ENZYME TECHNOLOGY
INDUSTRIAL APPLICATIONS
fuels industry that involves enzymes depends largely on the ability of enzymes to develop efficient enough to be economically viable use.
current use of lignocellulose has become important for rapid progress in research for the production of bioethanol. By its content of hemicellulose, cellulose, lignin, extra-active and include organic and inorganic compounds, the lignocellulose raw material is suitable for the production of biofuel ethanol. In the pre-treatment processes used acid or fungi to facilitate the use of enzymes that degrade lignocellulose.
are then used enzymes from different sources mainly fungal hemicellulase and cellulase Trichoderma reesei , but further improvements were found by hydrolyzing esterases synergy with spergillus A niger and Pycnoporus cinnabarinus laccases of ; this objective undo hydrolysis is the compact and free sugars such as cellobiose, disaccharide . The use of the enzyme in the production of biofuels is generally rationed lipases, cellulases, hemicellulases and laccases. The table shows that the field premium for biofuels with these enzymes is waste vegetable oil and pray that contain cellulose, hemicellulose and lignin, such as bagasse or sugarcane residue, apart from certain plants that are grown for this purpose.
ENZYMES IN MOLECULAR BIOLOGY.
Restriction enzymes: EcoRI . -Restriction enzymes are a type of endonuclease that cut in a specific sequence, creating sticky and blunt ends. The bacteria used to protect against viruses, preventing their replication. Do not cut methylated regions.
Structure: homodimeric; Subunit: 4 chains consevadas 1 β, β 2, β 3, β 4 and a α-helix. Core: common center (catalytic)-spatial proximity of two carboxylates (aspartate yu aspartate or glutamate residue) and a lysine residue.
Applications: Genetic mapping, DNA sequencing (Sanger enzymatic put), DNA recombination.
DNA (Taq) polymerase. -A type of DNA polymerase that withstands high temperatures.
Structure: Klenow Fragment: Product hydrolytic of the enzyme E. coli DNA polymerase Natia exonuclease activity that lacks 5'-3 '. Homology with the native polymerase amino acid level (the site of addition of nucleotides 3'-OH, the binding sites dNTP / DNA and the sites 5'-3 'endonucleases. Transcriptase activity has no domain Evers and proofreading 3 '-5'.
Applications : Clinical diagnosis, amplification of RNA, DNA Finger-printing, PCR (RFLP)
DAIRY INDUSTRY.
1. enzymatic coagulation of casein .- The enzymatic coagulation of milk to obtain cheese is a process in which the casein, the main component of milk protein is denatured by the action of proteolytic enzymes. Casein is found in milk in the form of colloidal particles of calcium fosfocaseinato. These particles are in equilibrium with milk metoestable shaped micelles. It consists of 4 fractions: α -S1, -α S2, β and κ , and possibly others. Proteases cut the chain κ casein the link phe5-met-106 in the hydrophilic region from the end.
2. Developing Lactose-free products .- The use of β -galactosidase in the production of dairy products has increased very helpless. The main feature of these foods is that a lactose intolerant person can consume without any problems.
Advantages: Increased sweetness, solubility and fermentable sugars.
Making sugars resulting from hydrolysis of lactose allows better absorption of vitamins and minerals, especially calcium, avoiding with this scaling problem.
3. Technological Innovations .-
Immobilization of proteases;
L to microencapsulation of enzymes has proved a viable alternative to avoid early breakdown of milk caseins in cheese preparation.
Immobilization of β -galactosidase;
Methods: Silica gel coated with chitosan, agarose Tiosulfont, aluminum, fiberglass wool. Considerations: Decreases the effect of inhibition due to the low difference between substrate and product concentrations, low loss of enzyme due to the absence of collisions between enzyme and driver.
FOOD PROCESSING.
Advantages: Specificity (prevents side reactions), operate at ambient conditions, increase yields, improve product quality.
Limitations: Quality (toxicity or pathogenicity of the organism from which it draws), it is necessary to make a cost-benefit balance to assess whether it is better to use.
Current Innovations: Genetic engineering of microorganisms for processing to obtain new enzymes.
Applications:
Amylases .- Baked Goods, Beers, Increasing the fermentable sugars for yeast to produce maltose alcoholic fermentation.
Cellulase .- Cerveza; hydrolyze the cell wall of barley seeds and attachments.
.- Invertase sweeteners, sugars invested for the production of candies and syrups from sucrose.
