A process for the preparation of a specific carrier immobilized multifunctional microbial consortium composition

Abstract

The present application relates to the technical field of microbial immobilization, and discloses a preparation process of a specific carrier immobilized multifunctional microbial flora composition, comprising: mixing multifunctional microbial flora with an inducer to produce extracellular polymers, to obtain pretreated bacterial solution; adding carboxymethyl deacetyl chitin to the pretreated bacterial solution and adjusting the pH value, to form a primary wrapping suspension; immersing a porous carrier in the primary wrapping suspension, spraying a sodium phytate and calcium lactate composite aqueous solution under a pressure difference environment, and constructing a concentration gradient crosslinking network on the surface and pores of the carrier through ion bridging, wherein the present application utilizes component protonation response mechanism and nonlinear crosslinking distribution to actively open diffusion channels under a specific acid-base environment, reduce metabolic product discharge resistance, and maintain long-term metabolic activity and structural stability of the microbial flora.

Description

Technical Field

[0001] This invention relates to a process for preparing a multifunctional microbial community composition immobilized on a specific carrier, belonging to the field of microbial immobilization technology. Background Technology

[0002] Current microbial immobilization technology is used to improve the operating efficiency of bioreactor systems. By anchoring functional microbial communities to the surface of a carrier, it achieves effective enrichment of microbial biomass and enhances shock resistance. By physically modifying the carrier surface or introducing specific chemical functional groups, the adhesion strength of the interface is enhanced. This evolutionary path, which aims to pursue static connection strength, exposes deep-seated physical constraints when dealing with the complex operating conditions of large-scale continuous flow reaction systems.

[0003] In applications handling high concentrations of organic loads, the immediate release of microbial metabolites becomes a core factor limiting system performance. Because conventional immobilization layers are often densely physically encapsulated, protons, carbon dioxide, and organic acids produced by microorganisms during metabolism accumulate in the microenvironment due to diffusion resistance within the colonization layer. This leads to the buildup of metabolites within the colonization layer. The localized accumulation of these products in the interfacial microspace causes a rapid decrease in pH, far exceeding the tolerance limits of the functional microbiota, thus inducing systemic biofilm inactivation. Existing preparation processes lack physiological metabolic feedback sensing at the structural control level; for example, publication number C... Chinese invention patent N106636060A discloses a microbial immobilization carrier, microbial immobilization equipment, and microbial immobilization method. It utilizes polyvinyl alcohol and sodium alginate to construct a composite embedding system and supplements it with spiral extrusion and three-stage cross-linking processes to improve immobilization efficiency and reduce cell release rate. However, the solidified network is a static physical barrier. After the system is cross-linked and solidified, the pore size of the internal diffusion channels is locked, and the interfacial permeability cannot be adjusted according to the fluctuation of metabolic intensity. Under extreme metabolic conditions, the conflict between the high-intensity embedding mechanism and mass transfer efficiency leads to the formation of mass transfer dead zones inside the immobilized layer, and even causes biofilm detachment due to the inability to respond to metabolic signals in a timely manner.

[0004] Therefore, how to construct a dynamic immobilization system that can respond to microbial metabolic signals and generate physical conformation regulation, and achieve a deep synergy between immobilization strength and mass transfer efficiency without changing the properties of the carrier material, has become the technical problem to be solved by this invention. Summary of the Invention

[0005] To address the problems mentioned in the background art, the technical solution of the present invention is as follows: A process for preparing a multifunctional microbial community composition immobilized on a specific carrier, comprising the following steps: Step S101, induction step, the cell concentration is... to A multifunctional microbial community suspension was mixed with L-proline as an inducer, wherein the mass concentration of L-proline was 0.2% to 0.5%, and the mixture was stirred at a temperature of 37°C and a stirring speed of [missing information]. Under these conditions, the microbial community was treated for 4 hours to produce metabolites containing extracellular polymers, resulting in a pretreated bacterial solution. Step S102, coating step: add 1.5% to 2.5% by mass of carboxymethyl deacetylated chitin to the pretreated bacterial solution and adjust the pH of the system to 7.5 to 8.0. Through the charge extension characteristics of carboxymethyl deacetylated chitin in a slightly alkaline environment, it causes molecular chain entanglement with multifunctional microbial flora and metabolites to obtain a primary coating suspension. Step S103, cross-linking and curing step, to cross-link the pore size of 50 Up to 200 μm and specific surface area is to The porous carrier was immersed in the primary encapsulation suspension and kept at 0.04 MPa for 3 minutes to remove air from the porous carrier. A composite aqueous solution of sodium phytate and calcium lactate is sprayed onto the surface of a porous carrier at a flow rate of 1:3. The total mass percentage concentration of the composite aqueous solution is 2.5% to 3.0%. The multivalent anionic centers of phytate, calcium ions, and carboxyl groups in carboxymethyl deacetylated chitin form a multi-level ionic bridging network, generating a cross-linked network with a concentration gradient on the surface and within the pores of the porous carrier.

