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Introduction to Pharmaceutical Biotechnology, Volume 2 (Second Edition)
the capacity they provide for the development of treatments that precisely address the underlying molecular causes of disease [120].
1.15.6 Enzyme biomarkers in disease diagnosis
Enzymes are frequently used in various medical diseasesdiagnostic procedures because they serve as measurable indicators of biological processes or disease states. Cardiac troponins are enzymes that are released into the bloodstream in reaction to injury to heart muscle cells (myocardial infarction), and they are crucial markers for diagnosing heart attacks. High levels of cardiac troponin in the blood can be used to diagnose and treat acute myocardial damage [121].
1.15.7 Pharmacogenomics and enzyme variability
Pharmacogenomics studies how an individuals genetic makeup affects their response to drugs. The metabolism and effectiveness of medications are profoundly affected by enzyme variability. The CYP enzyme family metabolizes many different medications. Variations in the CYP genes may affect the capacity to metabolize certain drugs. Differences in CYP might cause slower medication metabolism, which can increase blood drug levels and increase the risk of adverse drug reactions in some persons. Knowing a patients CYP enzyme prole could help clinicians choose the best treatment and dose for that individual [122].
1.15.8 Enzyme-based therapies for personalized treatment
Those genetically predisposed to lack certain enzymes may benet from ERT, which entails giving them therapeutic enzymes. The severity of enzyme decits in lysosomal storage diseases can vary widely between different mutations. Determining the precise enzyme deciency of each patient is crucial for providing successful, tailored enzyme replacement medication. This customized approach guarantees that each patient receives the most effective enzyme and dosage possible [123].

1.16 Enzymes in bioremediation

Bioremediation is a sustainable method for restoring contaminated environments. Enzymes are essential to improving bioremediation systems due to their catalytic characteristics. When enzymes are added to wastewater, organic molecules, and contaminants degrade more quickly, producing less harmful by-products or becom­ing harmless altogether. Lipases are enzymes that metabolize fats and oils into their constituent fatty acids and glycerol. Lipases could be used to hydrolyze fats and oils in domestic and industrial wastewater, decreasing the risk of clogged pipes and easing some harmful impacts of these discharges on the environment [124]. Enzymes can catalyze the conversion of hazardous compounds, hydrocarbons, and other organic contaminants into less toxic molecules. Enzymes called alkane hydroxylases hydrolyze alkanes, making up most of crude oil. When applied to oil-stained areas, these enzymes speed up the natural recovery process. The breakdown of the hydrocarbons makes this possible [125]. Soil bioremediation is aided by enzymes,
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Introduction to Pharmaceutical Biotechnology, Volume 2 (Second Edition)
which speed up the breakdown of organic molecules and the removal of contam­inants. Phytoremediation is a method that uses plant roots and the microorganisms that live in them to detoxify soil from toxins. The enzyme phytase can remediate polluted soils by promoting plant phosphorus absorption and mobilization from organic molecules [126].

1.17 Enzymes in agriculture and crop production

Enzymes are natural catalysts that help plants speed up their metabolism. The many benets of employing them in agriculture include improved nutrient absorption, insect and disease resistance, post-harvest convenience, and longer shelf life [127]. The availability of enzymes and nutrient absorption may improve plantsresistance, development, and growth. Phosphatases are enzymes that break down organic matter in the soil, freeing up phosphate for plant uptake. By increasing the amount of phosphate available to the plant, phosphatases improve germination rates, root growth, and general plant health. This is especially useful because plants may have problems receiving adequate phosphate from the soil in low-phosphate
Figure 1.8. (A) Conceptual diagram outlining the role of microbial enzymes in the catalytic degradation of environmental pollutants to non-toxic metabolites. (B) Schematic representation of the microalgal biomass production cycle illustrating nutrient recycling and bioresource applications.
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Introduction to Pharmaceutical Biotechnology, Volume 2 (Second Edition)
environments [128]. Reduced reliance on chemical pesticides results from cutting­edge strategies that use enzymes to shield crops against pests, illnesses, and environmental stressors. Chitinases are digestive enzymes that break down insect and arthropod exoskeletons made of the protein chitin. Chitin-eating insects and pests have no chance against plants that express the chitinase gene. This enzymatic method can help protect crops from pests while decreasing the need for chemical pesticides [129]. Post-harvest handling and storage with the help of enzymes could help keep harvested crops fresh and usable for longer. Polygalacturonate is an enzyme that helps ripen fruit. Manipulating this enzymes activity may lengthen or reduce the post-harvest storage duration. Inhibiting polygalacturonate activity may extend the freshness of some fruits, such as tomatoes, throughout storage and shipping [130].

