Thermophilic Research Articles

Structural insights into thermophilic chaperonin complexes

Group I chaperonins are dual heptamer protein complexes that play significant roles in protein homeostasis. The structure and function of the Escherichia coli chaperonin are well characterized. However, the dynamic properties of chaperonins, such as large ATPase-dependent conformational changes by binding of lid-like co-chaperonin GroES, have made structural analyses challenging, and our understanding of these changes during the turnover of chaperonin complex formation is limited. In this study, we used single-particle cryogenic electron microscopy to investigate the structures of GroES-bound chaperonin complexes from the thermophilic hydrogen-oxidizing bacteria Hydrogenophilus thermoluteolus and Hydrogenobacter thermophilus in the presence of ATP and AMP-PNP. We captured the structure of an intermediate state chaperonin complex, designated as an asymmetric football-shaped complex, and performed analyses to decipher the dynamic structural variations. Our structural analyses of inter- and intra-subunit communications revealed a unique mechanism of complex formation through the binding of a second GroES to a bullet-shaped complex.

Abstract 1225 Bioplastic degradation by a polyhydroxybutyrate depolymerase from a thermophilic soil bacterium

Abstract 1225 Bioplastic degradation by a polyhydroxybutyrate depolymerase from a thermophilic soil bacterium

The protein-only RNase Ps, endonucleases that cleave pre-tRNA: Biological relevance, molecular architectures, substrate recognition and specificity, and protein interactomes.

Protein-only RNase P (PRORP) is an essential enzyme responsible for the 5' maturation of precursor tRNAs (pre-tRNAs). PRORPs are classified into three categories with unique molecular architectures, although all three classes of PRORPs share a mechanism and have similar active sites. Single subunit PRORPs, like those found in plants, have multiple isoforms with different localizations, substrate specificities, and temperature sensitivities. Most recently, Arabidopsis thaliana PRORP2 was shown to interact with TRM1A and B, highlighting a new potential role between these enzymes. Work with At PRORPs led to the development of a ribonuclease that is being used to protect against plant viruses. The mitochondrial RNase P complex, found in metazoans, consists of PRORP, TRMT10C, and SDR5C1, and has also been shown to have substrate specificity, although the cause is unknown. Mutations in mitochondrial tRNA and mitochondrial RNase P have been linked to human disease, highlighting the need to continue understanding this complex. The last class of PRORPs, homologs of Aquifex RNase P (HARPs), is found in thermophilic archaea and bacteria. This most recently discovered type of PRORP forms a large homo-oligomer complex. Although numerous structures of HARPs have been published, it is still unclear how HARPs bind pre-tRNAs and in what ratio. There is also little investigation into the substrate specificity and ideal conditions for HARPs. Moving forward, further work is required to fully characterize each of the three classes of PRORP, the pre-tRNA binding recognition mechanism, the rules of substrate specificity, and how these three distinct classes of PRORP evolved. This article is categorized under: RNA Structure and Dynamics > RNA Structure, Dynamics and Chemistry RNA Structure and Dynamics > Influence of RNA Structure in Biological Systems.

Exploration and Characterization of Thermophilic Bacteria from the Kerinci-Jambi Geothermal Source for Alcohol Production

Exploration and Characterization of Thermophilic Bacteria from the Kerinci-Jambi Geothermal Source for Alcohol Production

Bioleaching of Mine Tailings by Mesophilic: Acidithiobacillus spp., Leptospirillum ferrooxidans, and Thermophilic: Sulfobacillus thermosulfidooxidans Cultures with the Addition of Ag+ Additive

