Type-B Energetic Processes and Their Associated Scientific Implications

Recently, our work has identified two thermodynamically distinct types (A and B) of energetic processes naturally occurring on Earth. Type-A energy processes such as the classical heat engines, ATP hydrolysis, and many of the known chemical, electrical, and mechanical processes apparently follow well the second law of thermodynamics; Type-B energy processes, for example, the newly discovered thermotropic function that isothermally utilizes environmental heat energy to do useful work in driving ATP synthesis, follow the first law of thermodynamics (conservation of mass and energy) but do not have to be constrained by the second law, owing to its special asymmetric functions. In mitochondria, special asymmetric functions associated with Type-B processes comprise: 1) Transmembrane-electrostatic proton localization; 2) The transmembrane asymmetry of inner mitochondrial membrane structure with the protonic outlets of redox-driven proton-pumping protein complexes protruded away from the membrane surface by about 1–3 nm into the bulk liquid p-phase while the protonic inlet of the F0F1-ATP synthase located at the transmembrane-electrostatically localized proton (TELP) layer; and 3) The lateral asymmetry of mitochondrial cristae with an ellipsoidal shape that enhances the density of TELP at the cristae tips where the F0F1-ATP synthase enzymes are located in support of the TELP-associated thermotrophic function. The identification of Type-B energy processes indicates that there is an entirely new world of physical and energy sciences yet to be fully explored. Innovative efforts exploring Type-B processes to enable isothermally utilizing endless environmental heat energy could help liberate all people from their dependence on fossil fuel energy, thus helping to reduce greenhouse gas CO2 emissions and control climate change, with the goal of a sustainable future for humanity on Earth

Protocol Measuring Horizontal Gene Transfer From Algae to Non-Photosynthetic Organisms

Horizontal gene transfer (HGT) is a natural process for an organism to transfer genetic material to another organism that is a completely different species, for example, from a blue-green alga to a non-photosynthetic bacterium. The phenomenon of HGT is not only of an interest to the science of molecular genetics and biology, but also to the biosafety issue of genetic engineering. The novel protocol reported here for the first time teaches how to measure HGT from a genetically engineered (GE) blue-green alga (gene donor) to wild-type E. coli (recipient). This novel protocol can be used to measure HGT frequency for both plasmid transgenes and/or genomic transgenes from a donor to recipient organism. •According to this novel protocol, the HGT frequency may be calculated from the number of HGT recipient colonies observed, the number of recipient cells plated, and the donor-recipient co-incubation time. •This approach can also help test the possible HGT routes to assess whether a HGT is through a direct cell-to-cell interaction or by an indirect cell-to-liquid environment-to-cell process. •The protocol may be applied in full and/or in part with adjustments to measure HGT for a wide range of donor and recipient organisms of interest

Electrostatically Localized Proton Bioenergetics: Better Understanding Membrane Potential

In Mitchell\u27s chemiosmotic theory, membrane potential Δψ was given as the electric potential difference across the membrane. However, its physical origin for membrane potential Δψ was not well explained. Using the Lee proton electrostatic localization model with a newly formulated equation for protonic motive force (pmf) that takes electrostatically localized protons into account, membrane potential has now been better understood as the voltage difference contributed by the localized surface charge density ([H-+L] + nΣ i=1 [M(i+)L]) at the liquid-membrane interface as in an electrostatically localized protons/cations-membrane-anions capacitor. That is, the origin of membrane potential Δψ is now better understood as the electrostatic formation of the localized surface charge density that is the sum of the electrostatically localized proton concentration [H+L ] and the localized non-proton cations density nΣ(i=1) [M(i+)L] at the liquid membrane interface. The total localized surface charge density equals to the ideal localized proton population density [H+L]0 before the cation-proton exchange process; since the cation proton exchange process does not change the total localized charges density, neither does it change to the membrane potential Δψ. The localized proton concentration [H+L] represents the dominant component, which accounts about 78% of the total localized surface charge density at the cation-proton exchange equilibrium state in animal mitochondria. Liquid water as a protonic conductor may play a significant role in the biological activities of membrane potential formation and utilization

Isothermal Environmental Heat Energy Utilization by Transmembrane Electrostatically Localized Protons at the Liquid-Membrane Interface

