CN111474134A - Method for controlling butyric acid fermentation by using online near infrared - Google Patents
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- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 title claims abstract description 92
- 238000000855 fermentation Methods 0.000 title claims abstract description 85
- 230000004151 fermentation Effects 0.000 title claims abstract description 72
- 238000000034 method Methods 0.000 title claims abstract description 55
- 238000001514 detection method Methods 0.000 claims abstract description 39
- 238000004519 manufacturing process Methods 0.000 claims abstract description 13
- 238000005259 measurement Methods 0.000 claims abstract description 7
- 238000002329 infrared spectrum Methods 0.000 claims description 18
- 241000193171 Clostridium butyricum Species 0.000 claims description 17
- 238000002790 cross-validation Methods 0.000 claims description 15
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 13
- 239000008103 glucose Substances 0.000 claims description 13
- 239000002028 Biomass Substances 0.000 claims description 9
- 230000003595 spectral effect Effects 0.000 claims description 9
- 230000000813 microbial effect Effects 0.000 claims description 8
- 238000012937 correction Methods 0.000 claims description 5
- 238000009499 grossing Methods 0.000 claims description 5
- 230000002503 metabolic effect Effects 0.000 claims description 5
- 238000010606 normalization Methods 0.000 claims description 5
- 238000012847 principal component analysis method Methods 0.000 claims description 5
- 238000012569 chemometric method Methods 0.000 claims description 4
- 238000010561 standard procedure Methods 0.000 claims description 4
- 244000005700 microbiome Species 0.000 claims description 3
- 238000011156 evaluation Methods 0.000 claims description 2
- 229910000530 Gallium indium arsenide Inorganic materials 0.000 claims 1
- KXNLCSXBJCPWGL-UHFFFAOYSA-N [Ga].[As].[In] Chemical compound [Ga].[As].[In] KXNLCSXBJCPWGL-UHFFFAOYSA-N 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 11
- 238000012544 monitoring process Methods 0.000 abstract description 6
- 238000010986 on-line near-infrared spectroscopy Methods 0.000 abstract description 5
- 238000004497 NIR spectroscopy Methods 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 239000000758 substrate Substances 0.000 abstract description 3
- 238000010923 batch production Methods 0.000 abstract description 2
- 230000000694 effects Effects 0.000 abstract description 2
- 230000009123 feedback regulation Effects 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 238000001228 spectrum Methods 0.000 abstract description 2
- 230000003287 optical effect Effects 0.000 abstract 1
- 238000003908 quality control method Methods 0.000 description 7
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 239000000126 substance Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 4
- JBRZTFJDHDCESZ-UHFFFAOYSA-N AsGa Chemical compound [As]#[Ga] JBRZTFJDHDCESZ-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 229910052738 indium Inorganic materials 0.000 description 3
- APFVFJFRJDLVQX-UHFFFAOYSA-N indium atom Chemical compound [In] APFVFJFRJDLVQX-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000006052 feed supplement Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3577—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing liquids, e.g. polluted water
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/359—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using near infrared light
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Abstract
The invention provides a method for controlling butyric acid fermentation by using online near infrared, and relates to the technical field of near infrared spectroscopy. Through the online detection of data and the combination of actual production process conditions, the production activity is subjected to feedback regulation, so that the conversion rate of the substrate and the content of a target product are improved, the consumption of raw materials is reduced, and the economic benefit of an enterprise is increased. The invention utilizes the on-line near infrared spectrum technology to automatically monitor, count and feed back parameters of the butyric acid anaerobic fermentation process in real time, has the advantages of high detection speed, nondestructive measurement, no pollution by optical detection, no foreign matter introduction, no influence on the fermentation process, simultaneous measurement of a plurality of physicochemical parameters and the like, and the detected data can be fed back to a control system without delay, thereby monitoring the whole production process, playing the role of omnibearing quality monitoring, realizing the automatic control of the butyric acid fermentation process, simultaneously reducing the labor cost, reducing the human error, ensuring the consistency of batch processes and ensuring the reliable product quality.
Description
Technical Field
The invention belongs to the technical field of near infrared spectroscopy, and particularly relates to a method for controlling butyric acid fermentation by using online near infrared.
Background
The biological fermentation industry is one of the support industries in China. However, the current industry development has many problems, such as low detection level of fermentation process parameters, unpopular automation degree and the like. Particularly, when various indexes are detected in the fermentation process, different instruments are required to be used for detection according to different indexes, for example, a pH measuring instrument is required for detecting the pH value, a spectrophotometer is required for detecting the bacterial concentration, a glucose measuring instrument is required for detecting the glucose content, corresponding chemical reactions are required for detecting the acid yield, and the like.
With the progress of science and technology and the development of detection equipment, the analysis of the fermentation process in the future emphasizes the real-time automatic monitoring of various key process parameters, the statistics of analysis data results and the feedback control of the process.
The Near Infrared (NIR) is an electromagnetic wave between the ultraviolet visible light and the mid-infrared light, and has a wavelength range of 780-2526 nm. The NIR region reflects mainly the double frequency and combined frequency absorption of H-containing groups in the mid-infrared region. Therefore, the band is very suitable for measuring the physical and chemical parameters of the organic compounds. In addition, NIR spectroscopy is rich in material structural and compositional information and can therefore be used for qualitative and quantitative analysis. At present, the near infrared technology is applied to the industries of traditional Chinese medicine, feed, grain and oil, food and the like in the reports of documents, but the near infrared technology is not applied to butyric acid fermentation.
