1. What is enzyme immobilization?
Ans: Enzyme immobilization is defined as, restriction or prevention of movement or mobility of enzyme in a fixed space
2. What are the advantage of enzyme immobilization?
Ans: Reusability The immobilized enzymes can be easily recovered from the reaction mixture and can be reused because the enzyme is not consumed in the reaction and enzyme are also expensive and cannot be allowed to discard.
Pure Product The immobilized enzymes are used in any reaction at that time the product is recovered in pure state which is reduce the expensive of purification process.
3. Explain immobilization of enzymes by adsorption technique?
Ans: Adsorption involves the physical binding of enzymes or cells on the surface of inert support. The support materials may be inorganic (Aluminium,silica gel, calcium phosphate gel), Organic materials (Carboxy methyl cellulose). Adsorption of enzyme molecules involves weak forces and hydrogen bonds. Therefore the adsorbed enzymes can be easily removed by minor changes in Ph, ionic strength or temperature. This is an advantage of or industrial use of enzymes.
4. What is covalent binding?
Ans: Immobilization of enzymes can be achieved by creation of covalent bonds between chemical groups of enzymes and chemical groups of support material.
5. What is Diazotation?
Ans Some of the support materials such as aminobenzyle cellulose, amino derivatives of polystyrene, aminosilanized porus glass these are undergoes diazotation on treatment with sodium nitrate and hydrochloric acid, they turn and covalently bind to tyrosyl or histidyl groups of enzymes.
6. What is Biosensor?
Ans: Biosensor is an analytical device, used for the detection of a chemical substance, which combines a biological component with a physicochemical detector.
7. Write the advantages of Biosensors?
Ans: The key benefits of biosensors include the following
- Rapid and continuous measurement
- High specificity
- Very less usage of reagents required for calibration
- Fast response time
8.Write the principle involved in the working of biosensors?
Ans: Biosensors are operated based on the principle of signal transduction.
9. What is Protein engineering?
Ans: Protein Engineering is the process of developing proteins with desired function by manipulating stability and specificity of a protein. Protein engineering is the process of developing useful or valuable proteins. protein engineering is a method of changing a protein sequence to achieve a desired result, such as a change in the substrate specificity or increase stability to the temperature, organic solvents, and/ or extremes of pH
10.What are the main approaches of Protein Engineering?
Ans: There are two main approaches for protein engineering.
- Rational design or site-directed mutagenesis.
- Directed evolution or random mutagenesis
11.What is DNA shuffling?
Ans DNA shuffling is also known as sexual PCR. Before beginning with DNA shuffling, a pool of closely related molecules, with different point mutation techniques such as oligonucleotide-Directed mutagenesis.
12. What is Bioleachin?
Ans: Microbes are also used in a process called bioleaching, in which bacteria leach metals such as iron and manganese from soil and sewage. Bioleaching can change sediment structure, as well as create the potential to control water flow in aquifers and produce biomaterials.
13. Write the formulation of medium for production of enzymes?
Ans: The culture medium chosen should contain all the nutrients to support adequate growth of microorganism that will ultimately result in good quantities of enzymes production. The ingredients of the medium should be readily available at low cost and are nutritionally safe. Some of the commonly used substrates for the medium are starch hydrolysate, molasses, corn steep liquor, yeast extract, whey, and soy bean meal. Some cereals and pulses have also been used. The pH of the medium should be kept optimal should be kept optimal for good microbial growth and enzyme production.
14. How to produce precipitation of enzymes ?
Ans: Enzymes can be precipitated by using salts, organic solvents, precipitation is advantageous since the precipitated enzyme can be dissolved in a minimal volume to concentrate thee enzyme.
15. What is Amylase?
Ans: Amylase are a complex group of enzymes that hydrolyse polysaccharides like starch and glycogen to glucose. During the hydrolysis of 1, 4-glycoside linkage present in above polysaccharides are degraded which results first in the formation of shorts chain dextrins, then to maltose and glucose
16. What is DNA ligase? whose was it first discovered?
Ans: DNA ligase is a specific type of enzyme, a ligase, which facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. The first DNA ligase was purified and characterized in 196 by the Gellert, Lehman, Richardson, and Hurwitz laboratories.
17. What is Gel electrophoresis?
Ans: Gel electrophoresis is a routinely used analytical technique for the separation or purification of specific DNA fragments.
18. What is chimeric DNA?
Ans: Recombinent DNA molecules are sometimes called as chimeric DNA.
19. What are linkers?
Ans: Linkers are chemically synthesized, short double stranded DNA molecules.
20.Mention the basic technique in genetic engineering?
Ans: Agarose gel electrophoresis
- Isolation and purification and nucleic acids.
- Isolation of chromosomes
- Nucleic acid blotting techniques
- DNA sequencing
- Methods of gene transfer.