1.18 Enzymes in waste management

Enzymes are necessary for biodegradation and energy generation from a wide variety of waste, making waste management an essential aspect of environmental sustainability. Compost, rich in nutrients that can be used to improve soil, is produced more quickly by adding enzymes to the natural decomposition process of organic waste. Plant cell walls contain cellulose, an essential structural component that can be degraded by enzymes known as cellulases. The composting process can be sped up with cellulases, which help decompose organic waste such as food scraps, yard debris, and leaves [131]. High-quality compost is produced by this method, and it can be utilized as a fertilizer in sustainable farming by increasing soil fertility with natural means [132]. Enzymes are used to break down plastic waste into simpler molecules to lessen plastic pollution and its negative impacts on the environment. PETase can break down polyethylene terephthalate (PET), a standard plastic. We now know which bacteria manufacture PETase naturally, and thanks to genetic engineer­ing, we can make the enzyme more effective. An enzymatic method shows promise for efciently recycling PET plastic, which could help keep plastic trash out of landlls and waterways [133]. Enzymes are used in waste-to-energy conversion systems to convert organic waste into biogas and biofuels. With the help of bacteria and enzymes, organic waste such as food scraps and agricultural leftovers can be broken down in anaerobic digestion even when oxygen is unavailable. Biogas, which is primarily methane, is created by this method and represents a renewable energy source. Using biogas to fuel vehicles, heat homes, or generate electricity helps minimize emissions of greenhouse gases caused by waste breakdown (gure 1.8)[134].

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Introduction to Pharmaceutical Biotechnology, Volume 2
(Second Edition)
Enzymes, proteins and bioinformatics
Ahmed Al-Harrasi, Saurabh Bhatia and Ajmal Khan
Chapter 2
Technologies and procedures involved in
enzyme production

2.1 Introduction

Enzyme production has become a signicant area in the modern biotechnology industry. With advancements in traditional enzyme production methodologies, different industrial sectors can now easily exploit new applications offering novel activities and/or improved stability. Enzymes have been used by human beings since ancient times, either in the form of vegetables rich in enzymes, or in the form of microorganisms. These microorganisms have been used for many purposes, such as in brewing, and the baking industry. In the late nineteenth century numerous biological conversions were attributed to the action of enzymes. Isolation was rst introduced by Payen and Persoz in 1833 [1]. They isolated an enzyme complex from malt, which they named diastase, that converts gelatinized starch into sugars, primarily maltose. In 1874, the history of modern enzyme production really began when Christian Hansen rst synthesized rennet by isolating it from dried calves stomachs with saline solution [2]. This was the rst enzymatic preparation of relatively high purity used for commercial purposes. The Christian Hansen Company is still developing rennet today, in an almost identical way. In 1896, the utilization of the mold fungus koji in the production of certain foodstuffs and avor additives based on soya protein and fermented beverages was the basis on which the Japanese researcher Takamine established a fermentation process for the industrial production of fungal amylase.
This process involves culturing of Aspergillus oryzae on moist rice or wheat bran, and the product is called Takadiastasewhich is currently used as a pharmaceutical agent to cure digestive disorders [3]. The market value of the industrial enzymes market was estimated as approximately $2 billion, and has increased at an average annual rate of 3%–5% in the last decade. Several enzyme production based
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