In recent years, there has been significant progress in establishing molecular genetics tools for extreme thermophiles, allowing the use of these microorganisms as metabolic engineering platforms. Thermophilic enzymes, which are highly desirable due to their efficient catalytic activity at high temperatures, have gained interest in industrial applications. However, most enzymes exhibit inferior performance. An ancestral sequence-structure-molecule dynamics (ASSMD) strategy was designed to unveil molecular insights into extinct ancestral enzymes in the evolutionary past.
Thermophilic polyester hydrolases (PES-H) have recently enabled biocatalytic recycling of mass-produced synthetic polyester polyethylene terephthalate (PET), showing that by replacing traditional chemical methods with enzyme-driven reactions, complex products can be produced efficiently. Enzyme engineering is a growing field, with thermophilic bacteria growing at higher rates compared to mesophilic counterparts and having lower cooling requirements and less contamination.
Enzymes from microorganisms growing at elevated temperatures are termed thermozymes and considered ideal candidates for catalytic processes operating at high temperatures. It is becoming increasingly possible to improve the thermostability of mesophilic enzymes through protein engineering or techniques such as enzyme technology tools.
Innovation in thermophilic synthetic biology is essential, as engineering mesophilic enzymes with desired higher thermostability features allows for a fusion of both higher enzymatic activity and thermostability. Thermophilic microorganisms from hot springs are a valuable source of stable enzymes for biotechnological applications, and investing time and effort into developing a thermophilic synthetic biology direction is beneficial.
In conclusion, the development of thermophilic synthetic biology is crucial for the development of efficient and eco-friendly biocatalysts for various industrial applications.
| Article | Description | Site |
|---|---|---|
| Progresses and challenges of engineering thermophilic … | In particular, the absence of characterized genetic parts and thermostable enzymes represents a significant gap in the field of metabolic engineering, as these elements are crucial for advancing this area of research. It is notable that… | www.frontiersin.org |
| Thermophiles and the applications of their enzymes as … | A review of recent discoveries of thermophilic enzymes and their current and potential applications in industry will be presented.by J. Atalah · 2019 · Cited by 247 | www.sciencedirect.com |
| Potential and utilization of thermophiles and thermostable … | By P. Turner, 2007. Cited 759 times. This article discusses existing and potential applications of thermophiles and thermostable enzymes, with a particular focus on the conversion of carbohydrate-containing raw materials. | pmc.ncbi.nlm.nih.gov |
📹 Engineering thermostable and cryostable Enzyme and Site directed mutagenesis
Subject:Biophysics Paper:Molecular ezymology and protein engineering.
Can scientists create enzymes?
Bioengineered enzyme can produce synthetic genetic material, advancing development of new therapeutic options. A research team led by the University of California, Irvine has engineered an efficient new enzyme that can produce a synthetic genetic material called threose nucleic acid.
Why are scientists interested in thermophilic bacteria?
The ability of microorganisms to survive under harsh conditions has prompted researchers to study these organisms to better understand their characteristics and eventually utilize them in various applications. Further insight into thermophilic microorganisms has been highlighted through this review article as thermophiles possess enumerable properties suitable for biotechnological and commercial applications.
Biotechnological applications of thermophiles. Thermophiles have shown tremendous promise in terms of their applications in modern biotechnology. Some of the high end applications of these thermophiles have been elucidated below (Fig. 1 ).
Various applications of thermophilic microorganisms.
What are thermophilic enzymes used for?
Thermophilic enzymes are a versatile and efficient solution for various industrial processes, including chemical, food, pharmaceutical, paper, and textile industries. They are known for their exceptional operational stability at high temperatures and denaturant tolerance, making them ideal for converting biomass into target products. Biorefineries use microbial cells and their enzymes to convert biomass into target products, allowing for easy mixing, better substrate solubility, high mass transfer rate, and reduced contamination risk.
Thermophiles have been proposed as sources of industrially relevant thermostable enzymes, with a focus on the conversion of carbohydrate-containing raw materials. Their importance in biorefineries is explained through examples of lignocellulose and starch conversions to desired products. Strategies that enhance the thermostablity of enzymes both in vivo and in vitro are also assessed.
Thermophiles and microorganisms have been studied extensively, starting in the 1960s with the pioneering work of Brock and his colleagues. Microorganisms are divided into three main groups: psychrophiles (below 20°C), mesophiles (moderate temperatures), and thermophiles (high temperatures, above 55°C). Thermophilic bacteria typically grow below the hyperthermophilic boundary, while hyperthermophilic species are dominated by the Archaea.