Lactase .- Ice cream and milk; prevent crystallization of lactose with avoiding sandy texture also get milk suitable for lactose intolerant people.
Naraginasa .- Citrus juices, remove the bitter flavors, especially grapefruit.
pectic enzymes. -Chocolate, Cocoa, coffee, fruit juices, olive oil, hydrolytic activity during fermentation cocoa. Gelatinous covering hydrolyzed during the fermentation of the grapes. Increases the efficiency of extraction by pressing, clarification, improved concentration process.
Proteases .- bakery products, beer, meat and fish; Increase the extensibility of the dough, improve texture, crumb and loaf volume. Develop the body and flavor of beer to help clarification. Miso manufacture.
Lipases .- Lipids; convert fats and oils to glycerol and fatty acids and involved in trans-esterification reactions.
glucose oxidase and catalase .- Variety of products, to eliminate oxygen and / or glucose to prevent oxidation and browning.
and insurance regulation.
The application of enzymes in food is regulated, various international agencies like WHO and guidelines that have been studied and developed guidelines for the application of enzymes as additives or processes assistants. If enzymes are used either as additive or in the process, these agencies must ensure that the enzymes used obtained from non-pathogenic microorganisms and non-toxic and when the enzyme was obtained by recombinant DNA, the characteristics of the donor agency and proofreader are studied deeply. As for the labeling of foods if they are used as aides processes have no role in the final product, so no need to declare on the label, otherwise if necessary.
PHARMACEUTICAL INDUSTRY.
Importance: Production of new drugs quickly and efficiently, attacking understand the complex mechanisms of disease., to combat hereditary diseases and acquired Biochemical deficiencies related to the body, most drugs have ecological processes with low energy, high specificity and efficiency of products, reduced in production costs of drugs.
.- Improving Pharmaceutical Biocatalysis.
Biocatalysis: Processes with temperatures, pressures and moderate conditions similar to natural enzymes. High specificity of the enzymes in your product and high selectivity in terms enanitomeros ephemeral and regional isomerization selectivity. Faster in terms of reactions, the affinity of enzymes to their substrates and reduce the energy required for activation. And reduction steps in the process of drug manufacturing thanks to the technique of immobilization of enzymes and Russian. Thanks to recombinant DNA technology can generate the design of a catalyst.
Organic Synthesis: Processes with extreme temperatures and pressures that can cause problems with isomerization, enantiomers, epimers and re-arrangements. Low specificity and low efficiency by having products. Limited to the likely reactions and the molecules are and react. In most compounds processes are replaced by the high cost of having their separation after the reaction. Catalysis with heavy metals and radioactive compounds. applications. Management
enzymes for the production of drugs and their compuestos. "In this aspect the hydrolytic enzymes are widely used in organic synthesis as environmentally friendly catalysts with a broad substrate specificity, high stereo-selectivity, commercially available and do not require cofactors.
Use of enzymes as drugs to combat genetic diseases and deficiencies adquiridas. "These enzymes are produced to carry out the functions that the agency also stopped to make or break a molecule that is detrimental to the proper functioning of the body. Example: Anti-cancer, anti-inflammatory, anti-depressants, immuno-suppressants.
Innovations .- Directed evolution of enzymes by genetic engineering methods, DNA shuffling and HTP screening. Besides the use of gas phases and semi-aqueous solutions for enzyme catalysis. examples.
Neoplasms Control. -Intravenous administration of L-asparagine in the treatment of neoplasms that affect blood cells, as the case of acute lymphocytic leukemia. The mechanism for the use of this enzyme is based on the observation that certain tumor cells have a nutritional requirement for exogenous L-asparagine. Consequently, the administration of this enzyme decrease blood levels of L-asparagine causes a regression of the neoplasm.
Gaucher syndrome. -hereditary disease of the sphingolipid metabolism causes a build up of fatty deposits in certain organs and bones. In healthy individuals the α-glucocerebrosidase help break down fats and the glucocerebrosides. Gaucher cells accumulate in the liver, spleen and bone marrow. Ceredase ® (agulcerasa) is a modification of the β-glucocerebrosidase in the oligosaccharide chains of the enzyme. This modification alters the sugar residues of the glycoprotein chain to end with mannose residues that are recognized by carbohydrate receptors on macrophages. The main enzymatic source of human placental tissue.
bioremediation.