This research focused on applying and comparing the performance of microorganisms with different temperature preferences, assessing the overall percentage recovery of elements (copper, zinc, arsenic, and iron) from mine tailings in the Staré Ransko region (Czech Republic). The study examined the solubilisation process using a mesophilic mixed bacterial culture, including Acidithiobacillus ferrooxidans (AF), Acidithiobacillus thiooxidans (AT), Leptospirillum ferrooxidans (LF), and the thermophilic species Sulfobacillus thermosulfidooxidans (ST). Under biotic conditions, constant process parameters were maintained, including a particle size of 71–100 µm, a pH value of 1.8, agitation at 150 rpm, and a pulp density of 10% (w/v). The only exception was the temperature, which varied for optimal multiplication of cultures (30 °C/50 °C). Additionally, the research examined the impact of AgNO3 additive at a concentration of Ag+ ions of 5 mg·L−1. The research focused on the solubilisation of Cu, Zn, Fe, and As and the results demonstrated that the application of microorganisms ST, combined with the action of Ag+ ions, enhanced the kinetics of the extraction process, leading to the highest final recovery of all elements (Cu 91.93%, Zn 85.67%, As 69.16%, and Fe 71.72%) under the specified conditions. The study observed that the most significant increase in solubilisation can be attributed to the additive cation in the case of copper (AF, AT, LF/Ag+ by 40.33%; ST/Ag+ by 44.39%) and arsenic (AF, AT, LF/Ag+ by 23.79%; ST/Ag+ by 26.08%). Notably, the intensification of leaching using thermophilic bacteria at a constant suspension temperature of 50 °C was primarily determined for Zn (ST by 18.36%, ST/Ag+ by 14.24%). After 24 days of extraction, the emergence of secondary minerals, namely CaSO4·2H2O and KFe3(SO4)2(OH)6, was identified. The study highlighted a significant increase in the extraction mechanism kinetics due to the influence of microorganisms, contrasting with the low solubilities observed under abiotic conditions (Cu 9.00%, Zn 14.17%, As 4.28%, Fe 6.23%).

Isolation of Thermophilic Bacteria from Extreme Environments in Northern Chile.

The northern region of Chile boasts unique geographical features that support the emergence of geothermal effluents, salt lagoons, and coastal creeks. These extreme climate conditions create polyextreme habitats for microorganisms, particularly adapted to survive these harsh environments. These extremophilic microorganisms hold immense potential as a source of hydrolytic enzymes, among other biotechnological applications. In this study, we isolated 15 strains of aerobic thermophilic bacteria (45-70 °C) from sediment samples collected at five different ecological sites, including hot springs, geothermal fields, and lagoons in the Atacama Desert and Andes high planes. Analyses of the 16S rRNA gene sequences of the isolates showed a close genetic similarity (98-100%) with microorganisms of the genera Parageobacillus, Geobacillus, Anoxybacillus, and Aeribacillus. Notably, these thermophiles exhibited significant hydrolytic enzyme activity, particularly amylases, lipases, and proteases. These findings underscore the potential of using these thermophilic bacterial strains as an invaluable source of thermozymes with wide-ranging applications in diverse industries, such as detergent formulations, pharmaceutical processing, and food technology. This research highlights the ecological significance of these extreme environments in the Atacama Desert and Andes high plains, which serve as vital ecological niches housing extremophilic bacteria as a genetic source of relevant thermozymes, promising great potential for innovation in the biotechnology industry.

When Synthesis Gets It Wrong: Unexpected Epimerization Using PyBOP in the Synthesis of the Cyclic Peptide Thermoactinoamide A.

Chemical synthesis is commonly seen as the final proof of the structure of complex natural products, but even a seemingly easy and well-established synthetic procedure may lead to an unexpected result. This is what happened with the synthesis of thermoactinoamide A (1a), an antimicrobial and antitumor nonribosomal cyclic hexapeptide produced by the thermophilic bacterium Thermoactinomyces vulgaris. The synthetic thermoactinoamide A outsourced to a company and the one described in a synthetic paper showed spectroscopic data identical to each other but different from those of the natural product. After a detailed spectroscopic, degradative, and synthetic study, the synthetic compound was shown to be an epimer (1b) of the intended target compound, originating during the cyclization reaction by extensive epimerization at the activated C-terminal amino acid. This allowed confirmation of the structure of the natural product.

