Enzyme Technology
Enzyme Technology
In living things, the enzyme serves as a catalyst, controlling the speed at which chemical processes take place without undergoing any changes. Since the dawn of civilization, natural enzymes have been employed in the creation of goods like leather, indigo, and linen as well as foods like cheese, sourdough, beer, wine, and vinegar.
All of these activities depended on either enzymes created by microorganisms that were developing on their own or enzymes found in additional preparations like papaya fruit or calves’ rumen. Enzyme technology is the study of industrial enzymes and their applications. For the benefit of humanity, enzyme technology generally entails the manufacture, separation, purification, and application of enzymes (in soluble or immobilized form).
Enzyme technology also includes the use of protein engineering and recombinant DNA technologies to create enzymes that are more effective and useful. Thus, the term “enzyme technology” today primarily refers to the use of modifying an enzyme’s structure (and thereby its function) or the catalytic activity of isolated enzymes in order to produce new metabolites, to enable the occurrence of new (catalyzed) pathways for reactions, or to convert some specific compounds into others (biotransformation).
Objective
To create brand-new, environmentally friendly services, processes, and goods to satisfy human needs, or to enhance production methods for current goods using fresh biomass and raw materials.
Value of Enzyme Technology
Enzymes are used in numerous processes. These include the production of food, it’s processing, and preservation, the creation of washing powders, the production of textiles, the leather, and paper industries, the development of medicinal applications, the enhancement of the environment, and scientific research. According to contemporary estimates, the bulk of industrially generated enzymes is helpful in processes involving foods (45%), detergents (35%), textiles (10%), and leather (3%), among other things.
Products made using enzyme technology can be employed as chemicals, pharmaceuticals, fuel, food additives, or agricultural enhancements. A few characteristics that can be altered or improved by genetic engineering include the yield and kinetics of the enzyme, the ease of the subsequent process, and various safety concerns. In order to develop safe, high-production microorganisms, enzymes from dangerous or forbidden germs as well as from slowly growing or scant plant or animal tissue can be cloned. In the future, enzymes might be modified to better fit industrial operations.
We shall pursue ever-higher objectives for enzyme engineering by complementing full-gene random mutagenesis with strategies that direct mutation to specific protein regions or use recombination to begin major sequence alterations.
Enzyme technology applications
Enzymes are used as biocatalysts in the field of enzyme technology, a subsection of biotechnology, to produce both bulk and high-value products, addressing needs in the production of food (such as bread, cheese, beer, and vinegar), fine chemicals (such as amino acids, vitamins), and pharmaceuticals. Enzymes are also utilized for analytical and diagnostic applications, as well as for services like cleaning and environmental procedures.
Enzyme technology advances
- In the natural world and through strain selection, new enzymes are being sought after.
- As many different applications for established commercial enzymes as are imaginable exist.
- Genetic engineering is used to create brand-new enzymes.
- Using the “know-how” developed from enzymology, new organic catalysts are being created and manufactured.
- Enzyme systems are becoming more sophisticated.
Upcoming Applications
- Enzymes being used as electrocatalysts (specific biosensors)
- Enzymes as analytical tools for analyzing particular substances and for the regeneration of particular metabolites
- Utilization of enzymes in the synthesis of large-scale organic compounds and the creation of perfumes and cosmetics
- Utilizing enzymes to create chemicals that give food its tastes and aromas using enzymes as a technique for the removal of pesticide traces
- Monitoring hazardous chemical levels in food and water using enzymes
Applications of enzyme technology in biomedicine will include
- The creation of fresh anti-microbial substances
- replacement of enzymes
- Cancer therapy with enzymes
- Applications in dermatology and enzyme graft
- Enzymes as precursor molecule activators
- Utilizing enzyme technology to prevent dental cavities
Submerged Fermentation
Submerged Fermentation
The biological process of turning complicated substrates into simple chemicals by a variety of microorganisms, including bacteria and fungus, is known as fermentation. They emit a number of other substances during this metabolic breakdown in addition to the typical byproducts of fermentation, such as carbon dioxide and alcohol. These extra substances are referred to as secondary metabolites. Many different antibiotics, peptides, enzymes, and growth hormones are examples of secondary metabolites. The advancement of processes like Solid State Fermentation (SSF) and Submerged Fermentation (SmF) has enabled the manufacture of bioactive chemicals at an industrial scale.
Submerged fermentation
In the process of producing biomolecules known as “submerged fermentation,” enzymes and other reactive substances are immersed in a liquid such as alcohol, oil, or nutritional broth.
Liquid fermentation (LF) and submerged fermentation (SmF) both use freely flowing liquid substrates such as broths and molasses. The method is employed for many different things, mostly in industrial manufacturing.
