Biocalorimetry-aided monitoring of fungal pretreatment of lignocellulosic agricultural residues.

The present study aimed to investigate whether and how non-invasive biocalorimetric measurements could serve for process monitoring of fungal pretreatment during solid-state fermentation (SSF) of lignocellulosic agricultural residues such as wheat straw. Seven filamentous fungi representing different lignocellulose decay types were employed. Water-soluble sugars being immediately available after fungal pretreatment and those becoming water-extractable after enzymatic digestion of pretreated wheat straw with hydrolysing (hemi)cellulases were considered to constitute the total bioaccessible sugar fraction. The latter was used to indicate the success of pretreatments and linked to corresponding species-specific metabolic heat yield coefficients (YQ/X) derived from metabolic heat flux measurements during fungal wheat straw colonisation. An YQ/X range of about 120 to 140 kJ/g was seemingly optimal for pretreatment upon consideration of all investigated fungi and application of a non-linear Gaussian fitting model. Upon exclusion from analysis of the brown-rot basidiomycete Gloeophyllum trabeum, which differs from all other here investigated fungi in employing extracellular Fenton chemistry for lignocellulose decomposition, a linear relationship where amounts of total bioaccessible sugars were suggested to increase with increasing YQ/X values was obtained. It remains to be elucidated whether an YQ/X range being optimal for fungal pretreatment could firmly be established, or if the sugar accessibility for post-treatment generally increases with increasing YQ/X values as long as "conventional" enzymatic, i.e. (hemi)cellulase-based, lignocellulose decomposition mechanisms are operative. In any case, metabolic heat measurement-derived parameters such as YQ/X values may become very valuable tools supporting the assessment of the suitability of different fungal species for pretreatment of lignocellulosic substrates. KEY POINTS: • Biocalorimetry was used to monitor wheat straw pretreatment with seven filamentous fungi. • Metabolic heat yield coefficients (YQ/X) seem to indicate pretreatment success. • YQ/X values may support the selection of suitable fungal strains for pretreatment.

Enhancing insights: exploring the information content of calorespirometric ratio in dynamic soil microbial growth processes through calorimetry.

Catalytic activity of microbial communities maintains the services and functions of soils. Microbial communities require energy and carbon for microbial growth, which they obtain by transforming organic matter (OM), oxidizing a fraction of it and transferring the electrons to various terminal acceptors. Quantifying the relations between matter and energy fluxes is possible when key parameters such as reaction enthalpy (∆rH), energy use efficiency (related to enthalpy) (EUE), carbon use efficiency (CUE), calorespirometric ratio (CR), carbon dioxide evolution rate (CER), and the apparent specific growth rate (µapp) are known. However, the determination of these parameters suffers from unsatisfying accuracy at the technical (sample size, instrument sensitivity), experimental (sample aeration) and data processing levels thus affecting the precise quantification of relationships between carbon and energy fluxes. To address these questions under controlled conditions, we analyzed microbial turnover processes in a model soil amended using a readily metabolizable substrate (glucose) and three commercial isothermal microcalorimeters (MC-Cal/100P, TAM Air and TAM III) with different sample sizes meaning varying volume-related thermal detection limits (LODv) (0.05-1mW L-1). We conducted aeration experiments (aerated and un-aerated calorimetric ampoules) to investigate the influence of oxygen limitation and thermal perturbation on the measurement signal. We monitored the CER by measuring the additional heat caused by CO2 absorption using a NaOH solution acting as a CO2 trap. The range of errors associated with the calorimetrically derived µapp, EUE, and CR was determined and compared with the requirements for quantifying CUE and the degree of anaerobicity (ηA). Calorimetrically derived µapp and EUE were independent of the instrument used. However, instruments with a low LODv yielded the most accurate results. Opening and closing the ampoules for oxygen and CO2 exchange did not significantly affect metabolic heats. However, regular opening during calorimetrically derived CER measurements caused significant measuring errors due to strong thermal perturbation of the measurement signal. Comparisons between experimentally determined CR, CUE,ηA, and modeling indicate that the evaluation of CR should be performed with caution.

Core-Shell Polydopamine/Cu Nanometer Rods Efficiently Deactivate Microbes by Mimicking Chloride-Activated Peroxidases.

