3-Isopropylmalate Dehydrogenase
Alcohol Oxidoreductases
Thermus thermophilus
2-Isopropylmalate Synthase
Thermus
Enzyme Stability
Malates
Isocitrate Dehydrogenase
Sulfolobus
Substrate Specificity
Molecular Sequence Data
Escherichia coli
Amino Acid Sequence
Mutagenesis, Site-Directed
Protein Denaturation
L-Lactate Dehydrogenase
Crystallography, X-Ray
Models, Molecular
Sequence Alignment
Protein Conformation
Cloning, Molecular
Alcohol Dehydrogenase
Base Sequence
Further improvement of the thermal stability of a partially stabilized Bacillus subtilis 3-isopropylmalate dehydrogenase variant by random and site-directed mutagenesis. (1/111)
A thermostabilized mutant of Bacillus subtilis 3-isopropylmalate dehydrogenase (IPMDH) obtained in a previous study contained a set of triple amino acid substitutions. To further improve the stability of the mutant, we used a random mutagenesis technique and identified two additional thermostabilizing substitutions, Thr22-->Lys and Met256-->Val, that separately endowed the protein with further stability. We introduced the two mutations into a single enzyme molecule, thus constructing a mutant with overall quintuple mutations. Other studies have suggested that an improved hydrophobic subunit interaction and a rigid type II beta-turn play important roles in enhancing the protein stability. Based on those observations, we successively introduced amino acid substitutions into the mutant with the quintuple mutations by site-directed mutagenesis: Glu253 at the subunit interface was replaced by Leu to increase the hydrophobic interaction between the subunits; Glu112, Ser113 and Ser115 that were involved in the formation of the turn were replaced by Pro, Gly and Glu, respectively, to make the turn more rigid. The thermal stability of the mutants was determined based on remaining activity after heat treatment and first-order rate constant of thermal unfolding, which showed gradual increases in thermal stability as more mutations were included. (+info)Functional analysis of upstream regulating regions from the Yarrowia lipolytica XPR2 promoter. (2/111)
The XPR2 gene from Yarrowia lipolytica encodes an inducible alkaline extracellular protease. Its complex regulation involves pH, carbon, nitrogen and peptones. Two previously identified upstream activating sequence (UAS) regions were analysed in a reporter system, outside the XPR2 context. Fragments from the UAS regions were inserted upstream of a minimal LEU2 promoter directing the expression of a reporter gene. The activity of the hybrid promoters was assessed following integration into the Y. lipolytica genome. This study confirmed the presence of two UASs composed of several interacting elements. Within the distal UAS (UAS1), a TUF/RAP1 binding site exhibited a UAS activity, which was enhanced by the presence of two adjacent repeats, overlapping sites similar to the CAR1 upstream repressing sequence from Saccharomyces cerevisiae. Within the proximal UAS (UAS2), the UAS activity required the interaction of both an ABF1-like binding site and a decameric repeat, containing Aspergillus nidulans PacC site consensus sequences. This decameric repeat was able to mediate repression due to carbon and/or nitrogen sources as well as pH-dependent activation. A study in the context of trans-regulatory mutations in the Y. lipolytica RIM101 gene showed that the PacC-like sites, potential binding sites for YlRim101p, were implicated in the derepression of UAS2-driven expression at neutral-alkaline pH. The in vivo response of the PacC-like decamers to external pH was dependent on the status of the pH-regulated activator YlRim101p, which is homologous to the A. nidulans PacC regulator. The carbon/nitrogen regulation imposed on the decamers was shown to be independent of YlRim101p and to override its effects. (+info)Escherichia coli Lrp (leucine-responsive regulatory protein) does not directly regulate expression of the leu operon promoter. (3/111)
Studies by R. Lin et al. (J. Bacteriol. 174:1948-1955, 1992) suggested that the Escherichia coli leu operon might be a member of the Lrp regulon. Their results were obtained with a leucine auxotroph; in leucine prototrophs grown in a medium lacking leucine, there was little difference in leu operon expression between lrp(+) and lrp strains. Furthermore, when leuP-lacZ transcriptional fusions that lacked the leu attenuator were used, expression from the leu promoter varied less than twofold between lrp(+) and lrp strains, irrespective of whether or not excess leucine was added to the medium. The simplest explanation of the observations of Lin et al. is that the known elevated leucine transport capacity of lrp strains (S. A. Haney et al., J. Bacteriol. 174:108-115, 1992) leads to very high intracellular levels of leucine for strains grown with leucine, resulting in the superattenuation of leu operon expression. (+info)Mirror image mutations reveal the significance of an intersubunit ion cluster in the stability of 3-isopropylmalate dehydrogenase. (4/111)
The comparison of the three-dimensional structures of thermophilic (Thermus thermophilus) and mesophilic (Escherichia coli) 3-isopropylmalate dehydrogenases (IPMDH, EC 1.1.1.85) suggested that the existence of extra ion pairs in the thermophilic enzyme found in the intersubunit region may be an important factor for thermostability. As a test of our assumption, glutamine 200 in the E. coli enzyme was turned into glutamate (Q200E mutant) to mimic the thermophilic enzyme at this site by creating an intersubunit ion pair which can join existing ion clusters. At the same site in the thermophilic enzyme we changed glutamate 190 into glutamine (E190Q), hereby removing the corresponding ion pair. These single amino acid replacements resulted in increased thermostability of the mesophilic and decreased thermostability of the thermophilic enzyme, as measured by spectropolarimetry and differential scanning microcalorimetry. (+info)Crystal structures of 3-isopropylmalate dehydrogenases with mutations at the C-terminus: crystallographic analyses of structure-stability relationships. (5/111)