Submerged Fermentation Principle
When a microbe ferments submerged, it grows as a suspension in a liquid medium that contains a variety of nutrients that are either dissolved or suspended as particulate solids in many commercial media. A method known as submerged fermentation involves the growth of microorganisms in a liquid broth. This liquid broth produces industrial enzymes, antibiotics, and other goods in addition to providing nutrients. The procedure entails adding a particular microorganism, such as fungus, to a tiny, closed flask filled with nutrient-rich broth. Additionally, the procedure needs a lot of oxygen. When the microorganisms in the broth interact with the nutrients, breaking them down, enzymes are then produced. The fermentation broth is where the bioactive substances are released.
Techniques for Submerged Fermentation
Batch-fed fermentation and continuous fermentation are the two most often used techniques for submerged fermentation. In batch-fed fermentation, the culture is supplemented with sterile growth ingredients. As it happens during the growth of biomass in the fermenter, it is most prevalent in the bio-industry. It is normally highly concentrated to prevent dilution and aids in increasing cell density in the bioreactor. By supplying nutrients, the culture’s development rate is maintained, and the risk of overflow metabolism is decreased. The fermentation process is carried out continuously in an open system. The converted nutrient solution is then slowly and continually collected from the bioreactor while sterilized liquid nutrients are introduced.
Examples of Submerged Fermentation Applications
- The main use of SmF is the liquid-form extraction of secondary metabolites.
- Enzymes originating from microorganisms are typically produced by submerged liquid fermentations.
Submerged fermentation benefits
- The benefits of submerged fermentation techniques are quick turnaround, low cost, and high yield.
- Product purification is simpler.
- Because fermentation can be more easily controlled in liquid culture, fermentation times can be cut down significantly.
- Similar to how solid-state technologies need more labor, submerged culture can increase the production of several secondary metabolites while lowering production costs.
Submerged fermentation’s drawbacks
- Recent studies have shown that SSF has a significant impact on productivity, resulting in higher yields and better product qualities than SmF.
- Ineffective volumetric productivity
- Comparatively lower product concentration
- Increased wastewater production
- Sophisticated fermentation apparatuses
Ideonella sakaiensis (plastic-eating bacteria) – An Overview
Ideonella sakaiensis (plastic-eating bacteria) - An Overview
A new species of the genus Ideonella called Ideonella sakaiensis is a rod-shaped, gram-negative aerobic bacteria. It is also known as a bacterium that consumes plastic. Plastic can serve as a key source of carbon and energy for growth for bacteria that consume it.
With Leo Baekeland’s discovery of Bakelite in 1907, the era of plastics began. The creation of a wide variety of synthetic polymers has led to their widespread use and necessity in both everyday life and, tragically, the environment.
In 2015, there was 5,000 Mt of plastic garbage in landfills and other places. That amount could increase to 12,000 Mt by 2050. Numerous hydrolytic enzymes can hydrolyze the ester bonds that bind the monomers of poly (ethylene terephthalate). Because there are no known enzymes that may directly break their C-C bonds, PET is theoretically more prone to spontaneous deterioration than polyolefin. The genome of I. sakaiensis was sequenced, and this led to the discovery of an enzyme that hydrolyzes PET. The hydrolysis products are discharged into the aqueous environment, and the surface of an amorphous PET film develops pitting that resembles craters. The I. sakaiensis enzyme was given the name PET hydrolase because, when compared to other known PHEs, it displays the highest catalytic preference for PET at ambient temperature (PETase). This bacterium converts PET into CO2 and water, using it as a main source of carbon and energy.
Ideonella sakaiensis Discovery
A group of scientists led by Kohei Oda of Kyoto Institute of Technology and Kenji Miyamoto of Keio University collected 250 samples from a PET-contaminated environment, including sediment, soil, wastewater, and activated sludge close to a plastic bottle recycling facility. PET was introduced into this ecosystem. Using these samples, they searched for microbes that could grow preferentially on low-crystallinity (1.9%) PET film. After being cultivated in a single sediment sample, distinct microbial consortia formed on the PET film, resulting in morphological changes. Ideonella sakaiensis is a species that belongs to the Ideonella genus. The bacteria was given the name Sakai after the Japanese city where it was found.
A collection of microbes in the soil sample, which also contained protozoa and cells that resembled yeast, led to the initial isolation of the bacterium. On the film and in the culture fluid, appendages connecting the cells were visible. There were shorter appendages that may have assisted in delivering secreted enzymes to the film between the cells and the film. The PET film had undergone substantial deterioration and was nearly destroyed after six weeks at 30°C.
Ideonella sakaiensis Classification
Ideonella sakaiensis, a member of the genus Ideonella of the family Comamonadaceae, was discovered to degrade plastic in response to the hunt for a biological system that could degrade plastic waste. In the Betaproteobacteria 16S rRNA phylogeny, members of the Comamonadaceae family are found together in a phylogenetic cluster.
| Domain | Bacteria |
|---|---|
| Phylum | Pseudomonadota |
| Class | Betaproteobacteria |
| Order | Burkholderiales |
| Family | Comamonadaceae |
| Genus | Ideonella |
| Species | Ideonella sakaiensis |
Habitat of Ideonella sakaiensis
- Ideonella sakaiensis can be found in a variety of both natural and artificial environments, and they don’t care if a place is clean or dirty.
