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Enzymatic Steps (enzymatic + step)
Selected AbstractsFermentation of enzymatic hydrolysates from olive stones by Pachysolen tannophilusJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 3 2009Manuel Cuevas Abstract BACKGROUND: Olive stones were pretreated with liquid hot water (LHW or autohydrolysis) at maximum temperatures between 175 and 225 °C (severity factors, logR0, between 2.73 and 4.39) to be subjected (both liquid and solid components) afterwards to enzymatic hydrolysis with cellulases from Trichoderma viride. Ethanol fermentation of hydrolysates was performed with the non-traditional yeast Pachysolen tannophilus ATCC 32691. RESULTS: After the enzymatic step, yields of hemicellulose solubilization reached 100%, while the cellulose was only partially hydrolysed (23%, logR0 = 4.39). The maximum yields in total reducing sugars and acetic acid, at the upper end of the severity range, was close to 0.25 and 0.04 g g,1 dry stone, respectively. During the fermentation stage, the increase in R0 reduced the maximum specific growth rate, biomass productivity, and overall biomass yield. The overall yields of ethanol and xylitol ranged, respectively, from 0.18 to 0.25 g g,1 and from 0.01 to 0.13 g g,1. CONCLUSIONS: The results demonstrate the possibility of producing ethanol from olive stones, making use of the cellulose and hemicellulose fraction of the waste. It was confirmed that the overall yield in xylitol strongly depended on severity factor, while the overall yield in ethanol remained practically constant for all the pretreatment conditions tested. Copyright © 2008 Society of Chemical Industry [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] Roles of reactive oxygen species in the corpus luteumANIMAL SCIENCE JOURNAL, Issue 6 2006Norihiro SUGINO ABSTRACT Cells living under aerobic conditions always face the oxygen paradox. Oxygen is necessary for cells to maintain their lives. However, reactive oxygen species such as superoxide radicals, hydroxyl radicals and hydrogen peroxide are generated from oxygen and damage cells. Oxidative stress occurs as a consequence of the excessive production of reactive oxygen species and impaired antioxidant defense systems. Antioxidant enzymes include superoxide dismutase (SOD), which is a specific enzyme to scavenge superoxide radicals; copper-zinc SOD, located in the cytosol and Mn-SOD, located in the mitochondria. Both types of SOD belong to the first enzymatic step to scavenge superoxide radicals. It has been reported that a number of local factors such as cytokines, growth factors and eicosanoids are involved in the regulation of the corpus luteum (CL) function in addition to gonadotropins. Since reactive oxygen species are generated and SOD is expressed in the CL, there is a possibility that reactive oxygen species and SOD work as local regulators of the CL function. The present review reports that reactive oxygen species and their scavenging systems play important roles in the regulation of the CL function. [source] Combined Chemical-Enzymatic Assembly of Aminoglycoside Derivatives with N-1-AHB Side ChainADVANCED SYNTHESIS & CATALYSIS (PREVIOUSLY: JOURNAL FUER PRAKTISCHE CHEMIE), Issue 11-12 2008Igor Nudelman Abstract A series of unprotected pseudo-disaccharides and pseudo-trisaccharides of 2-deoxystreptamine-containing aminoglycosides have been selectively acylated at the N-1 position with the valuable (S)-4-amino-2-hydroxybutanoyl (AHB) pharmacophore by using the recombinant BtrH and BtrG enzymes from butirosin biosynthesis in combination with a synthetic acyl donor. The process was optimized by performing two enzymatic steps in a sequential manner without purification of the intermediate product. [source] Crystallization and preliminary X-ray analysis of adenylylsulfate reductase from Archaeoglobus fulgidusACTA CRYSTALLOGRAPHICA SECTION D, Issue 12 2000Annette Roth A group of anaerobic microorganisms use sulfate as the terminal electron acceptor for energy conservation. The process of sulfate reduction involves several enzymatic steps. One of them is the conversion of adenylyl sulfate (adenosine-5,-phosphosulfate) to sulfite, catalyzed by adenylylsulfate reductase. This enzyme is composed of a FAD-containing ,-subunit and a ,-subunit harbouring two [4Fe,4S] clusters. Adenylylsulfate reductase was isolated from Archaeoglobus fulgidus under anaerobic conditions and crystallized using the hanging-drop vapour-diffusion method using PEG 4000 as precipitant. The crystals grew in space group P212121, with unit-cell parameters a = 72.4, b = 113.2, c = 194.0,Å. The asymmetric unit probably contains two ,, units. The crystals diffract beyond 2,Å resolution and are suitable for X-ray structure analysis. [source] Real-time detection of the morphological change in cellulose by a nanomechanical sensorBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2010Liming Zhao Abstract Up to now, experimental limitations have prevented researchers from achieving the molecular-level understanding for the initial steps of the enzymatic hydrolysis of cellulose, where cellulase breaks down the crystal structure on the surface region of cellulose and exposes cellulose chains for the subsequent hydrolysis by cellulase. Because one of these non-hydrolytic enzymatic steps could be the rate-limiting step for the entire enzymatic hydrolysis of crystalline cellulose by cellulase, being able to analyze and understand these steps is instrumental in uncovering novel leads for improving the efficiency of cellulase. In this communication, we report an innovative application of the microcantilever technique for a real-time assessment of the morphological change of cellulose induced by a treatment of sodium chloride. This sensitive nanomechanical approach to define changes in surface structure of cellulose has the potential to permit a real-time assessment of the effect of the non-hydrolytic activities of cellulase on cellulose and thereby to provide a comprehensive understanding of the initial steps of the enzymatic hydrolysis of cellulose. Biotechnol. Bioeng. 2010;107: 190,194. © 2010 Wiley Periodicals, Inc. [source] Dopamine, Morphine, and Nitric Oxide: An Evolutionary Signaling TriadCNS: NEUROSCIENCE AND THERAPEUTICS, Issue 3 2010George B. Stefano Morphine biosynthesis in relatively simple and complex integrated animal systems has been demonstrated. Key enzymes in the biosynthetic pathway have also been identified, that is, CYP2D6 and COMT. Endogenous morphine appears to exert highly selective actions via novel mu opiate receptor subtypes, that is, mu3,-4, which are coupled to constitutive nitric oxide release, exerting general yet specific down regulatory actions in various animal tissues. The pivotal role of dopamine as a chemical intermediate in the morphine biosynthetic pathway in plants establishes a functional basis for its expansion into an essential role as the progenitor catecholamine signaling molecule underlying neural and neuroendocrine transmission across diverse animal phyla. In invertebrate neural systems, dopamine serves as the preeminent catecholamine signaling molecule, with the emergence and limited utilization of norepinephrine in newly defined adaptational chemical circuits required by a rapidly expanding set of physiological demands, that is, motor and motivational networks. In vertebrates epinephrine, emerges as the major end of the catecholamine synthetic pathway consistent with a newly incorporated regulatory modification. Given the striking similarities between the enzymatic steps in the morphine biosynthetic pathway and those driving the evolutionary adaptation of catecholamine chemical species to accommodate an expansion of interactive but distinct signaling systems, it is our overall contention that the evolutionary emergence of catecholamine systems required conservation and selective "retrofit" of specific enzyme activities, that is, COMT, drawn from cellular morphine expression. Our compelling hypothesis promises to initiate the reexamination of clinical studies, adding new information and treatment modalities in biomedicine. [source] | |||||||||||||