Kinetic Constraints Research Articles

Molecular Mechanisms of Active Transport in Antiporters: Kinetic Constraints and Efficiency.

A vital role in supporting successful functioning of biological cells is played by membrane channels called antiporters. These channel proteins utilize the concentration gradient of one type of species to move another type of species in the opposite direction and against their concentration gradient. It is believed that antiporters operate via alternating conformational transitions that expose these proteins to different sides of the membrane, and that only thermodynamics controls the activation of these channels. Here we explicitly investigate a chemical-kinetic model of antiporters to argue that there are additional kinetic constraints that need to be satisfied for these channels to be operational. This implies that kinetics and not thermodynamics governs the functioning of antiporters. In addition, the efficiency of antiporters is analyzed and the most optimal operating conditions are discussed. Our theoretical analysis clarifies some important aspects of the molecular mechanisms of biological membrane transport.

Constraint-induced breaking and restoration of ergodicity in spin-1 PXP models

Eigenstate Thermalization Hypothesis(ETH) has played a pivotal role in understanding ergodicity and its breaking in isolated quantum many-body systems. Recent experiment on 51-atom Rydberg quantum simulator and subsequent theoretical analysis have shown that hardcore kinetic constraint can lead to weak ergodicity breaking. In this work, we demonstrate, using 1d spin-1 PXP chains, that miscellaneous type of ergodicity can be realized by adjusting the hardcore constraints between different components of nearest neighbor spins. This includes ETH violation due to emergent shattering of Hilbert space into exponentially many subsectors of various sizes, a novel form of non-integrability with an extensive number of local conserved quantities and strong ergodicity. We analyze these different forms of ergodicity and study their impact on the non-equilibrium dynamics of a Z2 initial state. We use forward scattering approximation (FSA) to understand the amount of Z2-oscillation present in these models. Our work shows that not only ergodicity breaking but an appropriate choice of constraints can lead to restoration of ergodicity as well.

Evolution of human respiratory virus epidemics.

Background: Pathogens are often assumed to evolve towards reduced virulence, but counterexamples abound. Faced with a new pathogen, such as SARS-CoV-2, it is crucial to be able to forecast the case fatality rate (CFR) and the overall disease burden. Considerable effort has been invested towards developing a mathematical framework for predicting virulence evolution. Although many approaches accurately recapitulate complex outcomes, most rely on an assumed trade-off between CFR and infection rate. It is often impractical to empirically validate this constraint for human pathogens. Methods: A compartment model with parameters tuning the degree to which symptomatic individuals are isolated and the duration of immunity is constructed and evaluated at both short timescales and at equilibrium. Results: We reveal kinetic constraints whereby variation of multiple parameters in concert leads to decreased CFR and increased pathogen fitness, whereas independent variation of the parameters decreases pathogen fitness. Smallpox, SARS-CoV-2, and influenza are analyzed as diverse representatives of human respiratory viruses. We show that highly virulent viruses, such as smallpox, are often constrained by the host behavior, whereas moderately virulent viruses, such as SARS-CoV-2, appear to be typically constrained by the relationship between the duration of immunity and CFR. Conclusions: Evolution of human respiratory epidemics appears to be often kinetically constrained and a reduction in CFR should not be assumed. These results agree with previous work demonstrating an increase in virulence for smallpox and further predict that SARS-CoV-2 is likely to continue presenting a substantial disease burden. Herd immunity against SARS-CoV-2 and viruses with similar life history traits might be unachievable without vaccination. However, partial isolation of symptomatic individuals can have a major effect on the epidemic dynamics not only by reducing the number of fatalities in the short term but also by changing the evolutionary trajectory of moderate CFR viruses towards reduced CFR.

