For industrial applications, enzymes are modified to possess certain characters. The enzyme should have a long life.
The enzyme should be able to use substrate supplied in the industry adequately
The enzyme should be capable of working under extreme temperature, pH, pressure, and concentration even if different from those present inside the cell.
Methods for Modification of Enzymes
There are two methods for modifying enzymes by mutagenesis. One is Random mutagenesis, and another one is site-directed mutagenesis.
Random mutagenesis Mutation is concerned with the alteration in the sequences of the genetic material and this random mutagenesis is commonly observed at the random locations of the genetic material. Some of the examples of random mutations are transition and insertions. Deletion of the genes also belongs to this mutagenesis, in this type, there is the loss of a segment of some genes. Some regions are impaired during the synapsis and these form the loop at the time of synapsis. In the final product, a heterozygotic phenotype is formed. Duplication is also an example of mutagenesis, in which there is a duplication of the gene occurs, a perfect example for the duplication is the addition of the 16 A segment in the X chromosome in the fruit fly that leads into the bar-shaped eye. There is the rotation of the segment of the chromosome at the degree of 180, this kind of mutagenesis is termed as inversion. In this, it inhibits the crossing over, so it is known as a cross-over repressor. The scientist Muller used the CIB method that is used for the identification of the mutation that involves the inversion.
Error-prone PCR: This type of polymerase chain reaction works on the truth that the enzyme which is important for a polymerase chain reaction that is Taq DNA polymerase does not consist of 3â€™ to 5′ exonuclease activity due to which there are chances of a mistake rate is 0.002percent per nucleotide at each of the replication. Chemical mutagenesis- This kind of mutagenesis is caused due to usage of the chemical matter that may cause mutagenesis. An example of chemical mutagenesis is sodium bisulfate. All the chemical that is involved in this mutagenesis can cause transversion.
Site-directed mutagenesis- It is a typical point mutation that is found in a particular known area. This mutation is site-specific and gives rise to wild type and mutated DNA.
Post-Translational Modification of Enzymes
Post-translational modification refers to a change in the protein or enzyme structure after translation. Several covalent modifications are done in the case of enzymes which includes methylation in which methyl group is added to enzyme molecule, acetylation in which acetyl group is added to the modified enzyme, and glycosylation in which sugar moiety is added. Let us take the example of Zymogens, which are an inactive form of an enzyme, these enzymes undergo covalent modification and get converted into an active form such as in the case of trypsinogen, which is an inactive enzyme, this trypsinogen undergoes chemical modification and produces active trypsin. Things modified in enzymes: Binding refers to the interaction of proteins with their surroundings. In this modification number of binding points is increased to bind a molecule with high affinity. Catalysis To improve the catalytic rate, the binding to the transition state is increased. Increasing stability To increase the stability of the enzyme, several chemicals and bonds are introduced in the enzyme, such as for the stability of T4 lysozyme, S-S bonds are introduced.
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