Ultrahigh Heat/Fire-Resistant, Mechanically Robust, and Closed-Loop Chemical Recyclable Polycarbonate Enabled by Facile Bond Dissociation Energy Modulation.

Plastics serve as an essential foundation in contemporary society. Nevertheless, meeting the rigorous performance demands in advanced applications and addressing their end-of-life disposal are two critical challenges that persist. Here, an innovative and facile method is introduced for the design and scalable production of polycarbonate, a key engineering plastic, simultaneously achieving high performance and closed-loop chemical recyclability. The bisphenol framework of polycarbonate is strategically adjusted from the low-bond-dissociation-energy bisphenol A to high-bond-dissociation-energy 4,4'-dihydroxydiphenyl, in combination with the incorporation of polysiloxane segments. As expected, the enhanced bond dissociation energy endows the polycarbonate with an extremely high glow-wire flammability index surpassing 1025 °C, a 0.8 mm UL-94 V-0 rating, a high LOI value of 39.2%, and more than 50% reduction of heat and smoke release. Furthermore, the π-π stacking interactions within biphenyl structures resulted in a significant enhancement of mechanical strength by as more as 37.7%, and also played a positive role in achieving a lower dielectric constant. Significantly, the copolymer exhibited outstanding closed-loop chemical recyclability, allowing for facile depolymerization into bisphenol monomers and the repolymerized copolymer retains its high heat and fire resistance. This work provides a novel insight in the design of high-performance and closed-loop chemical recyclable polymeric materials.

Progress in using cross-linking technologies to increase the thermal stability of colloidal delivery systems.

In recent years, crosslinking technology has been found and widely used in food, textile, pharmaceutical, bioengineering and other fields. Crosslinking is a reaction in which two or more molecules bond to each other to form a stable three-dimensional network structure to improve the strength, heat resistance and other properties of substances. The researchers found that the cross-linking technology has a significant effect on improving the thermal stability of the colloidal delivery system. In this paper, crosslinking techniques that can be used to improve the thermal stability of colloidal delivery systems are reviewed, including enzyme-, ion-, chemical-, and combined cross-linking. Initially, the underlying mechanisms of these crosslinking technologies is reviewed. Then, the impacts of crosslinking on the heat-stability of colloidal delivery systems are discussed. Finally, the application of crosslinked delivery systems in improving the thermal stability of probiotics, polyphenols, pigments, and nutrients in foods and food packaging materials is introduced. The ability of proteins and polysaccharides to form heat-stable colloidal delivery systems can be improved by crosslinking. Nevertheless, more research is required to establish the impact of different crosslinking on the thermal stability of a broader range of different delivery systems, as well as to ensure their safety and efficacy.

Thermally Stable and Transparent Polyimide Derived from Side-Group-Regulated Spirobifluorene Unit for Substrate Application.

Advancements in flexible electronic technology, especially the progress in foldable displays and under-display cameras (UDC), have created an urgent demand for high-performance colorless polyimide (CPI). However, current CPIs lack sufficient heat resistance for substrate applications. In this work, four kinds of rigid spirobifluorene diamines are designed, and the corresponding polyimides are prepared by their condensation with 5,5'-(perfluoropropane-2,2-diyl) bis(isobenzofuran-1,3-dione) (6FDA) or 9,9-bis(3,4-dicarboxyphenyl) fluorene dianhydride (BPAF). The rigid and conjugated spirobifluorene units endow the polyimides with higher glass transition temperature (Tg) ranging from 356 to 468 °C. Their optical properties are regulated by small side groups and spirobifluorene structure with a periodically twisted molecular conformation. Consequently, a series of CPIs with an average transmittance ranging from 75% to 88% and a yellowness index (YI) as low as 2.48 are obtained. Among these, 27SPFTFA-BPAF presents excellent comprehensive performance, with a Tg of 422 °C, a 5 wt.% loss temperature (Td5) of 562 °C, a YI of 3.53, and a tensile strength (δmax) of 140 MPa, respectively. The mechanism underlying the structure-property relationship is investigated by experimental comparison and theoretical calculation, and the proposed method provides a pathway for designing highly rigid conjugated CPIs with excellent thermal stability and transparency for photoelectric engineering.