Cu-modified nanoparticles have been designed to mimic peroxidase, and their potent antibacterial and anti-biofilm abilities have been widely investigated. In this study, novel core-shell polydopamine (PDA)/Cu4(OH)6SO4 crystal (PDA/Cu) nanometer rods were prepared. The PDA/Cu nanometer rods show similar kinetic behaviors to chloride-activated peroxidases, exhibit excellent photothermal properties, and are sensitive to the concentrations of pH values and the substrate (i.e., H2O2). PDA/Cu nanometer rods could adhere to the bacteria and catalyze hydrogen peroxide (H2O2) to generate more reactive hydroxy radicals (•OH) against Staphylococcus aureus and Escherichia coli, Furthermore, PDA/Cu nanometer rods show enhanced catalytic and photothermal synergistic antibacterial activity. This work provides a simple, inexpensive, and effective strategy for antibacterial applications.

Fungal Lignocellulose Utilisation Strategies from a Bioenergetic Perspective: Quantification of Related Functional Traits Using Biocalorimetry.

In the present study, we investigated whether a non-invasive metabolic heat flux analysis could serve the determination of the functional traits in free-living saprotrophic decomposer fungi and aid the prediction of fungal influences on ecosystem processes. For this, seven fungi, including ascomycete, basidiomycete, and zygomycete species, were investigated in a standardised laboratory environment, employing wheat straw as a globally relevant lignocellulosic substrate. Our study demonstrates that biocalorimetry can be employed successfully to determine growth-related fungal activity parameters, such as apparent maximum growth rates (AMGR), cultivation times until the observable onset of fungal growth at AMGR (tAMGR), quotients formed from the AMGR and tAMGR (herein referred to as competitive growth potential, CGP), and heat yield coefficients (YQ/X), the latter indicating the degree of resource investment into fungal biomass versus other functional attributes. These parameters seem suitable to link fungal potentials for biomass production to corresponding ecological strategies employed during resource utilisation, and therefore may be considered as fungal life history traits. A close connection exists between the CGP and YQ/X values, which suggests an interpretation that relates to fungal life history strategies.

Applicability and information value of biocalorimetry for the monitoring of fungal solid-state fermentation of lignocellulosic agricultural by-products.

The applicability of biocalorimetry for monitoring fungal conversion of lignocellulosic agricultural by-products during solid-state fermentation (SSF) was substantiated through linking the non-invasive measurement of metabolic heat fluxes to conventional invasive determination of fungal activity (growth, substrate degradation, enzyme activity) parameters. For this, the fast-growing, cellulose-utilising ascomycete Stachybotrys chlorohalonata and the comparatively slow-growing litter-decay basidiomycete Stropharia rugosoannulata were investigated as model organisms during growth on solid wheat straw. Both biocalorimetric and non-calorimetric data may suggest R (ruderal)- and C (combative)-selected life history strategies in S. chlorohalonata and S. rugosoannulata, respectively. For both species, a strong linear correlation of the released metabolic heat with the corresponding fungal biomass was observed. Species-specific YQ/X values (metabolic heat released per fungal biomass unit) were obtained, which potentially enable use of biocalorimetric signals for the quantification of fungal biomass during single-species SSF processes. Moreover, YQ/X values may also indicate different fungal life history strategies and therefore be considered as useful parameters aiding fungal ecology research.

Enzymatic degradation of polyethylene terephthalate nanoplastics analyzed in real time by isothermal titration calorimetry.

Plastics are globally used for a variety of benefits. As a consequence of poor recycling or reuse, improperly disposed plastic waste accumulates in terrestrial and aquatic ecosystems to a considerable extent. Large plastic waste items become fragmented to small particles through mechanical and (photo)chemical processes. Particles with sizes ranging from millimeter (microplastics, <5 mm) to nanometer (nanoplastics, NP, <100 nm) are apparently persistent and have adverse effects on ecosystems and human health. Current research therefore focuses on whether and to what extent microorganisms or enzymes can degrade these NP. In this study, we addressed the question of what information isothermal titration calorimetry, which tracks the heat of reaction of the chain scission of a polyester, can provide about the kinetics and completeness of the degradation process. The majority of the heat represents the cleavage energy of the ester bonds in polymer backbones providing real-time kinetic information. Calorimetry operates even in complex matrices. Using the example of the cutinase-catalyzed degradation of polyethylene terephthalate (PET) nanoparticles, we found that calorimetry (isothermal titration calorimetry-ITC) in combination with thermokinetic models is excellently suited for an in-depth analysis of the degradation processes of NP. For instance, we can separately quantify i) the enthalpy of surface adsorption ∆AdsH = 129 ± 2 kJ mol-1, ii) the enthalpy of the cleavage of the ester bonds ∆EBH = -58 ± 1.9 kJ mol-1 and the apparent equilibrium constant of the enzyme substrate complex K = 0.046 ± 0.015 g L-1. It could be determined that the heat production of PET NP degradation depends to 95% on the reaction heat and only to 5% on the adsorption heat. The fact that the percentage of cleaved ester bonds (η = 12.9 ± 2.4%) is quantifiable with the new method is of particular practical importance. The new method promises a quantification of enzymatic and microbial adsorption to NP and their degradation in mimicked real-world aquatic conditions.