Thermal stability of the Thermus thermophilus isopropylmalate dehydrogenase enzyme was substantially lost upon the deletion of three residues from the C-terminus. However, the stability was partly recovered by the addition of two, four and seven amino acid residues (called HD177, HD708 and HD711, respectively) to the C-terminal region of the truncated enzyme. Three structures of these mutant enzymes were determined by an X-ray diffraction method. All protein crystals belong to space group P2(1) and their structures were solved by a standard molecular replacement method where the original dimer structure of the A172L mutant was used as a search model. Thermal stability of these mutant enzymes is discussed based on the 3D structure with special attention to the width of the active-site groove and the minor groove, distortion of beta-sheet pillar structure and size of cavity in the domain-domain interface around the C-terminus. Our previous studies revealed that the thermal stability of isopropylmalate dehydrogenase increases when the active-site cleft is closed (the closed form). In the present study it is shown that the active-site cleft can be regulated by open-close movement of the minor groove located at the opposite side to the active-site groove on the same subunit, through a paperclip-like motion. (+info)Identification of enzymes homologous to isocitrate dehydrogenase that are involved in coenzyme B and leucine biosynthesis in methanoarchaea. (6/111)
Two putative Methanococcus jannaschii isocitrate dehydrogenase genes, MJ1596 and MJ0720, were cloned and overexpressed in Escherichia coli, and their gene products were tested for the ability to catalyze the NAD- and NADP-dependent oxidative decarboxylation of DL-threo-3-isopropylmalic acid, threo-isocitrate, erythro-isocitrate, and homologs of threo-isocitrate. Neither enzyme was found to use any of the isomers of isocitrate as a substrate. The protein product of the MJ1596 gene, designated AksF, catalyzed the NAD-dependent decarboxylation of intermediates in the biosynthesis of 7-mercaptoheptanoic acid, a moiety of methanoarchaeal coenzyme B (7-mercaptoheptanylthreonine phosphate). These intermediates included (-)-threo-isohomocitrate [(-)-threo-1-hydroxy-1,2, 4-butanetricarboxylic acid], (-)-threo-iso(homo)(2)citrate [(-)-threo-1-hydroxy-1,2,5-pentanetricarboxylic acid], and (-)-threo-iso(homo)(3)citrate [(-)-threo-1-hydroxy-1,2, 6-hexanetricarboxylic acid]. The protein product of MJ0720 was found to be alpha-isopropylmalate dehydrogenase (LeuB) and was found to catalyze the NAD-dependent decarboxylation of one isomer of DL-threo-isopropylmalate to 2-ketoisocaproate; thus, it is involved in the biosynthesis of leucine. The AksF enzyme proved to be thermostable, losing only 10% of its enzymatic activity after heating at 100 degrees C for 10 min, whereas the LeuB enzyme lost 50% of its enzymatic activity after heating at 80 degrees C for 10 min. (+info)The initial step of the thermal unfolding of 3-isopropylmalate dehydrogenase detected by the temperature-jump Laue method. (7/111)
A temperature-jump (T-jump) time-resolved X-ray crystallographic technique using the Laue method was developed to detect small, localized structural changes of proteins in crystals exposed to a temperature increase induced by laser irradiation. In a chimeric protein between thermophilic and mesophilic 3-isopropylmalate dehydrogenases (2T2M6T), the initial structural change upon T-jump to a denaturing temperature (approximately 90 degrees C) was found to be localized at a region which includes a beta-turn and a loop located between the two domains of the enzyme. A mutant, 2T2M6T-E110P/S111G/S113E, having amino acid replacements in this beta-turn region with the corresponding residues of the thermophilic enzyme, showed greater stability than the original chimera (increase of T:(m) by approximately 10 degrees C) and no T-jump-induced structural change in this region was detected by our method. These results indicate that thermal unfolding of the original chimeric enzyme, 2T2M6T, is triggered in this beta-turn region. (+info)Functional prediction: identification of protein orthologs and paralogs. (8/111)
Orthologs typically retain the same function in the course of evolution. Using beta-decarboxylating dehydrogenase family as a model, we demonstrate that orthologs can be confidently identified. The strategy is based on our recent findings that substitutions of only a few amino acid residues in these enzymes are sufficient to exchange substrate and coenzyme specificities. Hence, the few major specificity determinants can serve as reliable markers for determining orthologous or paralogous relationships. The power of this approach has been demonstrated by correcting similarity-based functional misassignment and discovering new genes and related pathways, and should be broadly applicable to other enzyme families. (+info)3-Isopropylmalate dehydrogenase (IPMDH) is an enzyme that plays a crucial role in the metabolic pathway known as leucine biosynthesis. This enzyme catalyzes the third step of this pathway, which involves the oxidative decarboxylation of 3-isopropylmalate to form 2-isopropylmalate, while simultaneously reducing NAD+ to NADH. The reaction is as follows:
3-Isopropylmalate + NAD+ -> 2-isopropylmalate + CO2 + NADH
The IPMDH enzyme is found in various organisms, including bacteria, yeast, and plants. In humans, defects or mutations in the gene encoding this enzyme can lead to a rare genetic disorder called 3-isopropylmalate dehydrogenase deficiency. This condition results in elevated levels of leucine and other intermediates in the leucine biosynthesis pathway, which can cause neurological symptoms such as developmental delay, seizures, and hypotonia (low muscle tone).