[0006] Preferably, the degree of deacetylation of the carboxymethyl deacetylated chitosan in step S102 is greater than 90%, and its degree of carboxymethyl substitution is 0.6 to 0.8; the carboxymethyl deacetylated chitosan undergoes protonation-induced volume shrinkage when the pH of the system drops to 5.5, and the volume shrinkage opens diffusion channels in the cross-linked network to reduce the resistance to the emission of metabolites in the multifunctional microbial community aggregation area; the mass percentage of carboxymethyl deacetylated chitosan in the final composition formed in the preparation process is 2.0% to 4.5%.

[0007] Preferably, the mass percentage of the multifunctional microbial community suspension is 15% to 25%, based on the total mass of materials in the preparation process being 100%.

[0008] Preferably, the porous carrier in step S103 is made of polyurethane or polystyrene, and the mass percentage of the porous carrier in the preparation process is 20% to 30%.

[0009] Preferably, in step S103, the composite aqueous solution is distributed from the outside to the inside of the pores of the porous carrier by a pressure difference generated by a pressure of 0.04 MPa, thereby generating a shear-resistant substrate with a shear strength greater than 15 Pa at the interface of the porous carrier.

[0010] Preferably, after step S103, a washing and activation step is further included: the solidified multifunctional microbial community composition is washed with sterile water, placed in a glucose solution with a mass fraction of 0.1%, and activated by standing at 25°C for 12 hours.

[0011] Preferably, in step S103, the mass of sodium phytate added satisfies the following quantitative relationship: ,in, The dosage of sodium phytate is expressed in grams. The cell concentration of the multifunctional microbial community suspension is expressed in CFU / mL. This represents the total volume of the primary encapsulated suspension, in mL. The average pore size of the porous carrier is expressed in units of 1000 ppm. ; This is the structural stability coefficient, in units of... Its value ranges from 0.12 to 0.15.

[0012] Preferably, in the multifunctional microbial community composition formed by the preparation process, the crosslinking density near the porous carrier wall is higher than the crosslinking density in the multifunctional microbial community aggregation area.

[0013] Preferably, the multifunctional microbial community consists of photosynthetic bacteria, nitrifying bacteria, and lactic acid-producing bacteria in a cell number ratio of 1:1:2.

[0014] Preferably, before the multifunctional microbial community suspension is processed in step S101, the bacterial cells are collected by centrifugation and washed three times with physiological saline with a mass fraction of 0.9%, and then resuspended in sterile water to adjust to a predetermined concentration.

[0015] Compared with the prior art, the beneficial effects of the present invention are: 1. In the multifunctional microbial community composition, the conformational contraction characteristics of the amphoteric charge regulator within a specific pH range enable dynamic regulation of the microbial metabolite emission channels, solving the technical bottleneck of microbial inactivation caused by the accumulation of metabolites inside the immobilized layer. The isoelectric point of carboxymethyl deacetylated chitin is limited to the pH window when microbial metabolism is vigorous. When microbial acid production causes the pH of the microenvironment to decrease and approach the isoelectric point, the amphoteric polymer segments transform from the extended state to the aggregated state, generating micron-level steric contraction as a whole. This instantaneously increases the diffusion channels inside the composition, accelerates the cross-interface transfer of protons and organic acid metabolites, prevents acidosis in the microbial community, and restores the encapsulated state after the metabolic load decreases and the pH rises, maintaining the long-term metabolic activity stability of the system.