Design of Co-Cured Multi-Component Thermosets with Enhanced Heat Resistance, Toughness, and Processability via a Machine Learning Approach.

Designing heat-resistant thermosets with excellent comprehensive performance has been a long-standing challenge. Co-curing of various high-performance thermosets is an effective strategy, however, the traditional trial-and-error experiments have long research cycles for discovering new materials. Herein, a two-step machine learning (ML) assisted approach is proposed to design heat-resistant co-cured resins composed of polyimide (PI) and silicon-containing arylacetylene (PSA), that is, poly(silicon-alkyne imide) (PSI). First, two ML prediction models are established to evaluate the processability of PIs and their compatibility with PSA. Then, another two ML models are developed to predict the thermal decomposition temperature and flexural strength of the co-cured PSI resins. The optimal molecular structures and compositions of PSI resins are high-throughput screened. The screened PSI resins are experimentally verified to exhibit enhanced heat resistance, toughness, and processability. The research framework established in this work can be generalized to the rational design of other advanced multi-component polymeric materials.

Differential impacts of nonsteroidal anti-inflammatory drugs on lifespan and healthspan in aged Caenorhabditis elegans.

Aging and age-related diseases are intricately associated with oxidative stress and inflammation. Nonsteroidal anti-inflammatory drugs (NSAIDs) have shown their promise in mitigating age-related conditions and potentially extending lifespan in various model organisms. However, the efficacy of NSAIDs in older individuals may be influenced by age-related changes in drug metabolism and tolerance, which could result in age-dependent toxicities. This study aimed to evaluate the potential risks of toxicities associated with commonly used NSAIDs (aspirin, ibuprofen, acetaminophen, and indomethacin) on lifespan, healthspan, and oxidative stress levels in both young and old Caenorhabditis elegans. The results revealed that aspirin and ibuprofen were able to extend lifespan in both young and old worms by suppressing ROS generation and enhancing the expression of antioxidant SOD genes. In contrast, acetaminophen and indomeacin accelerated aging process in old worms, leading to oxidative stress damage and reduced resistance to heat stress through the pmk-1/skn-1 pathway. Notably, the harmful effects of acetaminophen and indomeacin were mitigated when pmk-1 was knocked out in the pmk-1(km25) strain. These results underscore the potential lack of benefit from acetaminophen and indomeacin in elderly individuals due to their increased susceptibility to toxicity. Further research is essential to elucidate the underlying mechanisms driving these age-dependent responses and to evaluate the potential risks associated with NSAID use in the elderly population.

Temperature adaptation patterns in Chinese cattle revealed by TRPM2 gene mutation analysis.

Cattle are sensitive to temperature fluctuations but adapt well to inclement weather conditions. When environmental temperatures exceed specific thresholds, heat stress becomes a critical concern for cattle. The TRPM2 gene, which resides on cattle chromosome 1 encodes a TRP channel protein, holding a unique capacity to sense temperature changes and facilitate rapid response to avoid heat stress. Here, we utilized the Bovine Genome Variation Database (BGVD) (http://animal.omics.pro/code/index.php/BosVar), and identified a missense mutation site, c.805A > G: p. Met269Val (rs527146862), within the TRPM2 gene. To elucidate the functional assessment of this mutation in temperature adaptation attributes of Chinese cattle, we genotyped 407 samples from 20 distinct breeds representing diverse climatic zones across China. The association analysis incorporates three temperature parameters and revealed compelling insights in terms of allele frequency. Interestingly, the prevalence of the wild-type allele A was notably higher among northern cattle breeds and this trend diminished gradually as observed in southern cattle populations. Conversely, the mutant-type allele G demonstrated a contrasting trend. Moreover, southern cattle exhibited markedly higher frequencies of GG and GA genotypes (P < 0.01). The presence of heterozygous and homozygous mutations appears to confer an enhanced capacity for adaptation to elevated temperatures. These results provide unequivocal correlation evidence between TRPM2 genotypes (AA, GA, GG) and environmental temperature parameters and comprehend the genetic mechanisms governing temperature adaptation in cattle. This provides valuable insights for strategic breed selection across diverse climatic regions, thereby aiding livestock production amid evolving climate challenges.