Observing non-ergodicity due to kinetic constraints in tilted Fermi-Hubbard chains

The thermalization of isolated quantum many-body systems is deeply related to fundamental questions of quantum information theory. While integrable or many-body localized systems display non-ergodic behavior due to extensively many conserved quantities, recent theoretical studies have identified a rich variety of more exotic phenomena in between these two extreme limits. The tilted one-dimensional Fermi-Hubbard model, which is readily accessible in experiments with ultracold atoms, emerged as an intriguing playground to study non-ergodic behavior in a clean disorder-free system. While non-ergodic behavior was established theoretically in certain limiting cases, there is no complete understanding of the complex thermalization properties of this model. In this work, we experimentally study the relaxation of an initial charge-density wave and find a remarkably long-lived initial-state memory over a wide range of parameters. Our observations are well reproduced by numerical simulations of a clean system. Using analytical calculations we further provide a detailed microscopic understanding of this behavior, which can be attributed to emergent kinetic constraints.

Impurity effect on recrystallization and grain growth in severe plastically deformed copper

The impurity effect on commercial purity copper (4 N and 3 N) was investigated by comparing their annealing induced recrystallization and grain growth behavior after severe plastic deformation. Equal channel angular pressing (ECAP) with additional cryogenic rolling was applied to produce equiaxed ultra-fine grained (UFG) and nano-laminated (NL) coppers with high stored excess energy. The stored excess energy increased with decreasing purity from 4 N to 3 N, whereas the grain boundary velocity and recrystallized grain size decreased for both UFG and NL Cu. In particular, the NL structures were found to be more sensitive to impurities than equiaxed ultra-fine grained ones, and consequently the abnormal grain growth at the early stages of recrystallization in NL 4 N Cu was effectively suppressed in NL 3 N Cu. The analysis of the grain growth rate during recrystallization, using the microstructural path methodology (MPM), demonstrates that the correlation between the abnormal grain growth and the significantly enhanced grain boundary mobility cannot be solely explained by the high stored excess energy. Under all conditions, the impurity drag effect poses a kinetic constraint on the rate-controlling mechanism for grain boundary migration.

Evolution of human respiratory virus epidemics

Background: Pathogens are often assumed to evolve towards reduced virulence, but counterexamples abound. Faced with a new pathogen, such as SARS-CoV-2, it is crucial to be able to forecast the case fatality rate (CFR) and the overall disease burden. Considerable effort has been invested towards developing a mathematical framework for predicting virulence evolution. Although many approaches accurately recapitulate complex outcomes, most rely on an assumed trade-off between CFR and infection rate. It is often impractical to empirically validate this constraint for human pathogens.Methods: A compartment model with parameters tuning the degree to which symptomatic individuals are isolated and the duration of immunity is constructed and evaluated at both short timescales and at equilibrium.Results: We reveal kinetic constraints whereby variation of multiple parameters in concert leads to decreased CFR and increased pathogen fitness, whereas independent variation of the parameters decreases pathogen fitness. Smallpox, SARS-CoV-2, and influenza are analyzed as diverse representatives of human respiratory viruses. We show that highly virulent viruses, such as smallpox, are often constrained by the host behavior, whereas moderately virulent viruses, such as SARS-CoV-2, appear to be typically constrained by the relationship between the duration of immunity and CFR.Conclusions: Evolution of human respiratory epidemics appears to be often kinetically constrained and a reduction in CFR should not be assumed. These results agree with previous work demonstrating an increase in virulence for smallpox and further predict that SARS-CoV-2 is likely to continue presenting a substantial disease burden. Herd immunity against SARS-CoV-2 and viruses with similar life history traits might be unachievable without vaccination. However, partial isolation of symptomatic individuals can have a major effect on the epidemic dynamics not only by reducing the number of fatalities in the short term but also by changing the evolutionary trajectory of moderate CFR viruses towards reduced CFR.

Kinetic Constraints in the Specific Interaction between Phosphorylated Ubiquitin and Proteasomal Shuttle Factors.