Alcohol oxidoreductases are a class of enzymes that catalyze the oxidation of alcohols to aldehydes or ketones, while reducing nicotinamide adenine dinucleotide (NAD+) to NADH. These enzymes play an important role in the metabolism of alcohols and other organic compounds in living organisms.
The most well-known example of an alcohol oxidoreductase is alcohol dehydrogenase (ADH), which is responsible for the oxidation of ethanol to acetaldehyde in the liver during the metabolism of alcoholic beverages. Other examples include aldehyde dehydrogenases (ALDH) and sorbitol dehydrogenase (SDH).
These enzymes are important targets for the development of drugs used to treat alcohol use disorder, as inhibiting their activity can help to reduce the rate of ethanol metabolism and the severity of its effects on the body.
'Thermus thermophilus' is not a medical term, but a scientific name for a species of bacteria. It is commonly used in molecular biology and genetics research. Here is the biological definition:
'Thermus thermophilus' is a gram-negative, rod-shaped, thermophilic bacterium found in hot springs and other high-temperature environments. Its optimum growth temperature ranges from 65 to 70°C (149-158°F), with some strains able to grow at temperatures as high as 85°C (185°F). The bacterium's DNA polymerase enzyme, Taq polymerase, is widely used in the Polymerase Chain Reaction (PCR) technique for amplifying and analyzing DNA. 'Thermus thermophilus' has a single circular chromosome and can also have one or more plasmids. Its genome has been fully sequenced, making it an important model organism for studying extremophiles and their adaptations to harsh environments.
2-Isopropylmalate synthase is an enzyme that catalyzes the condensation of a molecule of acetyl-CoA with a molecule of 3-isopropylmalate to form a molecule of 2-isopropylmalate. This reaction is part of the leucine biosynthesis pathway in bacteria, fungi, and plants. The enzyme is also known as 2-isopropylmalate isomerase-ligase or simply isopropylmalate synthase. It requires the cofactor CoA and is inhibited by leucine, a product of the pathway. Deficiency in this enzyme can lead to a rare genetic disorder called 2-isopropylmalate synthase deficiency, which is characterized by developmental delay, seizures, and metabolic acidosis.
"Thermus" is not a medical term, but rather a genus of bacteria that are capable of growing in extreme temperatures. These bacteria are named after the Greek word "therme," which means heat. They are commonly found in hot springs and deep-sea hydrothermal vents, where the temperature can reach up to 70°C (158°F).
Some species of Thermus have been found to produce enzymes that remain active at high temperatures, making them useful in various industrial applications such as molecular biology and DNA amplification techniques like polymerase chain reaction (PCR). However, Thermus itself is not a medical term or concept.
Enzyme stability refers to the ability of an enzyme to maintain its structure and function under various environmental conditions, such as temperature, pH, and the presence of denaturants or inhibitors. A stable enzyme retains its activity and conformation over time and across a range of conditions, making it more suitable for industrial and therapeutic applications.
Enzymes can be stabilized through various methods, including chemical modification, immobilization, and protein engineering. Understanding the factors that affect enzyme stability is crucial for optimizing their use in biotechnology, medicine, and research.
I'm sorry for any confusion, but "Malates" is not a recognized term in medical terminology. It's possible there may be a spelling mistake or it could be a slang term or an abbreviation that is not widely recognized. If you have more context or information, I'd be happy to try and help further.
Isocitrate Dehydrogenase (IDH) is an enzyme that catalyzes the oxidative decarboxylation of isocitrate to α-ketoglutarate in the presence of NAD+ or NADP+, producing NADH or NADPH respectively. This reaction occurs in the citric acid cycle, also known as the Krebs cycle or tricarboxylic acid (TCA) cycle, which is a crucial metabolic pathway in the cell's energy production and biosynthesis of various molecules. There are three isoforms of IDH found in humans: IDH1 located in the cytosol, IDH2 in the mitochondrial matrix, and IDH3 within the mitochondria. Mutations in IDH1 and IDH2 have been associated with several types of cancer, such as gliomas and acute myeloid leukemia (AML), leading to abnormal accumulation of 2-hydroxyglutarate, which can contribute to tumorigenesis.