[0016] 2. Metabolic inducing factors are used to induce the microbial community to actively secrete extracellular polymers, and a bio-anchoring network is constructed with amphoteric polymers and cross-linking synergists, transforming the traditional passive physical adsorption into active biochemical co-colonization. L-proline under 150 rpm stirring conditions induces microorganisms to secrete extracellular polysaccharides rich in hydrophobic groups, hydroxyl groups, and amino groups. These biomacromolecules serve as active sites, intertwining with the charge centers of carboxymethyl deacetylated chitin and the multivalent anion centers provided by sodium phytate. Through a multi-level ion bridging network mediated by phytate phosphate groups, a high-shear-strength base layer is formed at the carrier interface, achieving a structured arrangement of the microbial community and the carrier from the inside out, avoiding avalanche-like shedding under high load operation.

[0017] 3. By constructing a structured immune barrier with elastic deformation capability through the nonlinear distribution of crosslinking density at the interface between the microbial aggregation zone and the carrier, the dual constraints of fixation strength and mass transfer efficiency are balanced. Due to the use of dripping and spraying at a specific flow rate and vacuum filtration in the process, the sodium phytate and calcium lactate composite solution generates a concentration gradient in the carrier pores, making the crosslinking density near the carrier wall higher than that in the core area of ​​the microbial community. When subjected to fluid shear force, this heterogeneous structure can absorb energy through the dynamic breaking and recombination of ionic bonds, generating a nonlinear mechanical response to buffer hydraulic load fluctuations, ensuring that the composition can maintain the complete biofilm morphology and its uninterrupted metabolic capacity even under extreme conditions. Attached Figure Description

[0018] Figure 1 This is a flowchart illustrating the induced coating and gradient crosslinking preparation process of the multifunctional microbial community composition of the present invention. Figure 2 This is a schematic diagram of the raw material components, cell ratio, and structural composition of the multifunctional microbial community composition of the present invention. Detailed Implementation

[0019] The following disclosure is for illustrative purposes only and is not intended to limit the scope of protection of the present invention.

[0020] This invention provides a process for preparing a multifunctional microbial community composition immobilized on a specific carrier, which solves the technical problem of microbial community inactivation caused by the accumulation of metabolites during microbial immobilization. In the induction phase, the cell concentration is... to Multifunctional microbial community suspension and as an inducer -Proline mixture; multifunctional microbial community composed of photosynthetic bacteria, nitrifying bacteria and lactic acid-producing bacteria The ratio of cell number to composition, The mass concentration of -proline is to The mixed system at a temperature of And the stirring speed is Under the conditions of processing The process induces a multifunctional microbial community to produce metabolites containing extracellular polymers, resulting in a pretreated bacterial solution. During the coating stage, a pretreated bacterial solution is added with a mass percentage concentration of [missing information]. to Carboxymethyl deacetylated chitosan and the system were adjusted. Value to The degree of deacetylation of carboxymethyl deacetylated chitosan is greater than that of carboxymethyl deacetylated chitosan. Its degree of carboxymethyl substitution is to In a slightly alkaline environment, carboxymethyl deacetylated chitin undergoes molecular chain entanglement, encapsulating multifunctional microbial flora and metabolites to obtain a primary encapsulated suspension; the mass percentage of carboxymethyl deacetylated chitin in the composition is [missing information]. to When the system Value dropped to At this time, carboxymethyl deacetylated chitin undergoes protonation-induced volume shrinkage, opening diffusion channels in the cross-linked network and reducing the resistance to metabolite excretion.