Ubiquitin (Ub) specifically interacts with the Ub-associating domain (UBA) in a proteasomal shuttle factor, while the latter is involved in either proteasomal targeting or self-assembly coacervation. PINK1 phosphorylates Ub at S65 and makes Ub alternate between C-terminally relaxed (pUbRL) and retracted conformations (pUbRT). Using NMR spectroscopy, we show that pUbRL but not pUbRT preferentially interacts with the UBA from two proteasomal shuttle factors Ubqln2 and Rad23A. Yet discriminatorily, Ubqln2-UBA binds to pUb more tightly than Rad23A does and selectively enriches pUbRL upon complex formation. Further, we determine the solution structure of the complex between Ubqln2-UBA and pUbRL and uncover the thermodynamic basis for the stronger interaction. NMR kinetics analysis at different timescales further suggests an indued-fit binding mechanism for pUb-UBA interaction. Notably, at a relatively low saturation level, the dissociation rate of the UBA-pUbRL complex is comparable with the exchange rate between pUbRL and pUbRT. Thus, a kinetic constraint would dictate the interaction between Ub and UBA, thus fine-tuning the functional state of the proteasomal shuttle factors.

Modeling non-linear dielectric susceptibilities of supercooled molecular liquids.

Advances in high-precision dielectric spectroscopy have enabled access to non-linear susceptibilities of polar molecular liquids. The observed non-monotonic behavior has been claimed to provide strong support for theories of dynamic arrest based on the thermodynamic amorphous order. Here, we approach this question from the perspective of dynamic facilitation, an alternative view focusing on emergent kinetic constraints underlying the dynamic arrest of a liquid approaching its glass transition. We derive explicit expressions for the frequency-dependent higher-order dielectric susceptibilities exhibiting a non-monotonic shape, the height of which increases as temperature is lowered. We demonstrate excellent agreement with the experimental data for glycerol, challenging the idea that non-linear response functions reveal correlated relaxation in supercooled liquids.

Nanoglass and Nanocrystallization Reactions in Metallic Glasses

Strategies to change the properties of metallic glass by controlling the crystallization and the glass transition behavior are essential in promoting the application of these materials. Aside from changing the composition approaches to stabilize the glass and frustrate the nucleation and growth of crystals, new strategies at a fixed glass composition are of special interest. In this review, some recent work is summarized on new strategies to tune the properties of metallic glasses without changing composition. First, the nanocrystallization strategy is introduced that is based on the nanocrystallized microstructures such as those that develop in marginal Al-based metallic glasses. The heterogeneous and transient nucleation effects in the nanocrystallization reactions in Al-based metallic glasses are systematically investigated and can be assessed by the determination of delay time based on Flash DSC measurements. These results provide a basis to understand the strong effect of minor alloying additions on the onset of primary Al nanocrystallization and to design the novel Al-based composites with improved properties. Secondly, by an optimal annealing treatment, a liquid-cooled Au-based metallic glass can achieve very high kinetic stability to yield a large increase in glass transition temperature of 28 K and this is 3-5 times larger than the increase usually reported. The measured enthalpy decrease is about 50% of the difference between the as-cooled glass and the equilibrium crystalline state and reaches the extrapolated enthalpy of the supercooled liquid. Finally, the nano-glass strategy makes an Au-based nanoglass show ultrastable kinetic characters at low heating rate (e.g., 300 K/s) compared to a melt-spun ribbon, which is attributed to the kinetic constraint effect of nanoglobular interfaces. These results indicate that the nanoglass microstructure can act to increase metallic glass stability and provide another mechanism for the synthesis of ultrastable glass. These developments open new opportunities to improve the stability and properties and largely increase the application potentials of metallic glasses.