In a medical context, "hot temperature" is not a standard medical term with a specific definition. However, it is often used in relation to fever, which is a common symptom of illness. A fever is typically defined as a body temperature that is higher than normal, usually above 38°C (100.4°F) for adults and above 37.5-38°C (99.5-101.3°F) for children, depending on the source.
Therefore, when a medical professional talks about "hot temperature," they may be referring to a body temperature that is higher than normal due to fever or other causes. It's important to note that a high environmental temperature can also contribute to an elevated body temperature, so it's essential to consider both the body temperature and the environmental temperature when assessing a patient's condition.
"Sulfolobus" is a genus of archaea, which are single-celled microorganisms that share characteristics with both bacteria and eukaryotes. These archaea are extremophiles, meaning they thrive in extreme environments that are hostile to most other life forms. Specifically, Sulfolobus species are acidothermophiles, capable of growing at temperatures between 75-85°C and pH levels near 3. They are commonly found in volcanic hot springs and other acidic, high-temperature environments. The cells of Sulfolobus are typically irregular in shape and have a unique system for replicating their DNA. Some species are capable of oxidizing sulfur compounds as a source of energy.
Substrate specificity in the context of medical biochemistry and enzymology refers to the ability of an enzyme to selectively bind and catalyze a chemical reaction with a particular substrate (or a group of similar substrates) while discriminating against other molecules that are not substrates. This specificity arises from the three-dimensional structure of the enzyme, which has evolved to match the shape, charge distribution, and functional groups of its physiological substrate(s).
Substrate specificity is a fundamental property of enzymes that enables them to carry out highly selective chemical transformations in the complex cellular environment. The active site of an enzyme, where the catalysis takes place, has a unique conformation that complements the shape and charge distribution of its substrate(s). This ensures efficient recognition, binding, and conversion of the substrate into the desired product while minimizing unwanted side reactions with other molecules.
Substrate specificity can be categorized as:
1. Absolute specificity: An enzyme that can only act on a single substrate or a very narrow group of structurally related substrates, showing no activity towards any other molecule.
2. Group specificity: An enzyme that prefers to act on a particular functional group or class of compounds but can still accommodate minor structural variations within the substrate.
3. Broad or promiscuous specificity: An enzyme that can act on a wide range of structurally diverse substrates, albeit with varying catalytic efficiencies.
Understanding substrate specificity is crucial for elucidating enzymatic mechanisms, designing drugs that target specific enzymes or pathways, and developing biotechnological applications that rely on the controlled manipulation of enzyme activities.
Molecular sequence data refers to the specific arrangement of molecules, most commonly nucleotides in DNA or RNA, or amino acids in proteins, that make up a biological macromolecule. This data is generated through laboratory techniques such as sequencing, and provides information about the exact order of the constituent molecules. This data is crucial in various fields of biology, including genetics, evolution, and molecular biology, allowing for comparisons between different organisms, identification of genetic variations, and studies of gene function and regulation.
'Escherichia coli' (E. coli) is a type of gram-negative, facultatively anaerobic, rod-shaped bacterium that commonly inhabits the intestinal tract of humans and warm-blooded animals. It is a member of the family Enterobacteriaceae and one of the most well-studied prokaryotic model organisms in molecular biology.
While most E. coli strains are harmless and even beneficial to their hosts, some serotypes can cause various forms of gastrointestinal and extraintestinal illnesses in humans and animals. These pathogenic strains possess virulence factors that enable them to colonize and damage host tissues, leading to diseases such as diarrhea, urinary tract infections, pneumonia, and sepsis.
E. coli is a versatile organism with remarkable genetic diversity, which allows it to adapt to various environmental niches. It can be found in water, soil, food, and various man-made environments, making it an essential indicator of fecal contamination and a common cause of foodborne illnesses. The study of E. coli has contributed significantly to our understanding of fundamental biological processes, including DNA replication, gene regulation, and protein synthesis.
An amino acid sequence is the specific order of amino acids in a protein or peptide molecule, formed by the linking of the amino group (-NH2) of one amino acid to the carboxyl group (-COOH) of another amino acid through a peptide bond. The sequence is determined by the genetic code and is unique to each type of protein or peptide. It plays a crucial role in determining the three-dimensional structure and function of proteins.
Site-directed mutagenesis is a molecular biology technique used to introduce specific and targeted changes to a specific DNA sequence. This process involves creating a new variant of a gene or a specific region of interest within a DNA molecule by introducing a planned, deliberate change, or mutation, at a predetermined site within the DNA sequence.
The methodology typically involves the use of molecular tools such as PCR (polymerase chain reaction), restriction enzymes, and/or ligases to introduce the desired mutation(s) into a plasmid or other vector containing the target DNA sequence. The resulting modified DNA molecule can then be used to transform host cells, allowing for the production of large quantities of the mutated gene or protein for further study.