The TRPM2 gene encodes TRP channel protein that helps animals in combating heat stress. Twenty Chinese local cattle breeds were genotyped, and association analysis was performed. This investigation encompasses the distribution pattern of the missense mutation locus rs527146862 of the TRPM2 gene in southern, northern, and central cattle populations. The results demonstrated a significant relationship between rs527146862 locus and temperature adaptation attributes in Chinese cattle.

Metal-organic framework adhesives with exceptionally high heat resistance.

We synthesized high-heat-resistant adhesives based on metal - organic frameworks owing to their high decomposition temperature and the absence of a glass transition. Heat-resistance tests were performed on adhesive joints consisting of zeolitic imidazolate framework (ZIF)-67-based adhesives and a copper substrate. The as-synthesized ZIF-67-based adhesive exhibited heat resistances at 600 and 700°C in air and nitrogen atmospheres, respectively, comparable to those of conventional high-heat-resistant polymer-based adhesives. The degradation mechanism of the ZIF-67 adhesives was investigated, and their high heat resistance was attributed to the stable existence of the ZIF-67 qtz phase in the adhesive layer at high temperatures without the formation of voids. Thus, adhesives based on ZIF-67 and other metal - organic frameworks can be applied in high-temperature industrial systems.

By focusing on its high thermal stability and absence of glass transition, the ZIF-67 gel was found to have high potential that is comparable to existing heat-resistant adhesives.

Stress-Tolerant Dormant Bacterial Forms: Biological and Ultrastructural Properties of Moraxella catarrhalis and Kocuria rhizophila.

Dormant forms of causative agents of healthcare-acquired infections Moraxella catarrhalis and Kocuria rhizophila have been obtained. Dormant forms cells retained viability during long-term storage (≈107 CFU/ml after 2 months) under provocative conditions (lack of nutrient sources; temperature 20°C, oxygen access) were characterized by heat resistance, and acquired special ultrastructural organization typical of dormant forms (compacted nucleoid, thickened cell wall). They were also capable of forming alternative phenotypes (dominant and small colony variants) in a new cycle of germination in a fresh medium. These results demonstrate that the dormant forms can be responsible both for survival in the environment and persistence in the host organism.

Development of heat-shock resistance in Legionella pneumophila modeled by experimental evolution.

Because it can grow in buildings with complex hot water distribution systems (HWDS), healthcare facilities recognize the waterborne bacterium Legionella pneumophila as a major nosocomial infection threat and often try to clear the systems with a pasteurization process known as superheat-and-flush. After this treatment, many facilities find that the contaminating populations slowly recover, suggesting the possibility of in situ evolution favoring increased survival in high-temperature conditions. To mimic this process in a controlled environment, an adaptive laboratory evolution model was used to select a wild-type strain of L. pneumophila for survival to transient exposures to temperatures characteristic of routine hot water use or failed pasteurization processes in HWDS. Over their evolution, these populations became insensitive to exposure to 55°C and developed the ability to survive short exposures to 59°C heat shock. Heat-adapted lineages maintained a higher expression of heat-shock genes during low-temperature incubation in freshwater, suggesting a pre-adaptation to heat stress. Although there were distinct mutation profiles in each of the heat-adapted lineages, each acquired multiple mutations in the DnaJ/DnaK/ClpB disaggregase complex, as well as mutations in chaperone htpG and protease clpX. These mutations were specific to heat-shock survival and were not seen in control lineages included in the experimental model without exposure to heat shock. This study supports in situ observations of adaptation to heat stress and demonstrates the potential of L. pneumophila to develop resistance to control measures. IMPORTANCE As a bacterium that thrives in warm water ecosystems, Legionella pneumophila is a key factor motivating regulations on hot water systems. Two major measures to control Legionella are high circulating temperatures intended to curtail growth and the use of superheat-and-flush pasteurization processes to eliminate established populations. Facilities often suffer recolonization of their hot water systems; hospitals are particularly at risk due to the severe nosocomial pneumoniae caused by Legionella. To understand these long-term survivors, we have used an adaptive laboratory evolution model to replicate this process. We find major differences between the mutational profiles of heat-adapted and heat-naïve L. pneumophila populations including mutations in major heat-shock genes like chaperones and proteases. This model demonstrates that well-validated treatment protocols are needed to clear contaminated systems and-in an analog to antibiotic resistance-the importance of complete eradication of the resident population to prevent selection for more persistent bacteria.