Hilbert space fragmentation and exact scars of generalized Fredkin spin chains

In this work, based on the Fredkin spin chain, we introduce a family of spin-$1/2$ many-body Hamiltonians with a three-site interaction featuring a fragmented Hilbert space with coexisting quantum many-body scars. The fragmentation results from an emergent kinetic constraint resembling the conserved spin configuration in the 1D Fermi-Hubbard model in the infinite onsite repulsion limit. To demonstrate the many-body scars, we construct an exact eigenstate that is in the middle of the spectrum within each fractured sub-sector displaying either logarithmic or area-law entanglement entropy. The interplay between fragmentation and scarring leads to rich tunable non-ergodic dynamics by quenching different initial states that is shown through large scale matrix product state simulations. In addition, we provide a Floquet quantum circuit that displays non-ergodic dynamics as a result of sharing the same fragmentation structure and scarring as the Hamiltonian.

Water Microdroplets Allow Spontaneously Abiotic Production of Peptides.

The chemistry of abiotic synthesis of peptides in the context of their prebiotic origins is a continuing challenge that arises from thermodynamic and kinetic constraints in aqueous media. Here we reported a strategy of microdroplets' mass spectrometry for peptide bonds formed from pure amino acids or a mixture in the presence of phosphoric acids in aqueous microdroplets. In contrast to bulk experiments, the condensation reactions proceed spontaneously under ambient conditions. The microdroplet gave a negative free-energy change (ΔG ∼ -1.1 kcal/mol), and product yields of ∼75% were obtained at the scale of a few milliseconds. Experiments in which nebulization gas pressure and external charge were varied established dependence of peptide production on the droplet size that has a high surface-to-volume ratio. It is concluded that the condensation reactions occurred at or near the air-water interfaces of microdroplets. This aqueous microdroplets approach also provides a route for chemistry synthesis in the prebiotic era.

Evolution of human respiratory virus epidemics.

Background: It is often assumed that pathogens evolve towards reduced virulence, but counterexamples abound. Faced with a new pathogen, such as SARS-CoV-2, it is highly desirable to be able to forecast the case fatality rate (CFR) and overall disease burden into the future. Considerable effort has been invested towards the development of a mathematical framework for predicting virulence evolution. Although many approaches accurately recapitulate complex outcomes, most rely on an assumed trade-off between CFR and infection rate. It is often impractical to empirically validate this constraint for human pathogens. Methods: A compartment model with parameters tuning the degree to which symptomatic individuals are isolated and the duration of immunity is constructed and evaluated at both short timescales and at equilibrium (when it exists). Results: We reveal kinetic constraints where the variation of multiple parameters in concert leads to decreased CFR and increased pathogen fitness, whereas independent variation of the parameters decreases pathogen fitness. Smallpox, SARS-CoV-2, and influenza are analyzed as diverse representatives of human respiratory viruses. We show that highly virulent viruses, such as smallpox, are likely often constrained by host behavior, whereas moderately virulent viruses, such as SARS-CoV-2, appear to be typically constrained by the relationship between the duration of immunity and CFR. Conclusions: Evolution of human respiratory epidemics appears to be often kinetically constrained and a reduction in CFR should not be assumed. Our findings imply that, without continued public health intervention, SARS-CoV-2 is likely to continue presenting a substantial disease burden. The existence of a parameter regime admitting endemic equilibrium suggests that herd immunity is unachievable. However, we demonstrate that even partial isolation of symptomatic individuals can have a major effect not only by reducing the number of fatalities in the short term but also by potentially changing the evolutionary trajectory of the virus towards reduced CFR.

Mechanisms of Carrier Density Increase of Ternary Cation-Based Amorphous Oxide Semiconductors for Thin Film Transistor Applications