Site-directed mutagenesis is a valuable tool in basic research, drug discovery, and biotechnology applications where specific changes to a DNA sequence are required to understand gene function, investigate protein structure/function relationships, or engineer novel biological properties into existing genes or proteins.
Protein denaturation is a process in which the native structure of a protein is altered, leading to loss of its biological activity. This can be caused by various factors such as changes in temperature, pH, or exposure to chemicals or radiation. The three-dimensional shape of a protein is crucial for its function, and denaturation causes the protein to lose this shape, resulting in impaired or complete loss of function. Denaturation is often irreversible and can lead to the aggregation of proteins, which can have negative effects on cellular function and can contribute to diseases such as Alzheimer's and Parkinson's.
L-Lactate Dehydrogenase (LDH) is an enzyme found in various tissues within the body, including the heart, liver, kidneys, muscles, and brain. It plays a crucial role in the process of energy production, particularly during anaerobic conditions when oxygen levels are low.
In the presence of the coenzyme NADH, LDH catalyzes the conversion of pyruvate to lactate, generating NAD+ as a byproduct. Conversely, in the presence of NAD+, LDH can convert lactate back to pyruvate using NADH. This reversible reaction is essential for maintaining the balance between lactate and pyruvate levels within cells.
Elevated blood levels of LDH may indicate tissue damage or injury, as this enzyme can be released into the circulation following cellular breakdown. As a result, LDH is often used as a nonspecific biomarker for various medical conditions, such as myocardial infarction (heart attack), liver disease, muscle damage, and certain types of cancer. However, it's important to note that an isolated increase in LDH does not necessarily pinpoint the exact location or cause of tissue damage, and further diagnostic tests are usually required for confirmation.
X-ray crystallography is a technique used in structural biology to determine the three-dimensional arrangement of atoms in a crystal lattice. In this method, a beam of X-rays is directed at a crystal and diffracts, or spreads out, into a pattern of spots called reflections. The intensity and angle of each reflection are measured and used to create an electron density map, which reveals the position and type of atoms in the crystal. This information can be used to determine the molecular structure of a compound, including its shape, size, and chemical bonds. X-ray crystallography is a powerful tool for understanding the structure and function of biological macromolecules such as proteins and nucleic acids.
Molecular models are three-dimensional representations of molecular structures that are used in the field of molecular biology and chemistry to visualize and understand the spatial arrangement of atoms and bonds within a molecule. These models can be physical or computer-generated and allow researchers to study the shape, size, and behavior of molecules, which is crucial for understanding their function and interactions with other molecules.
Physical molecular models are often made up of balls (representing atoms) connected by rods or sticks (representing bonds). These models can be constructed manually using materials such as plastic or wooden balls and rods, or they can be created using 3D printing technology.
Computer-generated molecular models, on the other hand, are created using specialized software that allows researchers to visualize and manipulate molecular structures in three dimensions. These models can be used to simulate molecular interactions, predict molecular behavior, and design new drugs or chemicals with specific properties. Overall, molecular models play a critical role in advancing our understanding of molecular structures and their functions.
In genetics, sequence alignment is the process of arranging two or more DNA, RNA, or protein sequences to identify regions of similarity or homology between them. This is often done using computational methods to compare the nucleotide or amino acid sequences and identify matching patterns, which can provide insight into evolutionary relationships, functional domains, or potential genetic disorders. The alignment process typically involves adjusting gaps and mismatches in the sequences to maximize the similarity between them, resulting in an aligned sequence that can be visually represented and analyzed.
Protein conformation refers to the specific three-dimensional shape that a protein molecule assumes due to the spatial arrangement of its constituent amino acid residues and their associated chemical groups. This complex structure is determined by several factors, including covalent bonds (disulfide bridges), hydrogen bonds, van der Waals forces, and ionic bonds, which help stabilize the protein's unique conformation.
Protein conformations can be broadly classified into two categories: primary, secondary, tertiary, and quaternary structures. The primary structure represents the linear sequence of amino acids in a polypeptide chain. The secondary structure arises from local interactions between adjacent amino acid residues, leading to the formation of recurring motifs such as α-helices and β-sheets. Tertiary structure refers to the overall three-dimensional folding pattern of a single polypeptide chain, while quaternary structure describes the spatial arrangement of multiple folded polypeptide chains (subunits) that interact to form a functional protein complex.
Understanding protein conformation is crucial for elucidating protein function, as the specific three-dimensional shape of a protein directly influences its ability to interact with other molecules, such as ligands, nucleic acids, or other proteins. Any alterations in protein conformation due to genetic mutations, environmental factors, or chemical modifications can lead to loss of function, misfolding, aggregation, and disease states like neurodegenerative disorders and cancer.
Molecular cloning is a laboratory technique used to create multiple copies of a specific DNA sequence. This process involves several steps:
1. Isolation: The first step in molecular cloning is to isolate the DNA sequence of interest from the rest of the genomic DNA. This can be done using various methods such as PCR (polymerase chain reaction), restriction enzymes, or hybridization.