As the great nineteenth-century industrial development in terms of processes and applications, srios has led to environmental problems. Ultimames in decades emerged the concept of bioremediation through the use of biological agents. Bioremediation processes can be: phytoremediation, microbial remediation or enzymatic degradation.
Bioremediation enzymatic .- In the 60's comes this method to degrade zenobioticos and is now used in ndustrial processes such as textiles, food and detergents. The degradacón is carried out by enzymes obtained from mocroorganimos. At times, these are genetically modified by marketers to enhance the genes that express these enzymes.
Advantages: Wide range of environmental conditions, recoverable, acting with different substrates, biodegradable.
Disadvantages: High doses of enzymes if they are many, can not increase their number, some lose their reactivity, they have the ability to adapt.
Main biological functions. -degradable toxic fenores released during the degradation of lignin and cytokines produced by other agencies. Involved in lignin degradation with peroxidases.
Example .
Lactase EC.1.10.3.2 .- Obtained from Trametes versicolor, extracted from the extracellular region, works at a pH between 3 and 7 with optimum temperature of 30 ° C. Tiemo industrial applications such as the removal of dyes, bleaching paper and textile industry.
INDUSTRY DETERGENTS.
enzymes optimize the efficiency of detergents, while allowing the cleanup work at lower temperatures and shorter periods of washing, chose to reduce consumption and emissions of CO 2 . Another environmental benefit associated with the use of enzymes in detergents qes these are biodegradable and replace the chemicals of conventional detegentes. Associated enzymes in detergents are used mainly proteases, are also anqune amylases, lipases and cell Suad in lower proportion. These enzymes have been improved by protein engineering techniques or from microorganisms recombinant to optimize its manufacturing process.
The use of enzymes in detergents has the following functions: Minimize water use, reduce energy consumption and CO2 emissions, are biodegradable and reemmplazan to synthetic chemicals are released into the environment, allow cleanup work at lower temperatures and shorter periods of washing, occupy less than 1% of the total volume of detergent. Enzimpas thermophilic .- The first product of an extremophile used in the industry was the Cellulase 103, as an additive for detergents in 1997 as a bleaching agent.
Example .
Alkazyme .- A Thermocatenulatus Bacillus lipase expressed in E. coli, stable at pH 11 and a maximum activity at 45 ° C. The activity is over 90% within 5 minutes after the start of the wash cycle, the cellulase activity is below 50% within 5 to 10 minutes cycle Washing and less than 10% of residual cellulase activity is reached after 15 minutes in the cycle.
PAPER INDUSTRY.
The manufacture of pulp, paper and paper products is one of the largest industries in the world. Its main raw material is wood, chemically composed of cellulose, lignin (16-33%), hemicellulose and other materials (up to 10%). The Lignin is a complex aromatic polymer plant cell wall degradation is difficult, given its structure stereo-irregular , which represents industrial by-product that must be removed annoying timber during relief operations in the pulp. The only organisms that efficiently degrade this molecule are basidiomycetes fungi (eg Phanerochaete chrysosporium ) white rot.
The MONEY is to remove the residual lignin (which causes the brown discoloration) to avoid damaging the fiber quality, giving the white corresponding to the standards commercial. Although There are conventional methods such as (using Cl 2 , ClO 2 and NaOH), ECF (free Cl 2 ), TCF (total Cl-free) or treatment with ozone, peroxide and thermomechanical none represents many potential advantages as enzyme treatment. The objective of pulping is to remove lignin for pulp, either raw (Kraft) or bleached, contained in the wood. The process can be mechanical (crushing the fibers are wood, paper usually get weak and discolored) or Chemical (firing wood, high temperature and pressure, with white liquor composed of NaOH and Na 2 S), which are processes with high emissions of sulfur gases, so that more efficient methods are promoted as the Modified Continuous Cooking and pre-oxygenation delignification.
Bio-dispersants are compounds formulated for industrial purposes, used alone or with enzymes, which may prevent the formation of biofilms (lama of bacterial colonies). They work by slowing or preventing the formation and adhesion of micro-colonies of bacteria to surfaces. They can be mono-molecular compounds, polymers, or highly specialized formulations containing surfactants, organic dispersants, penetrants, chelating agents and stabilizers.
Current Research develop technologies from various enzymes that help the wood decay, such as xylanases (breaking chemical bonds between lignin wood) or laccases (cuproenzimas produced by fungi that directly remove lignin) which have the advantage of being produced in large scale bioreactors, unlike ligninolytic enzymes such as lignin / Mn-peroxidase.
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