DD226902A1 - METHOD FOR THE TARGETED INFLUENCE OF THE DRIVING OF MICROBIAL PROCEDURES - Google Patents

Abstract

The invention relates to a method for the preparation of Fuehrungsgroessen for the operation of process stages for the production and processing of microorganisms. The aim of the invention is the targeted influencing of the technical process with respect to the production of products with certain desired properties, eg. B. Protein content, extractability. According to the invention, this is achieved by carrying out an acid-base titration and calculating the desired sizes from the titration curve, which-if the relationships to the target quantities are known-themselves become the lead quantity or enter into it.

Description

-2- 670

embodiment

After correcting the primary measurement data (pH value, consumption of titration solution), the number of η of the existing reaction partners is calculated from the sequence of measurements which corresponds to the product-specific titration solution consumption at a given pK value.

Table 1 Dependence of the Number of Reactants Available on the pH of the Fermenter pH in the fermenter 4.1 4.3 4.4 4.7

η in 10 '° H ₃ O + / cell 3.1 2.2 1.7 1.4

As Table 1 shows, there is a dependency between fermentation pH and the number of reactants found, whose pK = 2.5, on the cell surface or in near-surface areas. In a first approximation, therefore, is the production of microorganisms with defined surface properties, hier. B. achievable by pH control.

Table 2 shows the qualitative changes observed during major procedures:

Table 2 pK value before and after denaturation in the pH range from 2.2 to 4.0 natively pK = 2.7 ± 0.1 3.3 + 0.1

denatured pK 2.3 + 0.1 2.7 + 0.1 3.3 ± 0.1

Again, the calculation of the number of reaction partners in the pK values allows an assessment and further control of the procedure.

Claims (6)

1. A method for selectively influencing the procedure microbial process stages or the processing of microorganisms, such as the production of microbial feed, and for the characterization of microbial products, preferably the surface of microorganisms, characterized in that by recording and evaluation of the acid-base titration curve of a a qualitative and quantitative characterization of the sample is carried out and that these characterization variables, because of their relationship to the target parameters of the process, are used as reference variables in sub-stages or sub-stages with a comparable task or the overall process. -1- 670 Invention claims: 2. The method according to claim 1, characterized in that the characterization by the pK values, d. H. through the turning points in the titration curve. 3. The method according to claim 1 and 2, characterized in that the characterization is carried out by the quantitative evaluation of the titration curve. 4. The method according to claim 1 to 3, characterized in that the characterization is carried out by titration between predetermined pH values. 5. The method according to claim 1, characterized in that the outward and Rücktitrationskurve or from both to be constructed hysteresis curve is used to obtain the reference variable. 6. The method according to claim 1, characterized in that the titration takes place in the form of a single addition of a defined amount of titrant. Field of application of the invention The invention relates to a method for selectively influencing the mode of operation of microbial process stages for the production and processing of microorganisms up to the end product on the basis of the characterization of the respective state of the microorganisms or of the product. Characteristic of the known technical solutions The known methods for optimizing microbial process stages are based on the detection of quickly determinable environmental factors, eg. B. pH, pO ₂ value, concentration of the nutrient additives and excipients, the exhaust gas analysis and the calculated from this material conversion. From this information, the dosage of the corresponding streams is derived (Rotenburger Symposium 1978: fermentation technology, B. Braun, Melsungen AG, lecture L. Nyeste et al., Page 141, DD-10 006, DD-159 011, SU-791 771). Further possibilities which are used for the characterization of the state of microorganisms are the measurement of the electrophoretic mobility (DE-OS-1 642 821, DD-160 012, GB-2 054 839, SU-911 294, Huber and Werner in vt "Verfahrenstechnik" 17 [1983] 1: 19-22), the fluorescence and luminescence analysis after adsorption of suitable substances on the cell surface (GB-2 116 712; GB-2 095 830; SU-951 112), the spectrophotometric examination of Cell components, eg cytochrome (DD-154 449), microorganism detection via the bioluminescence of adenosine triphosphate after cell destruction (GB-2 116 709), protein portion (DE-AS-2 365 79 6). In addition to the already mentioned investigation possibility by means of light microscope, supplemented by specific staining reactions, the various electron microscopic examination methods are still known, which currently represent the most fertile source of information about the state of a microorganism, but are also very expensive. Among the known methods of investigation of structural units of microorganisms include the statements about the cell membrane after adsorption and fluorescence (SU-1 010 126). Also known are the titration of cells without further evaluation and use of the results (DD-137149), supplemented by spectrophotometric devices / investigations (SU-924 566, SU-951 112). Furthermore, the relationships between the titration curve of a yeast culture and the physiological state of the yeast sample are known. Since ammonia solution, which simultaneously has substrate function, is used as the titration agent, these statements are complex in nature. Process control on this basis is not performed (C.A., cit., 100/1 / 4767m after Gidroliz, Lesokhim, Prom. 1983 (6) 5-7). Object of the invention The aim of the invention is a method which allows the determination of characteristic quantities, preferably surface properties, of microorganisms or the products resulting therefrom in the course of the processing and the use of these characterization variables as a reference variable for the process control after correlation with product target parameters. Object of the invention The invention has for its object to develop a method for the determination of characterization parameters, which detects essential properties of the examination object. Features of the invention According to the invention this is achieved in that the sample containing the object to be examined, preferably microorganisms, such as yeast, subjected to a separation operation and, optionally after an object-dependent purification, suspended in CO ₂ -free water in standardized proportions and subsequently subjected to acid-base titration becomes. The obtained titration curve provides the desired characterization quantities, such as the pK values of the titrated functional groups, the number of reactants present per group and further quantities obtainable from the mathematical treatment / division / integration of the titration curve (s), eg. B. areas of the hysteresis curve. Conveniently, the characterization can be carried out by a quantitative evaluation of the titration curve or by a titration between fixed pH values. Titration in the form of a single addition of a defined amount of titration agent is also possible. These characterization variables, in conjunction with the target variables of the partial or overall process, obtain the property of reference variables / target variables for controlling the technological process at a qualitatively higher level.