2. Vector construction: Once the DNA sequence of interest has been isolated, it must be inserted into a vector, which is a small circular DNA molecule that can replicate independently in a host cell. Common vectors used in molecular cloning include plasmids and phages.
3. Transformation: The constructed vector is then introduced into a host cell, usually a bacterial or yeast cell, through a process called transformation. This can be done using various methods such as electroporation or chemical transformation.
4. Selection: After transformation, the host cells are grown in selective media that allow only those cells containing the vector to grow. This ensures that the DNA sequence of interest has been successfully cloned into the vector.
5. Amplification: Once the host cells have been selected, they can be grown in large quantities to amplify the number of copies of the cloned DNA sequence.
Molecular cloning is a powerful tool in molecular biology and has numerous applications, including the production of recombinant proteins, gene therapy, functional analysis of genes, and genetic engineering.
Alcohol dehydrogenase (ADH) is a group of enzymes responsible for catalyzing the oxidation of alcohols to aldehydes or ketones, and reducing equivalents such as NAD+ to NADH. In humans, ADH plays a crucial role in the metabolism of ethanol, converting it into acetaldehyde, which is then further metabolized by aldehyde dehydrogenase (ALDH) into acetate. This process helps to detoxify and eliminate ethanol from the body. Additionally, ADH enzymes are also involved in the metabolism of other alcohols, such as methanol and ethylene glycol, which can be toxic if allowed to accumulate in the body.
'Bacillus subtilis' is a gram-positive, rod-shaped bacterium that is commonly found in soil and vegetation. It is a facultative anaerobe, meaning it can grow with or without oxygen. This bacterium is known for its ability to form durable endospores during unfavorable conditions, which allows it to survive in harsh environments for long periods of time.
'Bacillus subtilis' has been widely studied as a model organism in microbiology and molecular biology due to its genetic tractability and rapid growth. It is also used in various industrial applications, such as the production of enzymes, antibiotics, and other bioproducts.
Although 'Bacillus subtilis' is generally considered non-pathogenic, there have been rare cases of infection in immunocompromised individuals. It is important to note that this bacterium should not be confused with other pathogenic species within the genus Bacillus, such as B. anthracis (causative agent of anthrax) or B. cereus (a foodborne pathogen).
A base sequence in the context of molecular biology refers to the specific order of nucleotides in a DNA or RNA molecule. In DNA, these nucleotides are adenine (A), guanine (G), cytosine (C), and thymine (T). In RNA, uracil (U) takes the place of thymine. The base sequence contains genetic information that is transcribed into RNA and ultimately translated into proteins. It is the exact order of these bases that determines the genetic code and thus the function of the DNA or RNA molecule.
In the context of medicine and pharmacology, "kinetics" refers to the study of how a drug moves throughout the body, including its absorption, distribution, metabolism, and excretion (often abbreviated as ADME). This field is called "pharmacokinetics."
1. Absorption: This is the process of a drug moving from its site of administration into the bloodstream. Factors such as the route of administration (e.g., oral, intravenous, etc.), formulation, and individual physiological differences can affect absorption.
2. Distribution: Once a drug is in the bloodstream, it gets distributed throughout the body to various tissues and organs. This process is influenced by factors like blood flow, protein binding, and lipid solubility of the drug.
3. Metabolism: Drugs are often chemically modified in the body, typically in the liver, through processes known as metabolism. These changes can lead to the formation of active or inactive metabolites, which may then be further distributed, excreted, or undergo additional metabolic transformations.
4. Excretion: This is the process by which drugs and their metabolites are eliminated from the body, primarily through the kidneys (urine) and the liver (bile).
Understanding the kinetics of a drug is crucial for determining its optimal dosing regimen, potential interactions with other medications or foods, and any necessary adjustments for special populations like pediatric or geriatric patients, or those with impaired renal or hepatic function.
3-Isopropylmalate dehydrogenase
Isopropylmalic acid
Isocitrate dehydrogenase
List of MeSH codes (D08)
Gottfriedia
Branched-chain amino acid
Amino acid synthesis
List of EC numbers (EC 1)
Leucine
List of EC numbers (EC 2)
3-Isopropylmalate dehydrogenase - Wikipedia
3-isopropylmalate dehydrogenase (P04173) - Yeast Metabolome Database
Further stabilization of 3-isopropylmalate dehydrogenase of an extreme thermophile, Thermus thermophilus, by a suppressor...
SCOPe 2.07: Domain d1v53a : 1v53 A
Sequence search - BRENDA Enzyme Database
Untitled
Chorismate mutase of Thermus thermophilus is a monofunctional AroH class enzyme inhibited by tyrosine | SpringerLink
MeSH Browser
Finding step leuB for L-leucine biosynthesis in Shewanella loihica PV-4
Leucine. Medical search. Definitions
Browse
SA1863 - AureoWiki
1XCO | Genus
Regulon of T-box(Leu) in Bacillus amyloliquefaciens FZB42
Molecular genetic analysis of the region containing the essential Pseudomonas aeruginosa asd gene encoding aspartate-β...