[0021] During the cross-linking and curing stage, the pore size is to And the specific surface area is to A porous carrier is immersed in a primary encapsulation suspension; the porous carrier is made of polyurethane or polystyrene, and its mass percentage in the composition is [missing information]. to ;exist Maintain in the environment To remove air from the porous carrier; A composite aqueous solution of sodium phytate and calcium lactate is sprayed onto the surface of a porous carrier at a flow rate of [value missing]; the mass ratio of sodium phytate to calcium lactate is [value missing]. The total mass percentage concentration of the composite aqueous solution is to The multivalent anionic centers of phytate form a multi-level ionic bridging network with calcium ions and the carboxyl groups in carboxymethyl deacetylated chitin, generating a cross-linked network with a concentration gradient on the surface and within the pores of the porous carrier. The cross-linking density near the porous carrier wall is higher than that in the multifunctional microbial community aggregation zone. The composite aqueous solution passes through... The pressure difference generated by the pressure decreases from the outside to the inside within the pores of the porous carrier, and a shear strength greater than 1 is generated at the interface of the porous carrier. The shear-resistant sublayer; the cross-linking depth is controlled by adjusting the pressure value of the negative pressure environment and the flow rate of the composite aqueous solution. The pressure gradient generated by the negative pressure environment drives the composite aqueous solution to penetrate deep into the pores of the porous carrier. The ion bridging reaction rate between phytate and carboxymethyl deacetylated chitin determines the concentration distribution of the solute along the penetration path. When the flow rate is set to... And the pressure is set to At that time, by utilizing the competitive mechanism of solute penetration and rapid cross-linking reaction, near the wall of the porous support... The crosslinking density is high within the range to form a shear-resistant substrate, while the crosslinking density is higher at a distance from the wall. The multifunctional microbial community aggregation zone has a cross-linking density reduced to that at the wall surface. to Thus, greater than 10 ... The interfacial shear strength and the flexible coating that preserves the conformational response space.

[0022] The width of the internal diffusion channels of the composition and the control of the concentration gradient were verified by a tracer probe permeation experiment coupled with the adjustment of negative pressure impregnation parameters. The composition was immersed in water. for and Standard buffer solution, add molecular weight to the system Sodium fluorescein was used as a tracer molecule. The diffusion rate of the tracer molecule within the cross-linked network was measured using laser confocal microscopy. The diffusion channel width under different acidic and alkaline conditions was calculated using the equivalent medium diffusion equation. The degree of volume shrinkage induced by carboxymethyl deacetylation of the chitin was characterized. Step S103 involved controlling… Negative pressure environment maintenance time and The spray flow rate utilizes the tortuous flow resistance within the carrier pores to generate a nonlinear decay at the solute wetting front, causing a decreasing ion bridging reaction in the sodium phytate and calcium lactate composite aqueous solution along the permeation path. This constructs a high-density shear-resistant sublayer near the carrier wall and a low-density cross-linked layer with proton-responsive characteristics in the bacterial aggregation zone; the dosage of sodium phytate... The following quantitative relationship must be satisfied: ,in, The dosage of sodium phytate is expressed in units of [unit missing]. ; This is the structural stability coefficient, in units of... Its value range is to ; The cell concentration of the multifunctional microbial community suspension is expressed in units of... ; The total volume of the primary encapsulated suspension, in units of ; The average pore size of the porous carrier is expressed in units of 1000 ppm. Calibrate the stability coefficient of porous carrier structure The parameters were determined using a step variable test method. for to Within the interval A series of predictive sample groups were prepared using a step gradient method. The biofilm loss rate of the sample groups under shear force fluctuations was measured using a hydrodynamic testing unit. The distribution density of the cross-linked network at the carrier interface was observed using scanning electron microscopy, and the interfacial shear strength was measured to reach [value missing]. And the average thickness of the shear-resistant substrate is stable at to When the sample group values ​​are used as the structural stability coefficient of the production batch, the values ​​of the sample group are used as the structural stability coefficient. Establish the quality of sodium phytate dosing With the average pore size of the carrier and cell concentration The correspondence relationship ensures that the cross-linked network maintains interfacial stability and reserves space for conformational response of molecular chain segments; after curing, the cured multifunctional microbial community composition is washed with sterile water; the composition is then placed in a container with a mass fraction of In glucose solution, Let stand at ℃ activation.