Amorphous oxide semiconductor (AOS) thin film transistors (TFTs) based on In2O3 have attracted much interest for use as pixel switching elements in next generation active-matrix liquid crystal (AM-LCD) and active-matrix organic light emitting diode (AM-OLED) displays. The high field effect mobility of In2O3-based AOS devices (10–25 cm2/Vsec) offers significant performance improvements over present-day a-Si TFTs (<1 cm2/Vsec) technology. Additional advantages of AOS materials include low temperature processing (RT–300 °C), isotropic wet etch characteristics, and high optical transparency (>85 % in the visible regime) all of which make this material suitable for large area, flexible, and transparent devices on inexpensive polymer substrates.There have been considerable efforts to incorporate AOSs into devices, particularly in current-driven active matrix displays such as organic light emitting diode displays as pixel-driving switching elements. Strategies to enhance amorphous phase stability, reduce bias stress-induced threshold voltage shifts, and suppress channel carrier densities have been studied and successfully applied to the switching TFT application. Third cation elements are often added to binary cation oxide systems to limit the channel carrier generation for TFT channel application. We have recently reported that the addition of Al to InZnO, a typical binary cation material system leads to enhanced amorphous phase stability, carrier suppression capability and higher carrier mobility, up to ~45 cm2/Vs (Hall Effect mobility) and ~20 cm2/Vs (field effect mobility)1. All of these characteristics are expected to be a key enabler for realizing the next generation ultra-high-definition displays.Post-process annealing is widely employed in AOS-based TFT fabrication since annealing has been shown to improve field effect mobility2-3 and channel/metallization contacts4-5 as well as reduce trap density6, which often leads to unstable device performance or unfavorable hysteresis in their transistor characterstics6-7. However, the post-annealing is often accompanied by an increase in channel carrier density that induces an unfavorable increase in the device off-state current and operation voltages2, 8. To date, the origin of the increase in channel carrier density has not been fully understood.The current study aims to identify the origin of an increase in carrier density after low-temperature annealing conducted in air, particularly for a third-cation AOS system of InAlZnO (IAZO). Through work function investigations and bandgap analysis, the carrier density of IAZO is found to be increased by > 104 times compared to that of unannealed IAZO after low temperature annealing at 200 °C in air. Photoelectron spectroscopic studies reveal that the typical intrinsic (vacancy-based native defect) or extrinsic (cation substitution) doping mechanisms are not the primary cause of the channel carrier increase. From high pressure oxidation with much enhanced reactivity of reaction gases, it is identified that the equilibrium carrier density of IAZO is much higher than those used in typical TFT channel application. The low channel carrier density tends to increase and reach the higher equilibrium carrier density in the absence of kinetic constraints. The combinatorial investigations presented herein help understand the origin of unintentional increase in channel carrier density in amorphous oxides and its effect on the operation of TFTs.The authors gratefully acknowledge the financial supports of the U.S. NSF Award No. ECCS-1931088; the Purdue Research Foundation (Grant No. 60000029); and the Improvement of Measurement Standards and Technology for Mechanical Metrology (Grant No. 20011028) by KRISS.References Reed et al. Journal of Materials Chemistry C 2020, 8 (39), 13798-13810.Nomura et al. Applied Physics Letters 2009, 95 (1), 013502.Lee et al. Thin Solid Films 2012, 520 (10), 3764-3768.Shimura et al. Thin Solid Films 2008, 516 (17), 5899-5902.Lee et al. Journal of Applied Physics 2011, 109 (6), 063702.Ide et al. physica status solidi (a) 2019, 216 (5), 1800372.Liu et al. Journal of the American Chemical Society 2010, 132 (34), 11934-11942.Lee et al. Applied Physics Letters 2014, 104 (25), 252103.

Probing the application of kinetic theory to Mg-phyllosilicate growth with Si isotope doping