This study employing the latest theory on transmembrane electrostatic proton localization has now, for the first time, consistently elucidated a decades-longstanding bioenergetic conundrum in alkalophilic bacteria and more importantly discovered an entirely new feature: isothermal environmental heat utilization by electrostatically localized protons at the liquid-membrane interface. It was surprisingly revealed that the protonic motive force (equivalent to Gibbs free energy) from the isothermal environmental heat energy utilization through the electrostatically localized protons is not constrained by the overall energetics of the redox-driven proton pump system because of the following: (a) the transmembrane electrostatically localized protons are not free to move away from the membrane surface as a protonic capacitor feature; (b) the proton pumps embedded in the cell membrane extend beyond the localized proton layer apparently as an asymmetric property of the biological membrane; and (c) the protonic inlet mouth of the ATP synthase that accepts protons is located within this layer as another natural property of the asymmetric biological membrane. This work has now, for the first time, shown a novel thermotrophic feature where biological systems can isothermally utilize environmental heat energy through transmembrane electrostatically localized protons to help drive ATP synthesis

Protonic Capacitor: Elucidating the Biological Significance of Mitochondrial Cristae Formation

For decades, it was not entirely clear why mitochondria develop cristae? The work employing the transmembrane-electrostatic proton localization theory reported here has now provided a clear answer to this fundamental question. Surprisingly, the transmembrane-electrostatically localized proton concentration at a curved mitochondrial crista tip can be significantly higher than that at the relatively flat membrane plane regions where the proton-pumping respiratory supercomplexes are situated. The biological significance for mitochondrial cristae has now, for the first time, been elucidated at a protonic bioenergetics level: 1) The formation of cristae creates more mitochondrial inner membrane surface area and thus more protonic capacitance for transmembrane-electrostatically localized proton energy storage; and 2) The geometric effect of a mitochondrial crista enhances the transmembraneelectrostatically localized proton density to the crista tip where the ATP synthase can readily utilize the localized proton density to drive ATP synthesis

Energy Renewal: Isothermal Utilization of Environmental Heat Energy with Asymmetric Structures

Through the research presented herein, it is quite clear that there are two thermodynamically distinct types (A and B) of energetic processes naturally occurring on Earth. Type A, such as glycolysis and the tricarboxylic acid cycle, apparently follows the second law well; Type B, as exemplified by the thermotrophic function with transmembrane electrostatically localized protons presented here, does not necessarily have to be constrained by the second law, owing to its special asymmetric function. This study now, for the first time, numerically shows that transmembrane electrostatic proton localization (Type-B process) represents a negative entropy event with a local protonic entropy change (Δ SL) in a range from −95 to −110 J/K∙mol. This explains the relationship between both the local protonic entropy change (ΔSL) and the mitochondrial environmental temperature (T) and the local protonic Gibbs free energy (ΔGL=TΔSL) in isothermal environmental heat utilization. The energy efficiency for the utilization of total protonic Gibbs free energy (ΔGT including ΔGL=TΔSL) in driving the synthesis of ATP is estimated to be about 60%, indicating that a significant fraction of the environmental heat energy associated with the thermal motion kinetic energy (kBT) of transmembrane electrostatically localized protons is locked into the chemical form of energy in ATP molecules. Fundamentally, it is the combination of water as a protonic conductor, and thus the formation of protonic membrane capacitor, with asymmetric structures of mitochondrial membrane and cristae that makes this amazing thermotrophic feature possible. The discovery of energy Type-B processes has inspired an invention (WO 2019/136037 A1) for energy renewal through isothermal environmental heat energy utilization with an asymmetric electron-gated function to generate electricity, which has the potential to power electronic devices forever, including mobile phones and laptops. This invention, as an innovative Type-B mimic, may have many possible industrial applications and is likely to be transformative in energy science and technologies for sustainability on Earth

Proton-Electrostatic Localization: Explaining the Bioenergetic Conundrum in Alkalophilic Bacteria