[0023] Example 1: When the influent chemical oxygen demand concentration is In industrial dairy processing wastewater treatment environments, microorganisms produce lactic acid during the degradation of organic matter, leading to localized degradation within the carrier. The value fell to The following; nitrifying bacteria are inhibited by the accumulation of metabolic products, and due to fluid shear forces... Fluctuations caused the biofilm on the carrier surface to peel off. Using the preparation process described in the specific embodiments above, a cell concentration of [missing information] was taken. A multifunctional microbial community suspension; the multifunctional microbial community consists of photosynthetic bacteria, nitrifying bacteria, and lactic acid-producing bacteria, arranged in... The cell number ratio composition; the suspension was mixed with a mass concentration of of -proline mixture, in ℃ and stirring speed is Under the conditions of processing A pretreated bacterial solution was obtained; a bacterial solution with a mass percentage concentration of [missing information] was added to the pretreated bacterial solution. Carboxymethyl deacetylated chitosan and the system were adjusted. Value to The degree of deacetylation of carboxymethyl deacetylated chitosan is [value missing]. Its degree of carboxymethyl substitution is Utilizing the electrostatic properties of molecular chains under alkaline conditions, a primary encapsulated suspension is obtained by coating a multifunctional microbial community; the average pore size is selected. for Specific surface area is The polyurethane porous carrier is set as a mass percentage in the composition. According to the formula Set the structural stability coefficient for ; in the primary encapsulation of the total volume of the suspension for And cell concentration for Under the given conditions, the dosage of sodium phytate was calculated. for ,in, The dosage of sodium phytate is expressed in units of [unit missing]. ; This is the structural stability coefficient, in units of... ; The cell concentration of the multifunctional microbial community suspension is expressed in units of... ; The total volume of the primary encapsulated suspension, in units of ; The average pore size of the porous carrier is expressed in units of 1000 ppm. .

[0024] The porous carrier was immersed in the primary encapsulation suspension. Maintaining negative pressure environment ;by A composite aqueous solution of sodium phytate and calcium lactate is sprayed onto the surface of a porous carrier at a flow rate of [value missing]; the mass ratio of sodium phytate to calcium lactate is [value missing]. The total mass percentage concentration of the composite aqueous solution is During the spraying process, the multivalent anionic centers of phytate ions form a multi-level ionic bridging network with calcium ions and the carboxyl groups in carboxymethyl deacetylated chitin; the pressure difference causes the crosslinking agent to be distributed in a gradient from the outside to the inside, generating a shear strength greater than that at the interface of the porous carrier. After the shear-resistant substrate is prepared, the composition is washed with sterile water and placed in a container with a mass fraction of [missing information]. In glucose solution, Let stand at ℃ Activation; during continuous operation, when microbial metabolism produces acid, causing local... Value dropped to At this time, carboxymethyl deacetylated chitin undergoes volume shrinkage due to protonation, opening diffusion channels in the cross-linked network and reducing the migration resistance of lactate ions; the system's total nitrogen removal rate remains at The above results were observed, and the biofilm on the porous carrier surface did not peel off.

[0025] Example 2: To verify the effectiveness of the dynamic adjustment of mass transfer resistance within the microbial immobilization layer with increasing metabolic intensity, this experiment employed a continuous flow bioreactor system, including a measurement range of [missing information]. to And the accuracy is of Monitoring unit, sampling frequency is And the resolution is The fluid dynamics control unit; the experimental data were taken from a simulated dairy processing wastewater treatment process, in which the initial chemical oxygen demand (COD) concentration was adjusted by adding lactose and casein to deionized water. To evaluate the stability of the composition under real-world conditions, the frequency of introduction into the test fluid was [missing information]. And the amplitude is The periodic shear force fluctuations were simulated to monitor fluid disturbances caused by industrial pump switching, while the diffusion channel width and total nitrogen removal efficiency of different combinations were monitored as the microenvironment pH decreased. In the process parameter settings, the value of the structural stability coefficient k determined the trade-off between the mechanical strength and conformational response sensitivity of the ion-bridged network; if k is lower than... Insufficient cross-linking density between phytate and calcium ions leads to a decrease in the interfacial shear strength of the underlying layer, under shear stress of [missing value]. The biofilm loss rate increases; if k is higher than... Excessive cross-linking constraints limit the shrinkage space of the carboxymethyl deacetylated chitosan molecular chain during protonation, hindering the opening of diffusion channels. The experimental setup included the sample group of this invention, the control sample group, and the sample group outside the range, as shown in Table 1.