Optimization of food-grade colloidal delivery systems for thermal processing applications: a review.

Colloidal delivery systems are widely used in the food industry to enhance the dispersibility, stability, efficacy, or bioavailability. However, when exposed to the high temperature, delivery systems are often prone to degradation, which limits its application in thermal processing. In this paper, the effects of thermal processing on the performance of traditional protein-based or starch-based delivery systems are firstly described, including the molecular structure changes of proteins, starches or lipids, and the degradation of embedded substances. These effects are unfavorable to the application of the delivery system in thermal processing. Then, strategies of improving the heat resistance of food grade colloid delivery system and their use in frying, baking and cooking food are mainly introduced. The heat resistance of the delivery system can be improved by a variety of strategies, including the development of new heat-resistant materials, the addition of heat-resistant coatings to the surface of delivery systems, the cross-linking of proteins or starches using cross-linking agents, the design of particle structures, the use of physical means such as ultrasound, or the optimization of the ingredient formula. These strategies will help to expand the application of heat-resistant delivery systems so that they can be used in real thermal processing.

Purification and enzymatic properties of a new thermostable endoglucanase from Aspergillus oryzae HML366.

Aspergillus oryzae HML366 is a newly screened cellulase-producing strain. The endoglucanase HML ED1 from A. oryzae HML366 was quickly purified by a two-step method that combines ammonium sulfate precipitation and strong anion exchange column. SDS-PAGE electrophoresis indicated that the molecular weight of the enzyme was 68 kDa. The optimum temperature of the purified endoglucanase was 60 ℃ and the enzyme activity was stable below 70 ℃. The optimum pH was 6.5, and the enzyme activity was stable at pH between 4.5 and 9.0. The analysis indicated that additional Na+, K+, Ca2+, and Zn2+ reduced the catalytic ability of enzyme to the substrate, but Mn2+ enhanced its catalytic ability to the substrate.The Km and Vmax of the purified endoglucanase were 8.75 mg/mL and 60.24 µmol/min·mg, respectively. In this study, we report for the first time that A. oryzae HML366 can produce a heat-resistant and wide pH tolerant endoglucanase HML ED1, which has potential industrial application value in bioethanol, paper, food, textile, detergent, and pharmaceutical industries.

UV-C and wet heat resistance of Bacillus thuringiensis biopesticide endospores compared to foodborne Bacillus cereus endospores.

Bacillus endospores (spores) are generally resistant to environmental and food processing-related stress including thermal and non-thermal processing in the food industry, such as pasteurization, and UV-C inactivation. Bacillus thuringiensis insecticidal crystals and spores as the active substances in commercial biopesticides can also be introduced to vegetable foods and their food processing environment due to pre-harvest treatment of edible crops. The resistance of B. thuringiensis biopesticide spores in comparison to the genetically closely related foodborne B. cereus against heat and UV-C treatment is investigated in this study. The results show that B. thuringiensis biopesticide spores with the commercial granulated product formulation are better protected and as such more resistant to both wet heat (D values at 90 °C: 50.1-79.5 min) and UV-C treatment (D values at 0.6 mW/cm2: 7.5-8.9 min) than the pure spore suspension. The enhanced UV-C resistance properties of B. thuringiensis-formulated spores also indicate that the B. thuringiensis spores in powder or granule formulation applied in the field might not be effectively inactivated by solar radiation (UV-A and UV-B) in a short period. Furthermore, the spores of one emetic B. cereus toxin-producing strain (LFMFP 254; a Belgian outbreak strain) were found more resistant to the wet heat at 90 °C (D90-value = 71.2 min) than other tested pure spore suspensions, although the spores of B. cereus 254 did not show different behavior against UV-C treatment. This result suggests that UV-C treatment can be applied as an effective inactivation method against B. cereus 254 spores.

