Bioreactor system with metabolic flow analysis as the core
The biological system and its environment in a bioreactor form a relatively closed ecosystem. In this ecosystem, there are interactions between biological systems and environmental factors. The phenotype of biological systems, especially those dominated by cells, is closely related not only to genotype, but also to the micro or macro environmental conditions in which the cells are located (such as nutrient types, pH, temperature, dissolved oxygen, mixing and transfer characteristics of bioreactors, etc.).
In other words, genotype and environment jointly determine the phenotypic characteristics of biological systems. On the other hand, biological processes have highly nonlinear and time-varying characteristics, making it difficult to characterize their complex ontological features using macroscopic dynamics aimed at detecting and controlling environmental operating parameters.
Therefore, it is necessary to systematically analyze the metabolic changes of cells based on the metabolic characteristics related to parameters during the fermentation process, emphasizing that the physiological state of cells is related to parameters and is the result of the transfer, conversion, and balance of materials, energy, or information in bioreactors.
Although its micro influencing factors may only occur at a certain scale at the level of genes, enzymes, cells, or reactors, they will ultimately be reflected in macro processes, providing clues for studying data correlation analysis methods at different scales in bioreactors.
An ideal bioreactor system should detect and analyze operating parameters and state parameters at different scales as much as possible, so as to potentially construct an optimized external environment that maximizes the gene expression and metabolic regulation of microorganisms for the biosynthesis of a certain target product and maximizes the accumulation of the target product.
The above indicates the engineering science issues of biological processes, which have evolved from macroscopic dynamics research to multi-scale theoretical methods based on biological process information processing, in order to guide the development of engineering technologies related to biological processes with bioreactors as the core.
Bailun adopts a metabolic flow multi-scale biological reaction system, which includes various advanced sensors for the detection of cell physiological metabolic characteristics, sensor reactors and control systems for micro metabolic flow analysis of biological processes, computer software packages for the correlation analysis of cell physiological metabolic parameters, and computer Internet systems for data processing and remote analysis of fermentation processes.
This system is consistent with the highest configuration in Europe, reducing the interference of human factors on scientific research, improving data repeatability and accuracy, reducing labor and material costs, without the need for dedicated personnel on duty, and can achieve computer remote control and wireless monitoring.
The can lid automatic opening system, and all parameters can be automatically controlled. Being able to obtain as much biological information as possible at various scales of the biological processing process, and then based on the principle of multi-scale parameter correlation, through real-time data processing of computer software, identifying key parameters for process optimization based on parameter correlation characteristics in massive data, and using them to guide process operations, equipment design, or strain screening and transformation, ultimately achieving process optimization and scaling up.
The system has been successfully applied to optimize the production process of various products, greatly improving the capacity of fermentation units. Its optimized process can generally be directly scaled up from tens of liters of fermentation tanks to industrial production fermentation tanks of hundreds of cubic meters.
The bioreactor system with metabolic flow analysis as its core is equipped with advanced sensing systems. In addition to conventional detection and control parameters such as pH, temperature, stirring speed, dissolved oxygen, and rotor flow meter, a thermal mass flow meter (for precise measurement and control of intake flow rate to ensure it is not affected by intake pressure), a fermentation broth weighing system, exhaust gas oxygen and CO2 measuring instruments (exhaust gas composition analyzer or process exhaust gas mass spectrometer), and a top mounted silicone oil pressure sensor are also configured to accurately measure the oxygen consumption rate (OUR) and CO2 release rate (CER) of biological processes.
Micro metabolic flow analysis sensor reactors and control systems, in situ live cell concentration online measuring instruments, cell morphology online microscopic observation instruments, and other online sensors can also be configured as needed. Based on the direct parameters obtained from the measurement, important physiological and metabolic state parameters such as CER, OUR, and RQ can be calculated using computer software packages for cell physiological and metabolic parameter analysis.
The process exhaust gas mass spectrometer is mainly used for measuring the concentrations of O2, CO2, and N2 in exhaust gas. This mass spectrometer uses an electron bombardment ion source. After online pretreatment, the exhaust gases from different fermentation tanks are continuously input into the ionization chamber through a multi-channel rotary valve to form ions.
Using the motion law of charged particles in the electric field, the quadrupole mass analyzer separates the ions generated by the ion source according to their mass to charge ratio (mass to charge ratio, m/z), measures the distribution of ion mass intensity, and obtains information on compound types and concentrations, accurately reflecting changes in the composition of fermentation exhaust gases.
The measurement results can be input into a specialized software package designed for the fermentation process, enabling correlation analysis with other fermentation parameters. The exhaust gas mass spectrometer can measure volatile gas components with a relative molecular weight of up to 300, so it can also detect small molecule substances such as ethanol and methanol according to fermentation needs.
Biomass is an important parameter in the fermentation process. At present, it is usually obtained through offline measurements using classic methods such as dry mass and turbidity. The in-situ live cell concentration online analyzer can not only perform real-time online measurement, but also obtain more biologically meaningful live cell concentrations.
This is particularly suitable for fermentation processes that contain insoluble solid substances in the culture medium. The principle of the live cell analyzer is based on 0 In an alternating electric field with a frequency range of 1-10MHz, non-conductive polarization occurs on the cell surface such as the cell membrane in the fermentation broth, making living cells with intact cytoplasmic membranes essentially like capacitors (the non-conductive nature of lipid cytoplasmic membranes generally leads to charge growth), while dead cells, lysed cells, cell fragments, bubbles, and other matrix components are essentially non polarizable.
By applying an alternating electric field within the frequency range using dual electrodes, the measured capacitance between the electrodes depends on the cell type and size, and is proportional to the concentration of viable bacteria within a certain range.
The in situ live cell concentration online analyzer is applicable to various animal and plant cells, yeast, bacteria, algae, etc. But it is not suitable for fermentation processes that require acid-base neutralizing agents as the target product, as excessive ion strength in the fermentation broth can interfere with the accurate measurement of capacitance.