Polycladomyces zharkentensis sp. nov., a novel thermophilic cellulose- and starch-degrading member of the Bacillota from a geothermal aquifer in Kazakhstan.

A thermophilic, aerobic and heterotrophic filamentous bacterium, designated strain ZKZ2T, was isolated from a pipeline producing hydrothermal water originating from a >2.3 km deep subsurface geothermal source in Zharkent, Almaty region, Kazakhstan. The isolate was Gram-stain-positive, non-motile, heat-resistant and capable of producing a variety of extracellular hydrolases. Growth occurred at temperatures between 55 and 75 °C, with an optimum around 70 °C, and at pH values between 5.5 and 9.0, with an optimum at pH 7.0-7.5 with the formation of aerial mycelia; endospores were produced along the aerial mycelium. The isolate was able to utilize the following substrates for growth: glycerol, l-arabinose, ribose, d-xylose, d-glucose, d-fructose, d-mannose, rhamnose, d-mannitol, methyl-d-glucopyranoside, aesculin, salicin, cellobiose, maltose, melibiose, sucrose, trehalose, melezitose, raffinose, starch, turanose and 5-keto-gluconate. Furthermore, it was able to hydrolyse carboxymethylcellulose, starch, skimmed milk, Tween 60 and Tween 80. The major cellular fatty acids were iso-C15 : 0, iso-C17 : 0, iso-C16 : 0 and C16 : 0. Our 16S rRNA gene sequence analysis placed ZKZ2T within the genus Polycladomyces, family Thermoactinomycetaceae, with the highest similarity to the type species Polycladomyces abyssicola JIR-001T (99.18 % sequence identity). Our draft genome sequence analysis revealed a genome size of 3.3 Mbp with a G+C value of 52.5 mol%. The orthologous average nucleotide identity value as compared to that of its closest relative, P. abyssicola JIR-001T, was 90.23 %, with an in silico DNA-DNA hybridization value of 40.7 %, indicating that ZKZ2T represents a separate genome species. Based on the phenotypic and genome sequence differences from the other two Polycladomyces species, we propose that strain ZKZ2T represents a novel species, for which we propose the name Polycladomyces zharkentensis sp. nov. The type strain is ZKZ2T (=CECT 30708T=KCTC 43421T).

Genome sequence of thermophilic cellulolytic actinobacterium Thermobifida fusca strain UPMC 901 producing thermostable cellulases.

Thermophilic cellulolytic bacteria producing thermostable cellulase have multiple potential industrial and biotechnological applications. In this study, we sequenced the genome of Thermobifida fusca UPMC 901 by using nanopore sequencing technology with 100 % genome coverage. The represented genome is 3.76 Mb with a G + C content of 67.4 %. The genome is composed of 1 scaffold with 3207 protein-coding genes. The results indicated that the genome of the Thermobifida fusca gene encodes at least 112 genes for carbohydrate-active enzymes. Of these genes, 40 glycoside hydrolases (GHs), 36 glycosyl transferases (GTs), 22 carbohydrate-binding modules (CBMs), 9 carbohydrate esterases (CEs), 3 polysaccharide lyases (PL) and 2 auxiliary activity (AAs) which refer to as lytic polysaccharide monooxygenase (LPMO), gene encoding the copper-dependent enzymes that catalyse oxidative cleavage of recalcitrant crystalline cellulose, were identified. In addition, two new functional genes, amylo-α-1,6 glucosidase and endo-α-1,4-mannosidase were also detected in the T. fusca genome. This study represented the ninth genome of the T. fusca and the cellulase genes identified will facilitate the exploration of potential sources for industrial and agricultural applications.