[0026] Table 1: Actual Performance Data of Different Groups Analyzing the measurement data in Table 1, the sample group of the present invention... Value dropped to At the inflection point, carboxymethyl deacetylated chitin undergoes protonation-induced conformational contraction, and the diffusion channel width changes from the normal state. Increase to This allows for the immediate efflux of metabolic products; control group A lacks amphoteric regulatory components, maintaining mass transfer channels at extremely low levels, internal acid accumulation inhibits nitrifying bacteria activity, and the total nitrogen removal rate decreases to [missing value]. Although control group B showed responsiveness, its interfacial shear strength was only [missing information] due to the lack of gradient cross-linking structure. The bacterial residue rate decreased; in the continuous In the gradient pressure test, as the organic load of the wastewater increased from low to high, the internal microenvironment of the sample group of this invention... Value presentation by Towards The downward trend, while the diffusion rate of metabolites monitored by the system and The degree of decrease in the value showed a positively correlated nonlinear enhancement characteristic; the physical conformation regulation mechanism triggered by this metabolic signal ensured that the bacterial community maintained a stable metabolic flux under extreme load shocks; when the experimental group carrier was observed with a scanning electron microscope, the ion bridging network formed on the carrier wall was dense and continuous, while micron-sized gaps were retained near the center of the bacterial community.

[0027] Example 3: This example combines Figures 1 to 2 The preparation process of a multifunctional microbial community composition immobilized on a specific carrier is described, such as... Figure 1 As shown, step S101 is the induction step, in which the multifunctional microbial community suspension is mixed with the inducer L-proline and treated at 37°C with stirring to induce the community to produce metabolites containing extracellular polymers to obtain a pretreated bacterial solution. Step S102 is the coating step, in which carboxymethyl deacetylated chitin is added to the pretreated bacterial solution and the pH is adjusted to 7.5 to 8.0. Molecular chain entanglement occurs by utilizing the charge extension characteristics under alkaline conditions to obtain a primary coated suspension. Finally, step S103 is the cross-linking and curing step, in which the porous carrier is immersed in the primary coated suspension and the air is degassed at 0.04 MPa. A composite aqueous solution of sodium phytate and calcium lactate is sprayed on the carrier to generate a cross-linked network with a concentration gradient on the carrier surface and in the pores by using ion bridging.

[0028] like Figure 2 As shown, the core of this system includes a porous carrier made of polyurethane or polystyrene with a pore size of 50 μm to 200 μm, and an extracellular polymer EPS induced by L-proline. The multifunctional microbial community consists of photosynthetic bacteria, nitrifying bacteria, and lactic acid-producing bacteria in a cell ratio of 1:1:2. It is externally coated with carboxymethyl deacetylated chitin with a degree of deacetylation greater than 90% and a mass percentage of 2.0% to 4.5%. The overall structure is fixed by a sodium phytate-calcium ion crosslinking network with a sodium phytate to calcium lactate ratio of 1:3 and the network exhibits a concentration gradient distribution. The final composition contains the porous structure shown in the figure and microorganisms A and B distributed within it.

[0029] Example 4: In a high-concentration organic acid wastewater treatment scenario, the microbial community produces a large amount of acetic acid during the degradation of organic matter, leading to an increase in the local proton concentration inside the immobilized layer, thus affecting the microenvironment. The value starts from the initial Descending to The threshold; due to the static hindrance effect of conventional immobilized structures, metabolites cannot migrate outward across the interface in a timely manner, thus forming a mass transfer dead zone that inhibits biological activity in the bacterial community aggregation area. To address this mass transfer dead zone problem, this embodiment utilizes the isoelectric point characteristics of carboxymethyl deacetylated chitin, when the local microenvironment... Value dropped to At this time, the carboxyl groups of the side chains of carboxymethyl deacetylated chitin molecules are protonated, and the electrostatic repulsion between molecules is transformed into a contractile force driven by hydrogen bonds. This causes carboxymethyl deacetylated chitin to transform from an extended random coil conformation to an aggregated granular conformation, thereby generating micron-sized steric gaps between the physical lattice points of the cross-linked network, providing diffusion channels for the cross-interface migration of organic acid anions.