The decades-longstanding energetic conundrum of alkalophilic bacteria as to how they are able to synthesize ATP has now, for the first time, been clearly solved using the proton-electrostatics localization hypothesis. This is a major breakthrough advance in understanding proton-coupling bioenergetics over the Nobel-prize work of Peter Mitchell’s chemiosmotic theory. The widespread textbook Mitchellian proton motive force (pmf) equation has now been significantly revised. Use of the newly derived equation results in an overall pmf value (215~233 mV) that is more than 4 times larger than that (44.3 mV) calculated from the Mitchellian equation for the alkalophilic bacteria growing at pH 10.5. This newly calculated value is sufficient to overcome the observed phosphorylation potential ΔGp of −478 mV to synthesize ATP in the bacteria, which can now explain the 30-year-longstanding bioenergetics conundrum. This finding may have fundamental implications not only in the science of bioenergetics but also in understanding the importance of water to life not only as a solvent and substrate but also as a proton conductor for proton coupling energy transduction

TELP Theory: Elucidating the Major Observations of Rieger et al. 2021 in Mitochondria

The transmembrane-electrostatically localized protons (TELP) theory may represent a complementary development to Mitchell\u27s chemiosmotic theory. The combination of the two together can now excellently explain the energetics in mitochondria. Our calculated transmembrane-attractive force between an excess proton and an excess hydroxide explains how TELP may stay within a 1-nm thin layer at the liquid-membrane interface. Consequently, any pH sensor (sEcGFP) located at least 2–3 nm away from the membrane surface will not be able to see TELP. This feature as predicted from the TELP model was observed exactly in the experiment of Rieger et al., 2021. In contrast to their belief “the Δp at ATP synthase is almost negligible under OXPHOS conditions”, I find, when TELP activity is included in the energy calculations, there is plenty of total protonic Gibbs free energy (ΔGT) well above the physiologically required value of −24.5 kJ mol−1 to drive ATP synthesis through F0F1-ATP synthase

Mitochondrial Energetics with Transmembrane Electrostatically Localized Protons: Do We Have a Thermotrophic Feature?

Transmembrane electrostatically localized protons (TELP) theory has been recently recognized as an important addition over the classic Mitchell’s chemiosmosis; thus, the proton motive force (pmf) is largely contributed from TELP near the membrane. As an extension to this theory, a novel phenomenon of mitochondrial thermotrophic function is now characterized by biophysical analyses of pmf in relation to the TELP concentrations at the liquid-membrane interface. This leads to the conclusion that the oxidative phosphorylation also utilizes environmental heat energy associated with the thermal kinetic energy (kBT) of TELP in mitochondria. The local pmf is now calculated to be in a range from 300 to 340 mV while the classic pmf (which underestimates the total pmf) is in a range from 60 to 210 mV in relation to a range of membrane potentials from 50 to 200 mV. Depending on TELP concentrations in mitochondria, this thermotrophic function raises pmf significantly by a factor of 2.6 to sixfold over the classic pmf. Therefore, mitochondria are capable of effectively utilizing the environmental heat energy with TELP for the synthesis of ATP, i.e., it can lock heat energy into the chemical form of energy for cellular functions

Ozonized Biochar Filtrate Effects on the Growth of \u3ci\u3ePseudomonas putida\u3c/i\u3e and Cyanobacteria \u3ci\u3eSynechococcus elongatus\u3c/i\u3e PCC 7942

Background Biochar ozonization was previously shown to dramatically increase its cation exchange capacity, thus improving its nutrient retention capacity. The potential soil application of ozonized biochar warrants the need for a toxicity study that investigates its effects on microorganisms. Results In the study presented here, we found that the filtrates collected from ozonized pine 400 biochar and ozonized rogue biochar did not have any inhibitory effects on the soil environmental bacteria Pseudomonas putida, even at high dissolved organic carbon (DOC) concentrations of 300 ppm. However, the growth of Synechococcus elongatus PCC 7942 was inhibited by the ozonized biochar filtrates at DOC concentrations greater than 75 ppm. Further tests showed the presence of some potential inhibitory compounds (terephthalic acid and p-toluic acid) in the filtrate of non-ozonized pine 400 biochar; these compounds were greatly reduced upon wet-ozonization of the biochar material. Nutrient detection tests also showed that dry-ozonization of rogue biochar enhanced the availability of nitrate and phosphate in its filtrate, a property that may be desirable for soil application. Conclusion Ozonized biochar substances can support soil environmental bacterium Pseudomonas putida growth, since ozonization detoxifies the potential inhibitory aromatic molecules