New thermodynamic activity-based approach allows predicting the feasibility of glycolysis.

Thermodynamic feasibility analyses help evaluating the feasibility of metabolic pathways. This is an important information used to develop new biotechnological processes and to understand metabolic processes in cells. However, literature standard data are uncertain for most biochemical reactions yielding wrong statements concerning their feasibility. In this article we present activity-based equilibrium constants for all the ten glycolytic reactions, accompanied by the standard reaction data (standard Gibbs energy of reaction and standard enthalpy of reaction). We further developed a thermodynamic activity-based approach that allows to correctly determine the feasibility of glycolysis under different chosen conditions. The results show for the first time that the feasibility of glycolysis can be explained by thermodynamics only if (1) correct standard data are used and if (2) the conditions in the cell at non-equilibrium states are accounted for in the analyses. The results here will help to determine the feasibility of other metabolisms and to understand metabolic processes in cells in the future.

Thermodynamics and Kinetics of Glycolytic Reactions. Part I: Kinetic Modeling Based on Irreversible Thermodynamics and Validation by Calorimetry.

In systems biology, material balances, kinetic models, and thermodynamic boundary conditions are increasingly used for metabolic network analysis. It is remarkable that the reversibility of enzyme-catalyzed reactions and the influence of cytosolic conditions are often neglected in kinetic models. In fact, enzyme-catalyzed reactions in numerous metabolic pathways such as in glycolysis are often reversible, i.e., they only proceed until an equilibrium state is reached and not until the substrate is completely consumed. Here, we propose the use of irreversible thermodynamics to describe the kinetic approximation to the equilibrium state in a consistent way with very few adjustable parameters. Using a flux-force approach allowed describing the influence of cytosolic conditions on the kinetics by only one single parameter. The approach was applied to reaction steps 2 and 9 of glycolysis (i.e., the phosphoglucose isomerase reaction from glucose 6-phosphate to fructose 6-phosphate and the enolase-catalyzed reaction from 2-phosphoglycerate to phosphoenolpyruvate and water). The temperature dependence of the kinetic parameter fulfills the Arrhenius relation and the derived activation energies are plausible. All the data obtained in this work were measured efficiently and accurately by means of isothermal titration calorimetry (ITC). The combination of calorimetric monitoring with simple flux-force relations has the potential for adequate consideration of cytosolic conditions in a simple manner.

Thermodynamics and Kinetics of Glycolytic Reactions. Part II: Influence of Cytosolic Conditions on Thermodynamic State Variables and Kinetic Parameters.

For systems biology, it is important to describe the kinetic and thermodynamic properties of enzyme-catalyzed reactions and reaction cascades quantitatively under conditions prevailing in the cytoplasm. While in part I kinetic models based on irreversible thermodynamics were tested, here in part II, the influence of the presumably most important cytosolic factors was investigated using two glycolytic reactions (i.e., the phosphoglucose isomerase reaction (PGI) with a uni-uni-mechanism and the enolase reaction with an uni-bi-mechanism) as examples. Crowding by macromolecules was simulated using polyethylene glycol (PEG) and bovine serum albumin (BSA). The reactions were monitored calorimetrically and the equilibrium concentrations were evaluated using the equation of state ePC-SAFT. The pH and the crowding agents had the greatest influence on the reaction enthalpy change. Two kinetic models based on irreversible thermodynamics (i.e., single parameter flux-force and two-parameter Noor model) were applied to investigate the influence of cytosolic conditions. The flux-force model describes the influence of cytosolic conditions on reaction kinetics best. Concentrations of magnesium ions and crowding agents had the greatest influence, while temperature and pH-value had a medium influence on the kinetic parameters. With this contribution, we show that the interplay of thermodynamic modeling and calorimetric process monitoring allows a fast and reliable quantification of the influence of cytosolic conditions on kinetic and thermodynamic parameters.

Influence of cytosolic conditions on the reaction equilibrium and the reaction enthalpy of the enolase reaction accessed by calorimetry and van 't HOFF.