SustainPineDB
Agaricus bisporus var bisporus H97
txid1368695[Organism:noexp] - Nucleotide - NCBI
Curated BLAST
Global transcriptional control by glucose and
BiGG Metabolite 3c2hmp c in iSbBS512 1146
MESH TREE NUMBER CHANGES - 2014 MeSH. July 29, 2013
MESH TREE NUMBER CHANGES - 2014 MeSH. July 29, 2013
MESH TREE NUMBER CHANGES - 2014 MeSH. July 29, 2013
MESH TREE NUMBER CHANGES - 2014 MeSH. July 29, 2013
MESH TREE NUMBER CHANGES - 2014 MeSH. July 29, 2013
MESH TREE NUMBER CHANGES - 2014 MeSH. July 29, 2013
MESH TREE NUMBER CHANGES - 2014 MeSH. July 29, 2013
1.851
- 3-Isopropylmalate dehydrogenase (EC 1.1.1.85) is an enzyme that is a part of the isopropylmalate dehydrogenase family, which catalyzes the chemical reactions: (2R,3S)-3-isopropylmalate + NAD+ ⇌ {\displaystyle \rightleftharpoons } 4-methyl-2-oxopentanoate + CO2 + NADH (2R,3S)-3-isopropylmalate + NAD+ ⇌ {\displaystyle \rightleftharpoons } (2S)-2-isopropyl-3-oxosuccinate + H+ + NADH (2S)-2-isopropyl-3-oxosuccinate + H+ ⇌ {\displaystyle \rightleftharpoons } 4-methyl-2-oxopentanoate + CO2 Burns RO, Umbarger HE, Gross SR (1963). (wikipedia.org)
Stability of 3-isopropylmalate dehydrogenase1
- We succeeded in further improvement of the stability of 3-isopropylmalate dehydrogenase (IPMDH) from an extreme thermophile, Thermus thermophilus, by a suppressor mutation method. (elsevierpure.com)
Aldehyde1
- Aldehyde dehydrogenase family, Iron-containing alcohol dehydrogenase [Interproscan]. (ntu.edu.sg)
Gene1
- Nucleotide sequence of the 3' terminal region of the LEU2 gene from Saccharomyces cerevisiae. (ymdb.ca)
Interproscan3
- short chain dehydrogenase [Interproscan]. (ntu.edu.sg)
- Glyceraldehyde 3-phosphate dehydrogenase [Interproscan]. (ntu.edu.sg)
- Glucose-6-phosphate dehydrogenase, NAD-binding [Interproscan]. (ntu.edu.sg)
LEUCINE1
- Leucine dehydrogenase catalyzes the reversible oxidative deamination of L-LEUCINE, to 4-methyl-2-oxopentanoate (2-ketoisocaproate) and AMMONIA, with the corresponding reduction of the cofactor NAD+. (lookformedical.com)
Pyruvate1
- protein_coding" "AAC74323","adhE","Escherichia coli","fused acetaldehyde-CoA dehydrogenase/iron-dependent alcohol dehydrogenase/pyruvate-formate lyase deactivase [Ensembl]. (ntu.edu.sg)
Superfamily1
- An octameric enzyme belonging to the superfamily of amino acid dehydrogenases. (lookformedical.com)
IDH11
- IDH1, also named as PICD and IDP, belongs to the isocitrate and isopropylmalate dehydrogenases family. (ptglab.com)
Sequences1
- After 3 days, severe diarrhea, stomach best matches for the sequences derived from Campy- ache, and shivering developed in the only 3 persons (the lobacter jejuni , but no sequences of the recovery DNA patient plus 2 family members) who had eaten undercooked sample showed any such signifi cant matches. (cdc.gov)
Cytochrome1
- They are further classified according to the acceptor which can be NAD+ or NADP+ (subclass 1.1.1), cytochrome (1.1.2), oxygen (1.1.3), quinone (1.1.5), or another acceptor (1.1.99). (curehunter.com)
Enzymes1
- A subclass of enzymes which includes all dehydrogenases acting on primary and secondary alcohols as well as hemiacetals. (curehunter.com)
Catalyzes1
- Catalyzes the oxidation of 3-carboxy-2-hydroxy-4- methylpentanoate (3-isopropylmalate) to 3-carboxy-4-methyl-2- oxopentanoate. (ymdb.ca)
19962
- 1996 Feb;178(3):723-727. (elsevierpure.com)
- 3, 137-155 (1996) REFERENCE 9 AUTHORS Fujita,N., Mori,H., Yura,T. and Ishihama,A. TITLE Systematic sequencing of the Escherichia coli genome: analysis of the 2.4-4.1 min (110,917-193,643 bp) region JOURNAL Nucleic Acids Res. (nig.ac.jp)
Major1
- This graph shows the total number of publications written about "Alcohol Dehydrogenase" by people in this website by year, and whether "Alcohol Dehydrogenase" was a major or minor topic of these publications. (wakehealth.edu)
Synthase1
- Initially, the L-leucine synthesis pathway was enhanced by overexpressing feedback-resistant 2-isopropylmalate synthase and acetohydroxy acid synthase both derived from Corynebacterium glutamicum, along with two other native enzymes. (bvsalud.org)
Oxidoreductase2