The Principle of Detailed Balance (PDB) played a defining role in the derivation of the widely-used Transition State Theory rate law equation and serves as an important link between geochemical kinetics and thermodynamics. Although significant improvements have been made in applying the PDB to comparatively simple systems (e.g., SiO2-water), experimental verification of the PDB is lacking for more complex minerals such as the phyllosilicates. Providing kinetic constraints on the rates of phyllosilicate growth from supersaturated solutions is particularly important for constructing facies models, constraining element cycling in the lacustrine and marine environments, and interpreting paleo-biogeochemistry. Here, we use 29Si isotopic doping techniques to quantify the rates of reaction between Mg-phyllosilicate substrates (amorphous Mg-silicate (a talc-like phase) and crystalline talc) and supersaturated solutions at 21 and 60 °C. The results show that the ratio of the forward and backward rates of amorphous Mg-silicate-water reaction approaches unity as the saturation state of the solution approaches the apparent solubility of amorphous Mg-silicate. The precipitation rates coupled with equivalent dissolution rates obtained from experiments with amorphous Mg-silicate substrates appear to obey the same rate function as the precipitation rates coupled with negligible dissolution rates obtained from experiments with crystalline Mg-silicate substrates over the degrees of supersaturation we explore, suggesting that the elementary step limiting the rate of precipitation remains the same. Accordingly, our results demonstrate that the PDB is applicable to amorphous Mg-phyllosilicate-water reactions, thereby reinforcing the use of TST rate equations to describe Mg-phyllosilicate growth. The experimental data can also be taken as evidence that the apparent solubility of amorphous Mg-silicate, a concept previously explained using the kinetic theory of nucleation and growth, also has a thermodynamic meaning, in that it represents a metastable equilibrium with the poorly crystalline phase. The measured, non-negligible forward and backward rates suggest that, even in this metastable state where little if any net reaction is occurring, isotopic signatures can be reset. Moreover, the significant discrepancy between the heterogeneous net growth rates on the amorphous Mg-silicate substrate versus those measured on crystalline talc and sepiolite substrates indicates that mineral crystallinity likely plays a key role in mineral growth during diagenesis.

Synthesis and Characterization of Copper Selenide and Tin Selenide Powders by Mechanical Alloying Method

This paper reports on the synthesis of copper selenide (CuSe) and tin selenide (SnSe) powders by high energy planetary ball milling, starting from elemental powders. Synthesis time and milling speed have been optimized to produce single phase CuSe and SnSe materials. The structural, compositional, morphological and optical properties of the synthesized samples have been analyzed by X-ray diffraction (XRD), transmission electron microscopy (TEM), Field Emission Scanning Electron Microscopy (FESEM) and UV-Vis. The low-temperature phase selection of the binary compound in this system is seen as a direct consequence of the thermodynamic facilitation, coupled with the capability of mechano-chemical synthesis to aid in overcoming kinetic constraints.

A Reformulation of Degree of Disequilibrium Analysis for Automatic Selection of Kinetic Constraints in the Rate-Controlled Constrained-Equilibrium Method

Abstract The rate-controlled constrained equilibrium (RCCE) is a model reduction scheme for chemical kinetics. It describes the evolution of a complex chemical system with acceptable accuracy with a number of rate controlling constraints on the associated constrained-equilibrium states of the system, much lower than the number of species in the underlying detailed kinetic model (DKM). Successful approximation of the constrained-equilibrium states requires accurate identification of the constraints. One promising procedure is the fully automatable Approximate Singular Value Decomposition of the Actual Degrees of Disequilibrium (ASVDADD) method that is capable of identifying the best constraints for a given range of thermodynamic conditions and a required level of approximation. ASVDADD is based on simple algebraic analysis of the results of the underlying DKM simulation and is focused on the behavior of the degrees of disequilibrium (DoD) of the individual chemical reactions. In this paper, we introduce an alternative ASVDADD algorithm. Unlike the original ASVDADD algorithm that require the direct computation of the DKM-derived DoDs and the identification of the set of linearly independent reactions, in the alternative algorithm, the components of the overall degree of disequilibrium vector can be computed directly by casting the DKM as an RCCE simulation considering a set of linearly independent constraints equaling the number of chemical species in size. The effectiveness and robustness of the derived constraints from the alternative procedure is examined in hydrogen/oxygen and methane/oxygen ignition delay simulations and the results are compared with those obtained from DKM.