BACKGROUND: Thermodynamic methods are finding more and more applications in systems biology, which attempts to understand cell functions mechanistically. Unfortunately, the state variables used (reaction enthalpy and Gibbs energy) do not take sufficient account of the conditions inside of cells, especially the crowding with macromolecules. METHODS: For this reason, the influence of crowding agents and various other parameters such as salt concentrations, pH and temperature on equilibrium position and reaction enthalpy of the glycolytic example reaction 9 (2-Phospoglycerate - > Phosphoenolpyruvate + H2O) was investigated. The conditions were chosen to be as close as possible to the cytosolic conditions. Poly(ethylene glycol) MW = 20,000 g mol-1 (PEG 20,000) was used to analyze the influence of crowding with macromolecules. The equation of state electrolyte Perturbed-Chain Statistical Associating Fluid Theory (ePC-SAFT) was applied to consider the influence of crowding agents on the reaction equilibria. RESULTS AND CONCLUSIONS: For the reaction enthalpies and for the equilibria, it was found that the influence of salts and temperature is not pronounced while that of pH and PEG 20,000 concentration is considerable. Furthermore, it could be shown that under identical measurement conditions there are no differences between the van 't Hoff and the calorimetrically determined reaction enthalpy. GENERAL SIGNIFICANCE: The results show how important it is to consider the special cytosolic conditions when applying thermodynamic data in systems biology.

Rapid culture-based detection of Legionella pneumophila using isothermal microcalorimetry with an improved evaluation method.

The detection and quantification of Legionella pneumophila (responsible for legionnaire's disease) in water samples can be achieved by various methods. However, the culture-based ISO 11731:2017, which is based on counts of colony-forming units per ml (CFU·ml-1 ) is still the gold standard for quantification of Legionella species (spp.). As a powerful alternative, we propose real-time monitoring of the growth of L. pneumophila using an isothermal microcalorimeter (IMC). Our results demonstrate that, depending on the initial concentration of L. pneumophila, detection times of 24-48 h can be reliably achieved. IMC may, therefore, be used as an early warning system for L. pneumophila contamination. By replacing only visual detection of growth by a thermal sensor, but otherwise maintaining the standardized protocol of the ISO 11731:2017, the new procedure could easily be incorporated into existing standards. The exact determination of the beginning of metabolic heat is often very difficult because at the beginning of the calorimetric signal the thermal stabilization and the metabolic heat development overlap. Here, we propose a new data evaluation based on the first derivation of the heat flow signal. The improved evaluation method can further reduce detection times and significantly increase the reliability of the IMC approach.

Standard Gibbs energy of metabolic reactions: V. Enolase reaction.

The glycolytic pathway is one of the most important pathways for living organisms, due to its role in energy production and as supplier of precursors for biosynthesis in living cells. This work focuses on determination of the standard Gibbs energy of reaction ΔRg'0 of the enolase reaction, the ninth reaction in the glycolysis pathway. Exact ΔRg'0 values are required to predict the thermodynamic feasibility of single metabolic reactions or even of metabolic reaction sequences under cytosolic conditions. So-called "apparent" standard data from literature are only valid at specific conditions. Nevertheless, such data are often used in pathway analyses, which might lead to misinterpretation of the results. In this work, equilibrium measurements were combined with activity coefficients in order to obtain new standard values ΔRg'0 for the enolase reaction that are independent of the cytosolic conditions. Reaction equilibria were measured at different initial substrate concentrations and temperatures of 298.15 K, 305.15 K and 310.15 K at pH 7. The activity coefficients were predicted using the equation of state electrolyte Perturbed-Chain Statistical Associating Fluid Theory (ePC-SAFT). The ePC-SAFT parameters were taken from literature or fitted to new experimentally determined osmotic coefficients and densities. At 298.15 K and pH 7, a ΔRg'0(298.15 K, pH 7) value of -2.8 ± 0.2 kJ mol-1 was obtained. This value differs by up to 5 kJ mol-1 from literature data. Reasons are the poorly defined "standard" conditions and partly undefined reaction conditions of literature works. Finally, using temperature-dependent equilibrium constants and the van 't Hoff equation, the standard enthalpy of reaction of ΔRh'0(298.15 K, pH 7) = 27 ± 10 kJ mol-1 was determined, and a similar value was found by quantum-chemistry calculations.

Rapid Calorimetric Detection of Bacterial Contamination: Influence of the Cultivation Technique.