The development of molecular biology techniques, allowing genetic analysis and gene transfer for recombinant production, led to increased activities in the field of thermostable enzymes during the 1990s. This led to the isolation of microbes from thermal environments to access enzymes that could significantly increase the window for enzymatic bioprocess operations.
Industrial enzymes are also being explored for their potential use in technical products and processes, often on a large scale. These enzymes offer robust catalyst alternatives that can withstand harsh industrial processing conditions.
Can an enzyme be engineered?
Enzyme engineering is a powerful tool enabling these approaches primarily based on the optimization of amino acid sequence. Currently, it is still technically difficult to design an artificial enzyme in vitro. Hence, a natural enzyme close to the final target form has to be selected first for engineering.
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How can we grow thermophiles in the laboratory?
Thermophile bacteria is growing planted into medium of selective starch agar 1% (10 g/l starch and 15 g/l bacto agar) and incubated at temperature 50°C for 24 hours. Bacteria that was grown on selective medium was dripped with iodine solution.
How are thermophiles used in industry?
Thermophilic microalgae and cyanobacteria have gained attention for their biotechnological and industrial potential, including thermostable enzymes, high-value molecules like phycobiliproteins and carotenoids, biopolymers, biofuels, wastewater treatment, and bioremediation. Extremophilic microorganisms play a crucial role in understanding the origin and evolution of life on Earth, and their ability to survive at extreme temperatures, pressures, salinity, and pH values has provided insights into biodiversity, adaptation mechanisms, and the evolutionary history of life on Earth.
Thermophiles and hyperthermophiles exhibit complex genetic and physiological changes that evolved as a response to temperature-related stress. Protection mechanisms rely on factors such as altering cell morphology, activating detoxification enzymes to counteract oxidative stress, overexpressing heat shock proteins, and the presence of chemically stable lipids within membranes. Structural studies reveal that amino acid substitutions found on the surface regions of thermophilic proteins are responsible for an increased number of intermolecular salt bridges and hydrogen bonds that stabilize secondary and tertiary structures.
The biotechnological potential of thermophilic microorganisms has been promoted as an alternative to mesophilic ones in industrial processes. To be effective, biotechnological processes should allow large-scale production of bio-based products by cost-effective procedures carried out in sterile conditions, minimizing the risk of contamination. Temperature is a critical parameter in industrial processes that can influence the growth rate of microorganisms, catalytic activity of enzymes, and bioactivity of the molecules involved. High temperatures are widely used in various industrial sectors for purposes such as food sterilization and extraction procedures to improve extraction yields.
This review aims to explore recent advancements in using thermophilic microbiomes, algae, and bacteria for applications in bioenergy, bioremediation, and the synthesis of valuable bioproducts.
What are the enzymes used in engineering?
1. 4. 3 Protein DesignEnzymesMutation MethodReferencesThrombin-thrombomodulin fusion proteinsSite-directed mutagenesisLipaseDirected evolutionIr(Me)-myoglobin and generated mutantsIntroducing new chemical activitiesBacillus circulans xylanaseSite-directed (strategically selecting the mutation sites)
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Why are scientists so interested in thermophilic bacteria?
Thermophilic microbes are important sources of thermozymes. The stability of thermozymes at high temperatures is often utilized in harsh industrial processes and makes them a valuable resource for biotechnological applications.
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What are the techniques of enzyme engineering?
The general approaches in enzyme engineering include rational designing, semi-rational design, DNA shuffling, structure base design and directed (molecular) evolution, random mutagenesis, cell surface display technique, etc.
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Can we genetically engineer enzymes?
Food enzyme modification is developed by advanced genetic techniques. Genetically modified food enzymes can exhibit improved catalytic efficiency, stability, substrate specificity. Most genetically modified food enzymes are designed to be applied in food processing involving carbohydrates and lipids.
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Can enzymes be artificial?
To rival natural enzymes, various artificial enzymes have been developed over the last decades. Since supramolecular interactions play important roles in both substrate recognition and the process of enzymatic catalysis, designing artificial enzymes using supramolecular strategies is undoubtedly significant.
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📹 M-25. Engineering thermostable and cryostable Enzyme and Site directed mutagenesis
… these thermophilic enzymes in mesophilic hosts as we normally do we will look at that during further slides these enzymes can …
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