[0030] To ensure the decoupling of the aforementioned response mechanism from mechanical strength, this preparation process employs a negative pressure assisted spraying method, by setting... The negative pressure environment creates a pressure gradient pointing towards the wall within the pores of the porous carrier. A sodium phytate and calcium lactate composite aqueous solution was introduced at a flow rate of [value missing]. The polyvalent anionic centers of phytate ions react with calcium ions and the carboxyl groups in carboxymethyl deacetylated chitin to form an ion-bridging reaction. Because the solute permeation rate is limited by the pore tortuosity, a high-density shear-resistant sublayer preferentially forms near the porous carrier wall, while a responsive layer with lower cross-linking density and conformational adjustment space forms near the core of the bacterial community. This results in the construction of a shear strength greater than [value missing]. The asymmetric network, the dosage of sodium phytate Follow the following quantitative relationship: ,in, The dosage of sodium phytate is expressed in units of [unit missing]. ; The structural stability coefficient is... to The range is selected to balance the interface anchoring strength and the chain segment shrinkage rate; The cell concentration of the multifunctional microbial community suspension is expressed in units of... ; The total volume of the primary encapsulated suspension, in units of ; The average pore size of the porous carrier is expressed in units of 1000 ppm. Under these implementation conditions, experimental measurement data show that when the internal microenvironment... Value dropped to At that time, the width of the diffusion channels inside the multifunctional microbial community composition increased from the normal value. Expansion to The corresponding metabolite migration rate across the interface increases to the initial state. Times, while the fluid shear force is Under operating conditions, the bacterial residue rate remained at .

[0031] Example 5: Before different batches of porous carriers and multifunctional microbial communities were put into production, the average pore size of the porous carriers was obtained. and the cell concentration of multifunctional microbial community suspension And substitute the collected values ​​into the quantization relationship Calculate the mass of sodium phytate added. ;in, The dosage of sodium phytate is expressed in units of [unit missing]. ; This is the structural stability coefficient, in units of... ; Cell concentration, in units of ; The total volume of the primary encapsulated suspension, in units of ; The average pore size of the porous carrier is expressed in units of 1000 ppm. ;exist to Adjust the shear force within a certain range and measure the biofilm loss rate. When the biofilm loss rate is below a certain value... Furthermore, the multifunctional microbial community composition is in Value Under the condition of conformational contraction response, the structural stability coefficient of the current production batch is fixed. The value.

[0032] Before treating wastewater with different background conductivity, by having Determination of the critical conformational transition of carboxymethyl deacetylated chitin using a unit-interval pH gradient solution Point; if critical Point relative to The absolute value of the deviation is greater than Then, adjust the spraying time of the sodium phytate and calcium lactate composite aqueous solution in step three. Utilizing the limiting effect of phytate on the carboxyl protonation process, the trigger interval for opening the diffusion channel is corrected to... Value The range allows the final product to achieve a balance between structural stability and metabolic efflux flux in water environments with different ionic strengths.

[0033] Example 6: In large-scale production scenarios involving multiple batches of material flow, to address the technical problem of uneven primary coating thickness caused by differences in the solubility of carboxymethyl deacetylated chitosan, a pre-matching procedure was implemented between the density of the active component of the raw material and the dynamic viscosity of the material. Before starting the preparation process, carboxymethyl deacetylated chitosan was added to sterile water and stirred at a stirring speed of [missing information]. The paddle mixer is used for mixing, utilizing a measurement accuracy of The rotational viscometer was used to determine the dynamic viscosity of the primary encapsulated suspension. Based on measured values, the volumetric addition rate of the pretreated bacterial solution is adjusted using a precision metering pump to ensure that the coating thickness formed on the porous carrier surface and within the pores of the material entering the subsequent negative pressure impregnation step is within acceptable limits. to Within a certain range, the initial mass transfer efficiency is maintained by compensating for deviations in the physical properties of the raw materials.

[0034] When the prepared multifunctional microbial community composition is transferred to a mass fraction of During activation treatment in glucose solution, a metabolic recovery procedure based on a stepwise increase in dissolved oxygen concentration was implemented to regulate the physical balance between the oxygen demand of the microbial community and the mass transfer limitation of the immobilized layer; a measurement range of [missing information] was deployed in the activation container. to And the response time is less than Dissolved oxygen electrode for real-time monitoring of dissolved oxygen concentration in the system. The air supply rate of the ventilation device is adjusted by the control unit to make... From the initial Beginning gradient per interval Increasing the load, this gradual regulation of oxygen partial pressure from low to high guides microorganisms to complete the synthesis and immobilization of intracellular enzyme systems through physical anchoring at the interface. At the end of the activation cycle, the biological activity is confirmed to have recovered to the preset index by measuring the unit degradation rate of organic nutrients in the solution, so that the product can complete the transformation from the dormant state to the stable metabolic state before facing high shear conditions.