Modern isothermal microcalorimeters (IMC) are able to detect the metabolic heat of bacteria with an accuracy sufficient to recognize even the smallest traces of bacterial contamination of water, food, and medical samples. The modern IMC techniques are often superior to conventional detection methods in terms of the detection time, reliability, labor, and technical effort. What is missing is a systematic analysis of the influence of the cultivation conditions on calorimetric detection. For the acceptance of IMC techniques, it is advantageous if already standardized cultivation techniques can be combined with calorimetry. Here we performed such a systematic analysis using Lactobacillus plantarum as a model bacterium. Independent of the cultivation techniques, IMC detections were much faster for high bacterial concentrations (>102 CFU⋅mL-1) than visual detections. At low bacterial concentrations (<102 CFU⋅mL-1), detection times were approximately the same. Our data demonstrate that all kinds of traditional cultivation techniques like growth on agar (GOA) or in agar (GIA), in liquid media (GL) or on agar after enrichment via membrane filtration (GF) can be combined with IMC. The order of the detection times was GF < GIA ≈ GL ≈ GOA. The observed linear relationship between the calorimetric detection times and the initial bacterial concentrations can be used to quantify the bacterial contamination. Further investigations regarding the correlation between the filling level (in mm) of the calorimetric vessel and the specific maximum heat flow (in µW⋅g-1) illustrated two completely different results for liquid and solid media. Due to the better availability of substrates and the homogeneous distribution of bacteria growing in a liquid medium, the volume-related maximum heat flow was independent on the filling level of the calorimetric vessels. However, in a solid medium, the volume-related maximum heat flow approached a threshold and achieved a maximum at low filling levels. This fundamentally different behavior can be explained by the spatial limitation of the growth of bacterial colonies and the reduced substrate supply due to diffusion.

Photocalorespirometry (Photo-CR): A Novel Method for Access to Photosynthetic Energy Conversion Efficiency.

One key parameter for assessing the CO2 fixation in aquatic ecosystems but also for the productivity of photobioreactors is the energy conversion efficiency (PE) by the photosynthetic apparatus. PE strictly depends on a range of different fluctuating environmental conditions and is therefore highly variable. PE is the result of complex metabolic control. At the moment PE can only be determined indirectly. Furthermore, the currently available techniques either capture only short time processes, thus reflecting only parts of the photosynthetic engine, or quantify the total process but only with limited time resolution. To close this gap, we suggest for the first time the direct measurement of the fixed energy combined with respirometry, called photocalorespirometry (Photo-CR). The proof of the principle of Photo-CR was established with the microalga Chlamydomonas reinhardtii. The simultaneous measurement of oxygen production and energy fixation provides an calorespirometric ratio of -(437.9 ± 0.7) kJ mol-1 under low light conditions. The elevated calorespirometric ratio under high light conditions provides an indication of photo-protective mechanisms. The Photo-CR delivers the PE in real time, depending on the light intensity. Energetic differences less than 0.14% at radiation densities of up to 800 µE m-2 s-1 can be quantified. Other photosynthetic growth parameters (e.g. the specific growth rate of 0.071 h-1, the cell specific energy conservation of 30.9 ± 1.3 pW cell-1 at 150 µE m-2 s-1 and the number of photons (86.8) required to fix one molecule of CO2) can easily be derived from the Photo-CR data.

High-frequency, dielectric spectroscopy for the detection of electrophysiological/biophysical differences in different bacteria types and concentrations.

This paper describes a novel technique to quantify and identify bacterial cultures of Bacillus Subtilis (2.10-1.30 × 109 CFU mL-1) and Escherichia Coli (1.60-1.00 × 109 CFU mL-1), in corn oil using dielectric spectroscopy at elevated frequencies of 0.0100-20.0 GHz. This technique is using the electrophysiological/biophysical differences (e.g. gram positive and gram negative) between various bacteria types, as a basis to distinguish between bacteria concentrations and bacteria types. A close-ended, coaxial probe of 20.0 mm long sample-holder was developed and used to calculate the dielectric constant from the measured S parameters of the bacterial cultures, using the Nicolson-Ross-Weir method. This technique shows a linear relationship (r2 ≥ 0.999) between the dielectric constant and the cell concentration, at 16.0 GHz. The sensitivity of the technique is 0.177 × 109 (CFU mL-1)-1 for B. Subtilis (with a size of 10.0 × 1.00 µm), 0.322 × 109 (CFU mL-1)-1 for E. Coli (with a size of 2.00 × 0.500 µm) and 0.913 × 109 (CFU mL-1) -1 for their 1:1 mixture, while the response time is 60.0s. The dependency of dielectric constant on the bacterial cell concentration at a given frequency can be potentially exploited for measuring bacterial concentrations and biophysical differences.

