its cost. Therefore, reuse of lipase is essential from the economic point of view, which can be achieved by using the lipase in immobilized form. In addition, immobilized lipase displays improved stability and activity. Common immobilization techniques include attachment to solid supports and entrapment within the matrix of a polymer. This article presents a comparison between conventional processes and enzymatic processes and different possible feedstocks for biodiesel production. In addition, possible ways to overcome the problems facing the use of lipase are described. © 2007 Society of Chemical Industry and John Wiley & Sons, Ltd [source] Enzymatic production of ,- D -glucose-1-phosphate from trehaloseBIOTECHNOLOGY JOURNAL, Issue 9 2010Jef Van der Borght Abstract ,- D -Glucose-1-phosphate (,Glc1P) is an efficient glucosyl donor for both enzymatic and chemical glycosylation reactions but is currently very costly and not available in large amounts. This article provides an efficient production method of ,Glc1P from trehalose and phosphate using the thermostable trehalose phosphorylase from Thermoanaerobacter brockii. At the process temperature of 60°C, Escherichia coli expression host cells are lysed and cell treatment prior to the reaction is, therefore, not required. In this way, the theoretical maximum yield of 26% could be easily achieved. Two different purification strategies have been compared, anion exchange chromatography or carbohydrate removal by treatment with trehalase and yeast, followed by chemical phosphate precipitation. In a next step, ,Glc1P was precipitated with ethanol but this did not induce crystallization, in contrast to what is observed with other glycosylphosphates. After conversion of the product to its cyclohexylammonium salt, however, crystals could be readily obtained. Although both purification methods were quantitative (>99% recovery), a large amount of product (50%) was lost during crystallization. Nevertheless, a production process for crystalline ,Glc1P is now available from the cheap substrates trehalose and inorganic phosphate. [source] Enzymatic Production of l -Menthol by a High Substrate Concentration Tolerable Esterase from Newly Isolated Bacillus subtilis ECU0554ADVANCED SYNTHESIS & CATALYSIS (PREVIOUSLY: JOURNAL FUER PRAKTISCHE CHEMIE), Issue 3 2009Gao-Wei Zheng Abstract Enzymatic preparation of l -menthol has been attracting much attention in the flavor and fragrance industry. A new ideal strain, Bacillus subtilis ECU0554, which exhibited high hydrolytic activity and excellent enantioselectivity towards l -menthyl ester, has been successfully isolated from soil samples through enrichment culture and identified as Bacillus subtilis by 16S rDNA gene sequencing. The esterase extracted from B. subtilis ECU0554 (BSE) showed the best catalytic properties (E>200) for dl -menthyl acetate among the five menthyl esters examined. Enantioselective hydrolysis of 100,mM dl -menthyl acetate at 30°C and pH,7.0, using crude BSE as biocatalyst and 10% ethanol (v/v) as cosolvent, resulted in 49.0% conversion (3,h) and 98.0% ee for the l -menthol produced, which were much better than those using commercial enzymes tested. Moreover, BSE exhibited strong tolerance against high substrate concentration (up to 500,mM), and the concentration of l -menthol produced could reach as high as 182,mM, and more importantly, the optical purity of l -menthol produced was kept above 97% ee, which were not found in previous reports. These results imply that BSE is a potentially promising biocatalyst for the large-scale enzymatic preparation of l -menthol. Using this excellent biocatalyst, the enzymatic production of l -menthol will become a mild, efficient, inexpensive and easy-to-use "green chemistry" methodology. [source] A novel transverse push,pull microprobe: in vitro characterization and in vivo demonstration of the enzymatic production of adenosine in the spinal cord dorsal hornJOURNAL OF NEUROCHEMISTRY, Issue 1 2001Shawnna L. Patterson Adenosine produces analgesia in the spinal cord and can be formed extracellularly through enzymatic conversion of adenine nucleotides. A transverse push,pull microprobe was developed and characterized to sample extracellular adenosine concentrations of the dorsal horn of the rat spinal cord. Samples collected via this sampling technique reveal that AMP is converted to adenosine in the dorsal horn. This conversion is decreased by the