Ans: Electrophoresis is defined as movement of charged molecules in an electric field. The negatively charged molecules moves towards the positive electrode and positively charged molecules migrates towards negative electrode.
22. What are Plasmids?
Ans: Plasmids are extra chromosomal, double stranded, circular self replicating DNA molecules.
23.Write a note on RES?
Ans: Restriction endonucleases are one of the most important groups of enzymes. These are mainly useful for manipulation of DNA. These are the bacterial enzymes, used in cutting or splitting of DNA at specific sites. The first Restriction endonucleases was discovered by Hamilton smith in 190 in Hemophilus influenzae bacteria.
24. Write a note on alkaline phosphatase?
Ans: This enzyme is involved in removal of phosphate groups. This is useful to prevent the unwanted ligation of DNA molecules, this is the main problem occurred in cloning experiments. When the linear vector plasmid DNA is treated with alkaline phosphatase enzyme at that time the 5 terminal phosphate is removed. This prevents the recirculation and plasmid DNA dimer formulation at this time possible to inseration of foreign DNA through DNA ligase.
25. What are linkers and adapters?
Ans: These are chemically synthesized, short double stranded DNA molecules. These linkers possesses restriction enzyme on cleavage site, they can ligate the blunt ends of any DNA molecule and cut with specific restriction enzymes and produce DNA fragments with sticky ends. Adapters contain performed sticky or cohesive ends. These are useful to ligation of DNA fragments with blunt ends.
26. Write a note on DNA ligase?
Ans: The cut DNA fragments are covalently joined with plasmids by DNA ligases. These enzymes are originally isolated from viruses. These are also occur in E.coli and eukaryotic cells. These enzymes joins the DNA fragments by forming a phosphodiester bond between phosphate group of 5 carbon of one deoxy ribose with hydroxyl group 3 carbon of another deoxy ribose.
27.What is dane particle?
Ans: Hepatitis B virus consisting of size is 42 nano meters, which is called "dane particle", this particle consisting of "core", in this core contain viral genome(DNA) which is surrounded by phospholipid envelope.
28. Explain the outline of r-DNA technology?
Ans: The basic principles of r-DNA technology are reasonably simple, which involves the following steps.
- Generation of DNA fragments and selection of desired piece of DNA.
- Insertion of this selected DNA in to cloning vector to produce recombinant DNA or chimeric DNA.
- Multiplication and selection of clones from recombinant molecules.
Ans: Gel electrophoresis is a routinely used analytical technique for the separation or purification specific DNA fragments. The gel is composed of either polyacrylamide or agarose. Polyacrylamide gel electrophoresis is used for the separation of large DNA fragments ranging in size from 100 base pairs to 20 kb base pairs. Gel electrophoresis is also used for the separation of RNA molecules.
30. What are plasmids? Give the classification of Plasmids?
Ans: plasmids are extra chromosomal, double stranded, circular self replicating DNA molecules
31.Explain in details about PBR 322 Plasmid?
Ans: PBR 322 is a plasmid discovered by Boliver and Rodriguez they given the numerical number is 322. Due to this reason this is called by PBR 322 Plassmid.
32.Define restriction endonucleases?
Ans: Restriction endonuclease or restrictase is an enzyme that cleaves DNA into fragments at or near specific recognition sites within molecules known as restriction sites. Restriction enzymes are one class of the broader endonuclease group of enzymes.
33.Write a note on DNA modifying enzymes?
Ans: The cut DNA fragments are covalently joined with plasmids by DNA ligases. These enzymes are originally isolated from viruses. These are also occur in E.coli and eukaryotic cells. These enzymes joins the DNA fragments by forming a phosphodiester bond between phosphate group of 5 carbon of one deoxy ribose with hydroxyl group 3 carbon of another deoxy ribose.
34. Explain in details suitable host cells for r-DNA technology?
Ans: In generally microorganisms are preferred as host cells, because they are multiply faster when compared to cells of plants and animals.
35. What is Polymerase chain reaction?
Ans: The polymerase chain reaction is a laboratory technique for generating large quantities of a specified DNA. Obiviously, PCR is a cell-free amplification technique for synthesizing multiple identical copies of any DNA of interest. Developed in 1994 by karry Mullis PCR is now considered as a basic tool for the molecular biologist.
36.What is the principle involved in the polymerase chain reaction?
Ans: The double-stranded DNA of interest is denatured to separate into two individual strands. EAch strand is then allowed to hybridize with a primer. the primer-template duplex is used for DNA synthesis. These three steps-denaturation, renaturation and synthesis are repeated again and again to generate multiple forms of target DNA.
37. What is annealing ?
Ans: As the temperature of the mixture is slowly cooled to about 55°C, the primers base pair with the complementary regions flanking target DNA strands. This process is called renaturation or annealing. High concentration of primer ensures annealing between each DNA strand and the primer rather than the two strands of DNA.
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