Symmetry and conservation laws in non-holonomic mechanics

A geometric environment for the study of non-holonomic Lagrangian systems is developed. A definition of admissible displacement valid in the presence of arbitrary non-linear kinetic constraints is proposed. The meaning of ideality for non-strictly mechanical systems is analyzed. The concepts of geometric and/or dynamical symmetry of a constrained system are discussed and embodied in a subsequent non-holonomic formulation of Noether theorem. A revisitation of the results in an “extrinsic” variational language is worked out. A few examples and an appendix illustrating some properties of the manifold of admissible kinetic states are presented.

Origin of an unintended increase in carrier density of ternary cation-based amorphous oxide semiconductors

In thin film transistors (TFTs), carrier density in the channel layer is a fundamental intrinsic factor to engineer desirable TFT performance parameters such as the threshold voltage, drain current, and on-to-off ratio. Here, we report on the origin of carrier density modulation in a ternary cation system of InAlZnO (IAZO) and its effect on the TFT performance. Through work function investigations and bandgap analysis, the carrier density of IAZO is found to be increased by >104 times compared to that of unannealed IAZO after low temperature annealing at 200 °C in air. Photoelectron spectroscopic studies reveal that no significant changes were made in dopant concentrations, neither intrinsic (vacancy-based native defect) nor extrinsic (cation substitution) after annealing. From high pressure oxidation with much enhanced reactivity of reaction gases, it is identified that the equilibrium carrier density of IAZO is much higher than those used in typical TFT channel application. The low channel carrier density tends to increase and reach the higher equilibrium carrier density in the absence of kinetic constraints. This notion is further supported by a defect-state transition mechanism. The combinatorial investigations presented herein help understand the origin of the unintentional increase in channel carrier density in amorphous oxides and its effect on the operation of TFTs.

Phase Formation, Microstructure and Mechanical Properties of Mg67Ag33 as Potential Biomaterial

The phase and microstructure formation as well as mechanical properties of the rapidly solidified Mg67Ag33 (at. %) alloy were investigated. Owing to kinetic constraints effective during rapid cooling, the formation of equilibrium phases is suppressed. Instead, the microstructure is mainly composed of oversaturated hexagonal closest packed Mg-based dendrites surrounded by a mixture of phases, as probed by X-ray diffraction, electron microscopy and energy dispersive X-ray spectroscopy. A possible non-equilibrium phase diagram is suggested. Mainly because of the fine-grained dendritic and interdendritic microstructure, the material shows appreciable mechanical properties, such as a compressive yield strength and Young’s modulus of 245 ± 5 MPa and 63 ± 2 GPa, respectively. Due to this low Young’s modulus, the Mg67Ag33 alloy has potential for usage as biomaterial and challenges ahead, such as biomechanical compatibility, biodegradability and antibacterial properties are outlined.

Beyond alcohol oxidase: the methylotrophic yeast Komagataella phaffii utilizes methanol also with its native alcohol dehydrogenase Adh2.

ABSTRACTMethylotrophic yeasts are considered to use alcohol oxidases to assimilate methanol, different to bacteria which employ alcohol dehydrogenases with better energy conservation. The yeast Komagataella phaffii carries two genes coding for alcohol oxidase, AOX1 and AOX2. The deletion of the AOX1 leads to the MutS phenotype and the deletion of AOX1 and AOX2 to the Mut– phenotype. The Mut– phenotype is commonly regarded as unable to utilize methanol. In contrast to the literature, we found that the Mut– strain can consume methanol. This ability was based on the promiscuous activity of alcohol dehydrogenase Adh2, an enzyme ubiquitously found in yeast and normally responsible for ethanol consumption and production. Using 13C labeled methanol as substrate we could show that to the largest part methanol is dissimilated to CO2 and a small part is incorporated into metabolites, the biomass, and the secreted recombinant protein. Overexpression of the ADH2 gene in K. phaffii Mut– increased both the specific methanol uptake rate and recombinant protein production, even though the strain was still unable to grow. These findings imply that thermodynamic and kinetic constraints of the dehydrogenase reaction facilitated the evolution towards alcohol oxidase-based methanol metabolism in yeast.