Weak sex-specific evolution of locomotor activity of Sepsis punctum (Diptera: Sepsidae) thermal experimental evolution lines.

Elevated temperatures are expected to rise beyond what the physiology of many organisms can tolerate. Behavioural responses facilitating microhabitat shifts may mitigate some of this increased thermal selection on physiology, but behaviours are themselves mediated by physiology, and any behavioural response may trade-off against other fitness-related activities. We investigated whether experimental evolution in different thermal regimes (Cold: 15 °C; Hot: 31 °C; Intergenerational fluctuation 15/31 °C; Control: 23 °C) resulted in genetic differentiation of standard locomotor activity in the dung fly Sepsis punctum. We assessed individual locomotor performance, an integral part of most behavioral repertoires, across eight warm temperatures from 24 °C to 45 °C using an automated device. We found no evidence for generalist-specialist trade-offs (i.e. changes in the breadth of the performance curve) for this trait. Instead, at the warmest assay temperatures hot-selected flies showed somewhat higher maximal performance than all other, especially cold-selected flies, overall more so in males than females. Yet, the flies' temperature optimum was not higher than that of the cold-selected flies, as expected under the 'hotter-is-better' hypothesis. Maximal locomotor performance merely weakly increased with body size. These results suggest that thermal performance curves are unlikely to evolve as an entity according to theory, and that locomotor activity is a trait of limited use in revealing thermal adaptation.

Effects of High Temperature Stress on the Physiological and Biochemical Characteristics of Paeonia ostii.

In order to explore the effects of high temperature stress on the physiological characteristics of Paeonia ostii, the Paeonia ostii were subjected to 25 °C, 35 °C, 38 °C, and 40 °C for 7 days. Meanwhile, the physiological indicators of oxidative stress (hydrogen peroxide, H2O2; malondialdehyde, MDA; relative electrical conductivity, REC), antioxidant enzyme activity (superoxide dismutase, SOD; ascorbate peroxidase, APX; catalase, CAT; peroxidase, POD), photosynthetic pigment content (chlorophyll a, Chla; chlorophyll b, Chlb), photosynthetic characteristics (net photosynthetic rate, Pn; intercellular CO2 concentration, Ci; stomatal conductance, Gs; transpiration rate, Tr), and osmoregulatory substances content (soluble protein, SP; soluble sugar, SS) were determined. The results showed that, with the increase in temperature and stress time, the H2O2 content, MDA content, REC value, CAT activity, and APX activity increased, while Chla content, Chlb content, SS content, and SP content decreased. With the extension of stress time, the SOD activity, POD activity, and Tr value of each high temperature stress group first increased and then decreased; Ci first decreased, then increased, and then decreased; meanwhile, Pn and Gs showed an overall downward trend. PLS-DA (partial least squares discriminant analysis) was used to analyze the changes in physiological and biochemical indexes of peony leaves under 40 °C stress for different days. SOD was found to be the biggest factor affecting the changes in physiological and biochemical indexes of peony leaves treated with different days of stress.

Er-Based Luminescent Nanothermometer to Explore the Real-Time Temperature of Cells under External Stimuli.