- Soil, fresh and groundwater, activated sludge, and water used in industrial processes are a few examples of these ecosystems.
- The physical and physiological traits of the distinct features of the genera vary widely within the species themselves as a result of the large variety of habitats.
Mode of action of Ideonella sakaiensis
PETase enzyme
- PET is broken down by the bacterial enzyme PET hydrolase, or PETase, creating mono (2-hydroxyethyl) terephthalic acid (MHET), a heterodimer consisting of terephthalic acid (TPA) and ethylene glycol (EG).
- PET’s ester bonds are hydrolyzed by I. sakaiensis PETase.
- MHET hydrolase is a different enzyme that is utilized since I. sakaiensis and many other bacteria cannot use TPA but can readily absorb and use ethylene glycol among the products.
- In other words, both chemicals derived from the PET are used by the cell to produce energy and for metabolic functions.
- This system, in particular, ranks among the most effective PET hydrolases known to exist at room temperature in terms of PET breakdown efficiency.
- As carbon is assimilated, it may eventually mineralize into carbon dioxide and be released into the atmosphere.
MHET hydrolase
- The Ideonella sakaiensis genus has another tannase-family enzyme involved in PET metabolism.
- This enzyme hydrolyzes 2-hydroxyethyl terephthalic acid (MHET), the main PET hydrolysis product of PETase, into TPA and EG.
- Due to its strong activity and excellent specificity for mono-(2-hydroxyethyl) terephthalate (MHET), this enzyme was given the name MHET hydrolase (MHETase), but it has a modest hydrolytic effect on bis-(2-hydroxyethyl) TPA (BHET) and a number of other substrates.
- Its composition comprises a lid domain and a hydrolase domain was revealed by the crystal structure.
Antimicrobial Resistance (AMR) and Antibacterial Resistance (ABR)
Antimicrobial Resistance (AMR) and Antibacterial Resistance (ABR)
A major public health concern is the development of antimicrobial resistance (AMR) in pathogenic bacteria, which is likely to lead to worsened sickness, higher death, and increased treatment costs with constrained treatment alternatives. The AMR Review from 2014 predicted that if this issue is not addressed, there will be much more deaths than the expected 10 million per year by the year 2050.
Antimicrobial resistance is the capacity of microorganisms to fend off the effects of drugs that, in the past, would have killed them or prevented their growth. Microbes can continue to grow even after being exposed to antimicrobial agents thanks to the development of antimicrobial resistance. The length of time it takes to recover from an illness, the severity of the sickness, the amount of treatment required, and the potential fatality risk all rise if pathogens acquire resistance.
Studies and observations of AMR in bacteria and fungus are more frequent. A small number of parasites have also evolved resistance to their available treatments. Antibacterial resistance (ABR) is the term used when bacteria become resistant to “antibiotics”. Antifungal resistance (AFR) is the term used to describe a fungus’ development of resistance to “antifungals”. The development of resistance by viruses and helminths against their respective treatments is referred to as “antiviral resistance” and “anthelmintic resistance,” respectively.
The most significant form of AMR is antibacterial resistance (ABR), which has been linked to major infections through the development of resistance in many pathogenic bacterial species. Although it is less common than bacterial resistance, resistance in fungi, viruses, and parasites is also becoming more often reported.
Four AMR mechanisms in general cause the development of AMR
- Possibility of altering or inactivating the medication
- Decrease in medication affinity or absorption
- An increase in drug efflux
- Changing the cellular components that are the drug’s target spot
These mechanisms either originate as a result of gene insertions or mutations that affect any of these systems. Microbes may only be resistant to one type of antibiotic or they may be resistant to several. If the resistance is limited to one class of antimicrobials that are structurally and mechanistically similar or identical, it is simply referred to as that class of resistance. For instance, a bacterium is simply referred to as a “penicillin-resistant bacteria” if it is resistant to penicillin and its derivatives.
Resistance to numerous antimicrobials with distinct structures and modes of action is referred to as multi-drug resistance (MDR). A popular definition is that an antibiotic is an MDR if it is resistant to at least “one antimicrobial in three or more categories” of structurally unrelated antimicrobials. MDR pathogens are known as the “SUPER BUGS.” Additional categories for MDR include extensively drug-resistant (XDR) and universally drug-resistant (PDR). Antibiotics from no more than two antibiotic classes that are structurally unrelated will be effective against XDR infections. The PDR pathogens will be immune to all antimicrobials.
AMR has been listed as one of the top 10 global public health hazards by the World Health Organization (WHO). In a paper titled “Global priority list of antibiotic-resistant bacteria to guide research, discovery, and Development of novel antibiotics,” the WHO published a list of pathogens in 2017. In its 2019 AR Threat Report, the US Centers for Disease Control and Prevention (CDC) published a list of resistant bacteria and fungi. In addition to pathogens of concern in the US, this list also contains microorganisms from the WHO’s 2017 list.