Using the sulfide-oxidizing bacterium Geobacillus thermodenitrificans to restrict H2S release during chicken manure composting

Hydrogen sulfide (H2S) is a toxic gas massively released during chicken manure composting. Diminishing its release requires efficient and low cost methods. In recent years, heterotrophic bacteria capable of rapid H2S oxidation have been discovered but their applications in environmental improvement are rarely reported. Herein, we investigated H2S oxidation activity of a heterotrophic thermophilic bacterium Geobacillus thermodenitrificans DSM465, which contains a H2S oxidation pathway composed by sulfide:quinone oxidoreductase (SQR) and persulfide dioxygenase (PDO). This strain rapidly oxidized H2S to sulfane sulfur and thiosulfate. The oxidation rate reached 5.73 μmol min−1·g−1 of cell dry weight. We used G. thermodenitrificans DSM465 to restrict H2S release during chicken manure composting. The H2S emission during composting process reduced by 27.5% and sulfate content in the final compost increased by 34.4%. In addition, this strain prolonged the high temperature phase by 7 days. Thus, using G. thermodenitrificans DSM465 to control H2S release was an efficient and economic method. This study provided a new strategy for making waste composting environmental friendly and shed light on perspective applications of heterotrophic H2S oxidation bacteria in environmental improvements.

The energy-converting hydrogenase Ech2 is important for the growth of the thermophilic acetogen Thermoanaerobacter kivui on ferredoxin-dependent substrates.

Acetogens thrive by converting H2+CO2 to acetate. Under environmental conditions, this allows for only very little energy to be conserved (∆G'<-20 kJ mol-1). CO2 serves as a terminal electron acceptor in the ancient Wood-Ljungdahl pathway (WLP). Since the WLP is ATP neutral, energy conservation during growth on H2 + CO2 is dependent on the redox metabolism. Two types of acetogens can be distinguished, Rnf- and Ech-type. The function of both membrane-bound enzyme complexes is twofold-energy conversion and redox balancing. Ech couples the Fd-dependent reduction of protons to H2 to the formation of a proton gradient in the thermophilic bacterium Thermoanaerobacter kivui. This bacterium may be utilized in gas fermentation at high temperatures, due to very high conversion rates and the availability of genetic tools. The physiological function of an Ech hydrogenase in T. kivui was studied to contribute an understanding of its energy and redox metabolism, a prerequisite for future industrial applications.

Produksi Enzim Spesifik Xilanase pada Variasi Suhu Konsorsium Trikultur Bakteri Termofilik

Xylanase is an extracellular enzyme that is capable of hydrolyzing xylan (hemicellulose) into xylose and xylo-oligosaccharides. Xylanase can be produced by microbes through a fermentation process, the microbes used are thermophilic bacteria. Thermophilic bacteria are microorganisms that are capable of producing thermostable enzymes through fermentation. The bacteria used are MSS11, MS 16 and MS 18. These thermophilic bacteria can live at temperatures above 45°C-80°C. To determine specific enzyme activity, enzyme and protein activity values were tested using a 540 nm (enzyme) and 750 nm (protein) septovotometer. The method used in this research is descriptive in triplicate. The research was carried out at the Microbiology Laboratory, Faculty of Mathematics and Natural Sciences, Padang State University on December 23 2023. Beechwood medium which had been mixed with MSS 11, MS 16, and MS 18 bacteria was then incubated at temperatures of 55℃, 65℃ and 75℃ so that specific enzyme activity was obtained. Of these three temperatures, the optimum specific enzyme activity of xylanase is 65℃, where the specific enzyme value obtained at 65℃ is 3.31 U/mg. Meanwhile, at low temperatures, namely at 55℃, the enzymatic reaction is slow, namely it has an enzyme activity value of 1.02 U/mg. Finally, at a temperature of 75, there was a decrease in specific enzyme activity, where the specific enzyme activity value was 3.12 U/mg.