Temperature as a typical parameter, which influences the status of living creatures, is essential to life activities and indicates the initial cellular activities. In recent years, the rapid development of nanotechnology provides a new tool for studying temperature variation at the micro- or nano-scales. In this study, an important phenomenon is observed at the cell level using luminescent probes to explore intracellular temperature changes, based on Yb-Er doping nanoparticles with special upconversion readout mode and intensity ratio signals (I525 and I545 ). Further optimization of this four-layer core-shell ratio nanothermometer endows it with remarkable characteristics: super photostability, sensitivity, and protection owing to the shell. Thus this kind of thermal probe has the property of anti-interference to the complex chemical environment, responding exclusively to temperature, when it is used in liquid and cells to reflect external temperature changes at the nanoscale. The intracellular temperature of living RAW and CAOV3 cells are observed to have a resistance mechanism to external stimuli and approach a more favorable temperature, especially for CAOV3 cells with good heat resistance, with the intracellular temperature 4.8 °C higher than incubated medium under 5 °C environment, and 4.4 °C lower than the medium under 60 °C environment.

Genome size of alpine plants does not predict temperature resistance.

MAIN CONCLUSION: Genome size of alpine plants is not related to their resistance against frost and heat. Genome size is a variable trait in angiosperms, and it was suggested that large genome size represents a constraint in stressful environments. We measured genome size and resistance to frost and heat in 89 species of plants from tropical and temperate alpine habitats. Genome size of the species, ranging from 0.49 pg to 25.8 pg across the entire dataset, was not related to either frost or heat resistance in either group of plants. Genome size does not predict resistance to extreme temperatures in alpine plants and is thus not likely to predict plant responses to climate changes.

Identification of the DcHsp20 gene family in carnation (Dianthus caryophyllus) and functional characterization of DcHsp17.8 in heat tolerance.

MAIN CONCLUSION: 33 heat shock protein 20 (Hsp20) genes were identified from the carnation genome whose expression were altered by abiotic stresses. DcHsp17.8 may function to improve the heat resistance of Arabidopsis. Heat shock proteins 20 (Hsp20s) mainly function as molecular chaperones that play crucial roles in relieving abiotic stresses such as heat stress. In this study, we identified and characterized 33 DcHsp20 genes from the carnation genome that were classified into 9 subfamilies. Gene structure analysis showed that 25 DcHsp20 genes contained 1 intron whilst the remaining 8 DcHsp20 genes did not contain introns. Motif analysis found that DcHsp20 proteins were relatively conserved. Cis-regulatory elements analysis of the Hsp20 promoters revealed a number of cis-regulatory elements that regulate growth and development, hormone and stress responses. Gene expression analysis revealed that DcHsp20 genes had multiple response patterns to heat stress. The largest range of induction occurred in DcHsp17.8 after 1 h of heat stress. Under cold stress, or treatment with saline or abscisic acid, the expression of most DcHsp20 genes was inhibited. To further understand the function of DcHsp20 genes in response to heat stress, we overexpressed DcHsp17.8 in Arabidopis and the plants showed improved heat tolerance, O2- and H2O2 activities and photosynthetic capacity with reduced relative electrolyte leakage and malondialdehyde content. Gene expression analysis revealed that DcHsp17.8 modulated the expression of genes involved in antioxidant enzyme synthesis. Our data provided a solid foundation for the further detailed study of DcHsp20 genes.

Genetic Characteristics of the Transmissible Locus of Stress Tolerance (tLST) and tLST Harboring Escherichia coli as Revealed by Large-Scale Genomic Analysis.