[0035] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention.

[0036] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A process for preparing a multifunctional microbial community composition immobilized on a specific carrier, characterized in that, Includes the following steps: Step S101, induction step, the cell concentration is... to A multifunctional microbial community suspension was mixed with L-proline as an inducer, wherein the mass concentration of L-proline was 0.2% to 0.5%, and the mixture was stirred at a temperature of 37°C and a stirring speed of [missing information]. Under these conditions, the microbial community was treated for 4 hours to produce metabolites containing extracellular polymers, resulting in a pretreated bacterial solution. Step S102, coating step: add 1.5% to 2.5% by mass of carboxymethyl deacetylated chitin to the pretreated bacterial solution and adjust the pH of the system to 7.5 to 8.

0. Through the charge extension characteristics of carboxymethyl deacetylated chitin in a slightly alkaline environment, it causes molecular chain entanglement with multifunctional microbial flora and metabolites to obtain a primary coating suspension. Step S103, crosslinking and curing step, to crosslink and cure materials with pore sizes of 50 μm to 200 μm and specific surface areas of... to The porous carrier was immersed in the primary encapsulation suspension and kept at 0.04 MPa for 3 minutes to remove air from the porous carrier. A composite aqueous solution of sodium phytate and calcium lactate is sprayed onto the surface of a porous carrier at a flow rate of 1:

3. The total mass percentage concentration of the composite aqueous solution is 2.5% to 3.0%. The multivalent anionic centers of phytate, calcium ions, and carboxyl groups in carboxymethyl deacetylated chitin form a multi-level ionic bridging network, generating a cross-linked network with a concentration gradient on the surface and within the pores of the porous carrier.

2. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, The degree of deacetylation of carboxymethyl deacetylated chitin in step S102 is greater than 90%, and its degree of carboxymethyl substitution is 0.6 to 0.

8. When the pH of the system drops to 5.5, carboxymethyl deacetylated chitin undergoes protonation-induced volume shrinkage, which opens diffusion channels in the cross-linked network to reduce the resistance to the emission of metabolites from the multifunctional microbial community aggregation area. The mass percentage of carboxymethyl deacetylated chitin in the final composition formed in the preparation process is 2.0% to 4.5%.

3. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, Based on the total material mass in the preparation process being 100%, the mass percentage of the multifunctional microbial community suspension is 15% to 25%.

4. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, The porous carrier in step S103 is made of polyurethane or polystyrene, and the mass percentage of the porous carrier in the preparation process is 20% to 30%.

5. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, In step S103, the composite aqueous solution is distributed from the outside to the inside of the porous carrier through a pressure difference generated by a pressure of 0.04 MPa, thereby generating a shear-resistant sublayer with a shear strength greater than 15 Pa at the interface of the porous carrier.

6. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, After step S103, a washing and activation step is also included: the solidified multifunctional microbial community composition is washed with sterile water, placed in a 0.1% glucose solution, and activated by standing at 25°C for 12 hours.

7. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, In step S103, the added mass of sodium phytate satisfies the following quantitative relationship: ,in, The dosage of sodium phytate is expressed in grams. The cell concentration of the multifunctional microbial community suspension is expressed in CFU / mL. This represents the total volume of the primary encapsulated suspension, in mL. The average pore size of the porous carrier is expressed in units of 1000 ppm. ; This is the structural stability coefficient, in units of... Its value ranges from 0.12 to 0.

15.

8. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, In the multifunctional microbial community composition formed by the preparation process, the crosslinking density near the porous carrier wall is higher than the crosslinking density in the multifunctional microbial community aggregation area.

9. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, The multifunctional microbial community is composed of photosynthetic bacteria, nitrifying bacteria, and lactic acid-producing bacteria in a cell number ratio of 1:1:

2.

10. The preparation process of a multifunctional microbial community composition immobilized on a specific carrier according to claim 1, characterized in that, Before the multifunctional microbial community suspension is processed in step S101, the bacterial cells are collected by centrifugation and washed three times with physiological saline with a mass fraction of 0.9%, and then resuspended in sterile water to adjust to the predetermined concentration.

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