- Fungal members of this superfamily that are characterised and relevant for lignocellulose degradation include aryl-alcohol oxidoreductase, alcohol oxidase, cellobiose dehydrogenase, glucose oxidase, glucose dehydrogenase, pyranose dehydrogenase, and pyranose oxidase, which together form family AA3 of the auxiliary activities in the CAZy database of carbohydrate-active enzymes. (biomedcentral.com)
- Finally, we interpret our results from an evolutionary perspective, where we could show, for example, that pyranose dehydrogenase evolved from aryl-alcohol oxidoreductase after a change in substrate specificity and that the cytochrome domain of cellobiose dehydrogenase was regularly lost during evolution. (biomedcentral.com)
NADH1
- Then, the redox flux was improved by substituting the native NADPH-dependent acetohydroxy acid isomeroreductase, branched chain amino acid transaminase, and glutamate dehydrogenase with their NADH-dependent equivalents. (bvsalud.org)
Isocitrate3
- Isocitrate Dehydrogenases (IDHs) are important enzymes present in all living cells. (biomedcentral.com)
- Isocitrate Dehydrogenase (IDH) enzymes convert isocitrate to oxoglutarate in most living organisms. (biomedcentral.com)
- Isocitrate Dehydrogenases are important enzymes essential for survival of all organisms. (biomedcentral.com)
Escherichia2
- In an effort to characterize the properties that would improve its activity at temperatures less than the optimal, we subjected the thermostable Sulfolobus tokodaii (S. tokodaii) 3-isopropylmalate dehydrogenase to two rounds of random mutagenesis and selected for improved low-temperature activity using an in vivo recombinant Escherichia coli system. (elsevierpure.com)
- The glucose-methanol-choline (GMC) superfamily of oxidoreductases was defined by Cavener in 1992 based on sequence similarities of Drosophila melanogaster glucose dehydrogenase, Escherichia coli choline dehydrogenase, Aspergillus niger glucose oxidase, and Hansenula polymorpha methanol (alcohol) oxidase [ 12 ]. (biomedcentral.com)
Leucine Biosynthesis1
- As an intermediate for valine and leucine biosynthesis, 2-ketoisovalerate is decarboxylated by a heterologously expressed broad-substrate-range α-ketoisovalerate decarboxylase (Kivd) to isobutyaldehyde, and subsequently reduced into IB by an alcohol dehydrogenase (Adh). (biomedcentral.com)
Gene2
- In contrast, the homology between the 3-IMDHs of C. utilis and Thermus thermophilus was much smaller and was restricted to some regions of the gene. (microbiologyresearch.org)
- Nucleotide sequence of the 3′-terminal region of the LEU2 gene from Saccharomyces cerevisiae. (microbiologyresearch.org)
Protein2
- or (3) protein requirements are substantially increased as with extensive burns. (ncats.io)
- protein_coding" "Cz05g23010.t1","No alias","Chromochloris zofingiensis","4-hydroxy-3-methylbut-2-enyl diphosphate reductase [Interproscan]. (ntu.edu.sg)
Amino acids2
- An open reading frame of 1089 bp that corresponded to a polypeptide of 363 amino acids, one residue shorter than the 3-IMDH of S. cerevisiae , was found in the cloned fragment. (microbiologyresearch.org)
- The homology between the 3-IMDHs of C. utilis and S. cerevisiae was 76·2 % in nucleotides and 85·4 % in amino acids. (microbiologyresearch.org)
Family2
- After 3 days, severe diarrhea, stomach best matches for the sequences derived from Campy- ache, and shivering developed in the only 3 persons (the lobacter jejuni , but no sequences of the recovery DNA patient plus 2 family members) who had eaten undercooked sample showed any such signifi cant matches. (cdc.gov)
- part of family of 3'-to5' exonucleases. (lbl.gov)
Carbon1
- Isobutanol and 3-methyl-1-butanol production was effectively improved when overexpressing selected genes of the central carbon metabolism. (biomedcentral.com)
Term1
- Long-term cultivation was performed on two selected strains resulting in maximum cumulative isobutanol and 3-methyl-1-butanol titers of 1247 mg L −1 and 389 mg L −1 , on day 58 and day 48, respectively. (biomedcentral.com)
19971
- 1997 Mar;143(3):899-907. (nau.edu)
Properties1
- Five dehydrogenase mutants were purified and their catalytic properties and thermostabilities characterized. (elsevierpure.com)
Domain1
- Semialdehyde dehydrogenase, dimerisation domain [Interproscan]. (ntu.edu.sg)
Volume1
- Overexpression of NAF-1 enhanced antioxidant capacity and oxidative stress resistance, increasing tumor volume 3-fold. (alliedacademies.org)