ecto-5,-nucleotidase inhibitor, ,,,-methylene ADP. Related behavioral studies demonstrate that AMP administered directly to the spinal cord can reverse the secondary mechanical hyperalgesia characteristic of the intradermal capsaicin model of inflammatory pain. The specific adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine (CPT) inhibits the antihyperalgesia produced by AMP. This research introduces a novel microprobe that can be used as an adjunct sampling technique to microdialysis and push,pull cannulas. Furthermore, we conclude that AMP is converted to adenosine in the dorsal horn of the spinal cord by ecto-5,-nucleotidase and subsequently may be one source of adenosine, acting through adenosine A1 receptors in the dorsal horn of the spinal cord, which produce antihyperalgesia. [source] Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin CPROTEIN SCIENCE, Issue 1 2002Karin Fritz-Wolf Abstract Glutarylamidase is an important enzyme employed in the commercial production of 7-aminocephalosporanic acid, a starting compound in the synthesis of cephalosporin antibiotics. 7-aminocephalosporanic acid is obtained from cephalosporin C, a natural antibiotic, either chemically or by a two-step enzymatic process utilizing the enzymes D-amino acid oxidase and glutarylamidase. We have investigated possibilities for redesigning glutarylamidase for the production of 7-aminocephalosporanic acid from cephalosporin C in a single enzymatic step. These studies are based on the structures of glutarylamidase, which we have solved with bound phosphate and ethylene glycol to 2.5 Å resolution and with bound glycerol to 2.4 Å. The phosphate binds near the catalytic serine in a way that mimics the hemiacetal that develops during catalysis, while the glycerol occupies the side-chain binding pocket. Our structures show that the enzyme is not only structurally similar to penicillin G acylase but also employs essentially the same mechanism in which the ,-amino group of the catalytic serine acts as a base. A subtle difference is the presence of two catalytic dyads, His B23/Glu B455 and His B23/Ser B1, that are not seen in penicillin G acylase. In contrast to classical serine proteases, the central histidine of these dyads interacts indirectly with the O, through a hydrogen bond relay network involving the ,-amino group of the serine and a bound water molecule. A plausible model of the enzyme,substrate complex is proposed that leads to the prediction of mutants of glutarylamidase that should enable the enzyme to deacylate cephalosporin C into 7-aminocephalosporanic acid. [source] Downstream Processing of Enzymatically Produced Geranyl GlucosideBIOTECHNOLOGY PROGRESS, Issue 5 2001B. Mattheus de Roode Geraniol plays an important role in the fragrance and flavor industry. The corresponding glucoside has interesting properties as a "slow release" aroma compound. Therefore, the enzymatic production and downstream processing of geranyl glucoside were investigated. Geranyl glucoside was produced in a spray column reactor with an initial production rate of 0.58 mg U,1 h,1. A pretreated hydrophobic microfiltration membrane was used to prevent migration of the aqueous, enzyme-containing phase to the downstream process. No retention of the glucoside, which accumulated in the geraniol phase, was found. On the basis of examples from the literature, four downstream processes were tested on their viability for this system. Extraction with water and foaming were not suitable to recover geranyl glucoside from geraniol. In the first case, the glucoside selectivity for the geraniol phase was found to be high, which made extraction with water unsuccessful. In the second case it was possible to obtain a stable foam, but significant enrichment of the foam with glucoside did not occur. Adsorption on alumina and distillation under reduced pressure were applied successfully and tested in-line with the bioreactor. A maximum glucoside adsorption of 7.86 mg g,1 was achieved on alumina. After desorption and evaporation of the extractant the pure glucoside was obtained quantitatively. A pure product could not be obtained after distillation because a small amount of glucose was present in the permeate as well, which accumulated in the bottom fraction. It was shown that with this reactor system a production of 1 kg of geranyl glucoside in 2 days is possible using an initial amount of 50,000 units of enzyme. [source] | |||||||||||||