The transmissible locus of stress tolerance (tLST) confers resistance to multiple stresses in E. coli. Utilizing 18,959 E. coli genomes available in the NCBI database, we investigated the prevalence, phylogenetic distribution, and configuration patterns of tLST, and correlations between tLST, and virulence and antimicrobial resistance (AMR) genes in E. coli. Four tLST variants were found in 2.7% of E. coli, with the most prevalent (77.1%) variant being tLST1 followed by tLST2 (8.3%), tLST3b (8.3%) and tLST3a (6.3%). The majority (93%) of those tLST were in E. coli belonging to phylogroup A in which the prevalence was 10.4%. tLST was also found in phylogroup B1 (0.5%) and C (0.5%) but not found in B2 or D-G. An additional 1% of the 18,959 E. coli genomes harbored tLST fragments to various extent. Phylogenetic analysis revealed both intra- and interspecies transmission of both chromosomal and plasmid-borne tLST, with E. coli showing a preference of chromosomal over plasmid-borne tLST. The presence of tLST and virulence genes in E. coli was overall negatively correlated, but tLST was found in all genomes of a subgroup of enterotoxigenic E. coli (ST2332). Of note, no Shiga toxin-producing E. coli (n = 3,492) harbored tLST. The prevalence of tLST and AMR genes showed different temporal trends over the period 1985 to 2019. However, a substantial fraction of tLST positive E. coli harbor AMR genes, posing a threat to public health. In conclusion, this study improves our understanding of the genetic characteristics of tLST and E. coli harboring tLST. IMPORTANCE This study, through a large-scale genomic analysis, demonstrated that the genomic island tLST related to multiple stress resistance (such as extreme heat resistance and oxidative stress tolerance) in E. coli is differentially present in subgroups of E. coli and is strongly associated with certain phylogenetic background of the host strain. The study also shows the transmission mechanisms of tLST in E. coli and other bacterial species. The overall negative association of tLST, and virulence genes and antimicrobial (AMR) genes suggest the selective pressures for the acquisition and transmission of these traits likely differ. Even so, the high prevalence of tLST in the enterotoxigenic E. coli clone ST2332 and co-occurrence of tLST and AMR genes in E. coli are concerning. Thus, the findings better our understanding of tLST evolution and provide information for risk assessment of tLST harboring bacteria.

Rational design of a disulfide bridge increases the thermostability of microbial transglutaminase.

Microbial transglutaminase (MTG) has numerous industrial applications in the food and pharmaceutical sectors. Unfortunately, the thermostability of MTG is too low to tolerate the desired conditions used in many of these commercial processes. In a previous study, we used protein engineering to improve the thermostability of MTG. Specifically, we generated a T7C/E58C mutant of MTG from Streptomyces mobaraensis that displayed enhanced resistance to thermal inactivation. In this study, a rational structure-based approach was adopted to introduce a disulfide bridge to further increase the thermostability of MTG. In all, four new mutants, each containing a novel disulfide bond, were engineered. Of these four mutants, D3C/G283C showed the most promising thermostability with a significantly higher ∆T50 (defined as the temperature of incubation at which 50% of the initial activity remains) of + 9 °C by comparison to wild-type MTG. Indeed, D3C/G283C combined enhanced thermostability with a 2.1-fold increased half-life at 65 °C compared with the wild-type enzyme. By structure-based rational design, we were able to create an MTG variant which might be useful for expanding the scope of application in food. KEY POINTS: • Microbial transglutaminase (MTG) is an enzyme used in many food applications • The applicability of MTG to various industrial processes other than the food sector is being investigated • Improvement of thermostability was confirmed for the disulfide bridge mutant D3C/G283C.

Geobacillus stearothermophilus STCC4517 spore suspensions showed survival curves with shoulder phenomena independent of sporulation temperature and pH, whose duration was an exponential function of treatment temperature.

Deviations in linearity in survival curves are common in inactivation kinetics during heat treatment. These might lead one to underestimate how effective thermal treatment is. In previous research we reported a relationship between decimal reduction time values (DT) and shoulder lengths (Sl) of survival curves which was characteristic of each microorganism. However, the impact of other factors such as sporulation temperature and pH of the treatment media on shoulder length is still not known. The objective of this research was to evaluate the effect of sporulation temperature (45, 55 and 65 °C) and pH (4.0, 5.0, 6.0 and 7.0) treatment has on the profile of survival curves and on the relationship between Sl/DT of G. stearothermophilus STCC 4517. The results obtained demonstrated that all the spore suspensions, independently of sporulation temperature and pH, showed survival curves with shoulder phenomena, whose duration was an exponential function of treatment temperature. Although both parameters had a significant effect on heat resistance, the relationship between the shoulder length and DT values was constant at all pHs for spores produced at the optimum sporulation temperature.