Stabilization of biomass in manure effluent using the thermophilic-aerobic process

Aerobic-thermophilic stabilization of biomass is crucial in improving the processing processes of manure. This method makes it possible to achieve effective and stable processing of organic materials contained in manure using aerobic microorganisms and elevated temperatures. The aerobic-thermophilic process includes the action of oxygen and high temperature, which contribute to the active decomposition of organic substances in manure runoff. The increased temperature helps to create optimal conditions for the action of thermophilic bacteria, which accelerates the process of biological decomposition. Studies show that aerobic-thermophilic treatment of manure effluents helps to reduce the concentration of pathogenic microorganisms and ammonia. This contributes to obtaining cleaner and environmentally safe manure suitable for use in agriculture. Advantages of aerobic-thermophilic stabilization: Faster recycling process compared to traditional methods. Reduction of odors and risk of pathogen contamination. High quality and efficiency of produced manure. Aerobic-thermophilic stabilization of biomass in manure processing is a promising direction in improving modern waste treatment technologies. Its implementation contributes to improving the quality and safety of manure and the effective use of these resources in agriculture. Research and development of this method are essential for achieving sustainable and ecologically clean agricultural production. It has been established that liquid manure of enterprises with intensive milk and pork production technologies with hydraulic manure removal systems with a humidity of 92–96 % should be subjected to aerobic fermentation in an auto thermal mode, as a result of which products are obtained that are safe from a sanitary and hygienic point of view. In addition, the loss of nutrients is reduced. which determines the quality of organic fertilizers. Established that liquid manure businesses for technology-intensive milk production and pig manure removal by hydraulic systems with humidity 92–96 % of it are advisable to expose aerobic biofermentatsiyi in autothermal mode, which allows you to get the products in a safe sanitary and hygienic attitude and reduce the loss of nutrients that determine the quality of organic fertilizers.

Exploring the Hot Springs of Golan: A Source of Thermophilic Bacteria and Enzymes with Industrial Promise.

In recent years, there has been a surge in research on extremophiles due to their remarkable ability to survive in harsh environments. Extremophile thermophilic bacteria provide thermostable enzymes for biotechnology and industry. Thermophilic bacteria live in extreme environments like hot springs at 45-80°C. This study screens and isolates thermophilic bacteria and thermozymes from the Golan hot springs in Karakocan, Elazig, Turkey. The study also characterizes thermophilic bacteria and their thermozymes to understand their features and applications better. Golan hot spring water samples at 50°C yielded 12 isolates. GKE 02, 07, 08, and 10 produce amylase, GKE 04, 08, and 11 cellulase, and GKE 06 xylanase. One isolate (GKE 08) displayed both amylolytic and cellulolytic activity on agar plates. GKE 02 had the highest plate assay amylolytic index (2.3) and amylase activity (67.87 U/ml). Plate assay indicates GKE 08 has 1.5 amylolytic index, 1.1 cellulolytic index, 38.57 U/ml amylase, and 6.81 U/ml cellulase. GKE 04 had the greatest cellulolytic index (1.7) and cellulase activity (27.46). GKE 06, the only xylanase producer, has 19.67 U/ml activity and 1.4 plate assay index. The investigation also included determining the optimal pH and temperature conditions for each enzyme. 16S rDNA gene sequencing revealed seven thermozyme-producing bacteria Bacillus, Geobacillus, and Thermomonas. Thermomonas hydrothermalis genome annotation showed glycosyl hydrolase genes for amylolytic and cellulolytic activity. The findings of this study on thermophilic bacteria and thermostable enzyme synthesis in the Golan hot springs are promising, particularly for T. hydrothermalis, which has limited research on its potential as a thermozyme producer.

The Effects of Silver Nanoparticles (AgNPs) on Thermophilic Bacteria: Antibacterial, Morphological, Physiological and Biochemical Investigations.