A fast and reliable method for monitoring of prophage-activating chemicals.

Bacteriophages, that is viruses that infect bacteria, either lyse bacteria directly or integrate their genome into the bacterial genome as so-called prophages, where they remain at a silent state. Both phages and bacteria are able to survive in this state. However, prophages can be reactivated with the introduction of chemicals, followed by the release of a high number of phage particles, which could infect other bacteria, thus harming ecosystems by a viral bloom. The basics for a fast, automatable analytical method for the detection of prophage-activating chemicals are developed and successfully tested here. The method exploits the differences in metabolic heat produced by Escherichia coli with (λ+) and without the lambda prophages (λ-). Since the metabolic heat primarily reflects opposing effects (i.e. the reduction of heat-producing cells by lysis and enhanced heat production to deliver the energetic costs for the synthesis of phages), a systematic analysis of the influence of the different conditions (experimentally and in silico) was performed and revealed anoxic conditions to be best suited. The main advantages of the suggested monitoring method are not only the possibility of obtaining fast results (after only few hours), but also the option for automation, the low workload (requires only few minutes) and the suitability of using commercially available instruments. The future challenge following this proof of principle is the development of thermal transducers which allow for the electronic subtraction of the λ+ from the λ- signal.

An enhanced bioindicator for calorimetric monitoring of prophage-activating chemicals in the trace concentration range.

Viruses that infect bacteria (bacteriophages) can either lyse bacteria directly or integrate their genome into the bacterial genome. In the latter case, the viral genome (called prophage) remains dormant, and both phages and bacteria are able to survive in this state. But the silent prophages can be reactivated by, e.g., chemicals, accompanied by the release of substantial quantities of phage particles that further infect other phage-sensitive bacteria, thus harming ecosystems or technical systems by way of a viral bloom. Recently, a calorimetric method was developed to monitor the prophage-activating properties of chemicals. The method evaluates the difference in the metabolic heat of the Escherichia coli bioindicator with (λ+) and without (λ-) lambda prophages under the influence of the test substances. Simulations and experiments clearly demonstrate that the sensitivity of the test can be significantly improved, when a customized mixture of λ+ and λ- E. coli strains is used for enhanced bioindication. Hence, the new method mirrors a common situation in nature, where bacteria with and without prophages coexist. In summary, a monitoring method is suggested that provides quick results (after few hours) and offers both the option for automation with low workload (requires only a few minutes) and usage of commercially available instruments.

Mn-Doped ZnSe quantum dots initiated mild and rapid cation exchange for tailoring the composition and optical properties of colloid nanocrystals: novel template, new applications.

Although cation exchange (CE) has been studied for many years and some mechanisms were proposed, there is still a knowledge gap in CE and problems such as the need for high temperature and it being time-consuming are still unaddressed. We developed a new mild strategy for CE by introducing a new ideal template and first applied this doping strategy to detect Cd2+ and Hg2+. This strategy adopted Mn-doped ZnSe quantum dots (QDs) as the template and the introduction occurs via a two-step CE reaction: first Zn2+ was partially substituted by X (X = Cd2+, Hg2+, Cu2+, Ag+ or Pb2+), later Mn2+ (in the deep structure of QDs) was substituted by X. Remarkably, Mn2+ in the lattice can be easily substituted by a dopant and its replacement by a dopant helps to bury the metal ions. The ultra-fast introduction of Cd2+ and Hg2+ (70 minutes for Cd2+ and 19 minutes for Hg2+) was realized at room temperature; other metal ions such as Ag+, Cu2+ and Pb2+ can be buried at 50 °C. This mild reaction temperature offers a solution for introducing impurities without sacrificing the interfacial structure of nanocrystals. HRTEM, XPS and ICP measurements were applied to analyze the introduction process. Furthermore, the spectroscopic method was employed to analyze the introduction, migration and distribution of metal ions. Then, we proposed a mechanism for the chemical conversion of nanocrystals by CE. Through this strategy, full-color light-emitting doped QDs were fabricated. Strikingly, a new turn-on probe for the detection of Cd2+ and Hg2+ with improved selectivity was developed by adopting this doping strategy. The detection limit is 36 nM for Cd2+ and 20 nM for Hg2+, which is competitive with the limit of detection reported by other groups using QDs as sensors.