Since thermophilic microorganisms are valuable sources of thermostable enzymes, it is essential to recognize the potential toxicity of silver nanoparticles used in diverse industrial sectors. Thermophilic bacteria Geobacillus vulcani 2Cx, Bacillus licheniformis 3CA, Paenibacillus macerans 3CA1, Anoxybacillus ayderensis FMB1, and Bacillus paralicheniformis FMB2-1 were selected, and their MIC and MBC values were assessed by treatment with AgNPs in a range of 62.5-1500 μg mL-1. The growth inhibition curves showed that the G. vulcani 2Cx, and B. paralicheniformis FMB2-1 strains were more sensitive to AgNPs, demonstrating a reduction in population by 71.1% and 31.7% at 62.5 μg mL-1 and by 82.9% and 72.8% at 250 μg mL-1, respectively. TEM and FT-IR analysis revealed that AgNPs caused structural damage, cytoplasmic leakage, and disruption of cellular integrity. Furthermore, cell viability showed a significant decrease alongside an increase in superoxide radical (SOR; O2-) production. β-galactosidase biosynthesis decreased to 28.8% level at 500 μg mL-1 AgNPs for G. vulcani 2Cx, 32.2% at 250 μg mL-1 for A. ayderensis FMB1, and 38.8% only at 62.5 μg mL-1, but it was completely inhibited at 500 μg mL-1 for B. licheniformis 3CA. Moreover, B. paralicheniformis FMB2-1 showed a significant decrease to 11.2% at 125 μg mL-1. This study is the first to reveal the toxic effects of AgNPs on thermophilic bacteria.

Screening of conditions for the acetic acid production from H2 and CO2 by Thermoanaerobacter kivui in a pressurized stirred tank bioreactor

Growing concerns regarding climate change have heightened interest in utilizing carbon dioxide as a valuable carbon source for chemical production. The acetogen Thermoanaerobacter kivui is a thermophilic bacterium that converts CO2 and H2 streams into acetic acid. Nevertheless, the limited solubility of the gaseous substrates in the liquid medium has often led to low productivity by the biocatalyst employed. This study aims to enhance the cell-specific acetic acid productivity of T. kivui by combining pressure, the composition of the inlet gas mix, and in-flow gas rate. Firstly, the combined effect of pressure and gas composition was assessed through a Design of Experiments approach. Tests were performed in a pressurized bioreactor and indicated that acetic acid cell-specific productivity was achieved at 10 bar, providing a 3:1 H2:CO2 blend. Subsequently, supplying this blend at high pressure into the vessel of the reactor, an in-flow gas rate screening was performed to identify completely the parameters that allowed the maximum acetic acid productivity. The optimal flow rate was 60 mL.min−1, and the acetic acid cell-specific productivity reached 2.90 g.g−1.h−1. Additionally to the experiments in the bioreactor, tests in serum bottles were performed to investigate the influence of the osmotic condition due to different salts and acetic acid inhibition on T. kivui. Results indicated that salts and acetic acid concentration impaired bacteria growth and affected the production of further acetic acid. Nevertheless, a metabolic shift toward the production of formic acid was observed specifically by adding the HCO3– ion.

Genomics of the Thermophilic Bacterium Thermosulfidibacter takaii Reveals Novel Lineage of Deep-Branching Bacterial Phylum.

The online version contains supplementary material available at 10.1007/s12088-024-01214-9.

Catch-and-Release: The Assembly, Immobilization, and Recycling of Redox-Reversible Artificial Metalloenzymes.

Technologies to improve the applicability of artificial metalloenzymes (ArMs) are gaining considerable interest; one such approach is the immobilization of these biohybrid catalysts on support materials to enhance stability and enable their retention, recovery, and reuse. Here, we describe the immobilization of polyhistidine-tagged ArMs that allow the redox-controlled replacement of catalytic cofactors that have lost activity, e.g., due to poisoning or decomposition, on immobilized metal affinity chromatography resins. By using periplasmic siderophore-binding protein scaffolds that originate from thermophilic bacteria (GstCeuE and PthCeuE) in combination with a siderophore-linked imine reduction catalyst, reaction rates were achieved that are about 3.5 times faster than those previously obtained with CjCeuE, the analogous protein of Campylobacter jejuni. Upon immobilization, the GstCeuE-derived ArM showed a decrease in turnover frequency in the reduction of dehydrosalsolidine by 3.4-fold, while retaining enantioselectivity (36%) and showing improved stability that allowed repeat recovery and recycling cycles. Catalytic activity was preserved over the initial four cycles. In subsequent cycles, a gradual reduction of activity was evident. Once the initial activity decreased to around 40% of the initial activity (23rd recycling cycle), the redox-triggered artificial cofactor release permitted the subsequent recharging of the immobilized protein scaffold with fresh, active cofactor, thereby restoring the initial catalytic activity of the immobilized ArM and allowing its reuse for several more cycles. Furthermore, the ArM could be assembled directly from protein present in crude cell extracts, avoiding time-consuming and costly protein purification steps. Overall, this study demonstrates that the immobilization of redox-reversible ArMs facilitates their "catch-and-release" assembly and disassembly and the recycling of their components, improving their potential commercial viability and environmental footprint.

Mesophiles vs. Thermophiles: Untangling the Hot Mess of Intrinsically Disordered Proteins and Growth Temperature of Bacteria.

The dynamic structures and varying functions of intrinsically disordered proteins (IDPs) have made them fascinating subjects in molecular biology. Investigating IDP abundance in different bacterial species is crucial for understanding adaptive strategies in diverse environments. Notably, thermophilic bacteria have lower IDP abundance than mesophiles, and a negative correlation with optimal growth temperature (OGT) has been observed. However, the factors driving these trends are yet to be fully understood. We examined the types of IDPs present in both mesophiles and thermophiles alongside those unique to just mesophiles. The shared group of IDPs exhibits similar disorder levels in the two groups of species, suggesting that certain IDPs unique to mesophiles may contribute to the observed decrease in IDP abundance as OGT increases. Subsequently, we used quasi-independent contrasts to explore the relationship between OGT and IDP abundance evolution. Interestingly, we found no significant relationship between OGT and IDP abundance contrasts, suggesting that the evolution of lower IDP abundance in thermophiles may not be solely linked to OGT. This study provides a foundation for future research into the intricate relationship between IDP evolution and environmental adaptation. Our findings support further research on the adaptive significance of intrinsic disorder in bacterial species.

Selective enhanced cytotoxicity of amino acid deprivation for cancer therapy using thermozyme functionalized nanocatalyst

BackgroundEnzyme therapy based on differential metabolism of cancer cells has demonstrated promising potential as a treatment strategy. Nevertheless, the therapeutic benefit of reported enzyme drugs is compromised by their uncontrollable activity and weak stability. Additionally, thermozymes with high thermal-stability suffer from low catalytic activity at body temperature, preventing them from functioning independently.ResultsHerein, we have developed a novel thermo-enzymatic regulation strategy for near-infrared (NIR)-triggered precise-catalyzed photothermal treatment of breast cancer. Our strategy enables efficient loading and delivery of thermozymes (newly screened therapeutic enzymes from thermophilic bacteria) via hyaluronic acid (HA)-coupled gold nanorods (GNRs). These nanocatalysts exhibit enhanced cellular endocytosis and rapid enzyme activity enhancement, while also providing biosafety with minimized toxic effects on untargeted sites due to temperature-isolated thermozyme activity. Locally-focused NIR lasers ensure effective activation of thermozymes to promote on-demand amino acid deprivation and photothermal therapy (PTT) of superficial tumors, triggering apoptosis, G1 phase cell cycle arrest, inhibiting migration and invasion, and potentiating photothermal sensitivity of malignancies.ConclusionsThis work establishes a precise, remotely controlled, non-invasive, efficient, and biosafe nanoplatform for accurate enzyme therapy, providing a rationale for promising personalized therapeutic strategies and offering new prospects for high-precision development of enzyme drugs.Graphical