Chinese patent CN102023140A discloses a method for measuring the content of 1-2 propylene glycol by NIR technology, which uses the near infrared spectrum technology to measure 1-2 propylene glycol, but the measured substance does not relate to the change of the biological fermentation process, and the measured components are less; in addition, Chinese patent CN 105548064A discloses a method for measuring various nutrient components and antibiotic titer in the antibiotic fermentation process by using an NIR technology, but the method disclosed in the patent is used for measuring related substance indexes off line, has certain hysteresis, still needs manual operation, and cannot realize real-time monitoring and automatic control of the fermentation process.
Due to the complexity, nonlinearity and time-varying nature of biological processes, process detection and control systems based on physical and chemical sensors have not been able to meet the needs of actual production, and many optimization models and automatic control systems have not been well implemented in actual production. One of the reasons is the lack of rapid detection or on-line detection technology for the most direct control parameters of the production process, namely biological (biochemical) parameters (including substrates, important intermediate metabolites, target products and the like), which is also a bottleneck technology faced by the whole industry.
With the rapid development of modern industry, the production equipment and scale required by the fermentation industry are continuously expanding. Intelligent and automatic manufacturers have more and more urgent requirements on automatic control technology, and the quality control of the fermentation industry emphasizes on online monitoring of quality-related factors in the production process, such as pH, bacterial liquid concentration, glucose content, product concentration and other monitoring parameters in the fermentation process. Therefore, there is a strong need for a more convenient and fast online measurement method to be applied to the fermentation industry.
Disclosure of Invention
In view of the above, the present invention provides a method for controlling butyric acid fermentation by using online near infrared, which uses an online near infrared spectroscopy technology to automatically monitor, count and feed back parameters of the butyric acid anaerobic fermentation process in real time, so as to realize automatic control of the butyric acid fermentation process, reduce labor cost, reduce human errors, ensure consistent batch-to-batch processes, and ensure reliable product quality.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for controlling butyric acid fermentation by using online near infrared, which comprises the following steps: (1) carrying out continuous online detection in the process of producing butyric acid by microbial fermentation, obtaining the near infrared spectrum of each sample by adopting an indium gallium arsenic diode array mode and continuous grating full-wavelength scanning during detection, simultaneously sampling every 4h and measuring the process parameter index of each sample by using a standard method to obtain detection data; the process parameter indexes comprise glucose content, biomass and butyric acid content;
(2) performing multivariate scattering correction pretreatment, first-order derivative pretreatment, smoothing pretreatment or vector normalization pretreatment on the collected near infrared spectrum data by using computer software and a chemometric method according to the collected spectral data of the fermentation liquor, and calculating by combining a principal component analysis method and a partial least square method to obtain a fitting equation of the near infrared spectrum data and physicochemical detection data, namely establishing a fermentation liquor model; and detecting the parameter content and the variation trend in the microbial fermentation process by using the model.
Preferably, the microorganism of step (1) is clostridium butyricum; the fermentation is anaerobic fermentation.
Preferably, the wavelength range of the near infrared spectrum in the step (1) is selected from 950-1650 nm; the spot diameter was 100 mm.
Preferably, after the fermentation broth model is established in the step (2), the method further comprises evaluating two indexes of a cross validation correlation coefficient and a cross validation standard deviation, and determining the model with the relatively largest cross validation correlation coefficient and the relatively smallest cross validation standard deviation as the final butyric acid fermentation broth model.
Preferably, the final butyric acid fermentation broth model is used for realizing the online measurement of metabolic parameters in the butyric acid production process of the anaerobic fermentation of clostridium butyricum, and the feeding rate is adjusted according to the parameter change to realize the online control.
The invention provides a method for controlling butyric acid fermentation by using online near infrared, which is characterized in that the production activity is subjected to feedback regulation by online detection of data and combination with actual production process conditions, so that the conversion rate of a substrate and the content of a target product are improved, the consumption of raw materials is reduced, and the economic benefit of an enterprise is increased.
Drawings
FIG. 1 is a graph showing data of a quality control manual detection method and an online near-infrared model prediction method for detecting glucose content in a butyric acid fermentation process;
FIG. 2 is detection data of butyric acid content in the butyric acid fermentation process by a quality control manual detection method and an online near-infrared model prediction method;
FIG. 3 shows the detection data of biomass in the butyric acid fermentation process by the quality control manual detection method and the online near-infrared model prediction method.
Detailed Description
The invention provides a method for controlling butyric acid fermentation by using online near infrared, which comprises the following steps: (1) carrying out continuous online detection in the process of producing butyric acid by microbial fermentation; in the detection process, an indium gallium arsenic diode array mode is adopted, the near infrared spectrum of each sample is obtained through continuous grating full-wavelength scanning, meanwhile, samples are taken every 4 hours, the process parameter index of each sample is measured by a standard method, and detection data are obtained; the process parameter indexes comprise glucose content, biomass and butyric acid content;
(2) performing multivariate scattering correction pretreatment, first-order derivative pretreatment, smoothing pretreatment or vector normalization pretreatment on the collected near infrared spectrum data by using computer software and a chemometric method according to the collected spectral data of the fermentation liquor, and calculating by combining a principal component analysis method and a partial least square method to obtain a fitting equation of the near infrared spectrum data and physicochemical detection data, namely establishing a fermentation liquor model; and detecting the parameter content and the variation trend in the microbial fermentation process by using the model.
The method comprises the steps of carrying out continuous online detection in the process of producing butyric acid by microbial fermentation, obtaining the near infrared spectrum of each sample by adopting an indium gallium arsenic diode array mode during detection and continuous grating full-wavelength scanning, sampling every 4 hours, and measuring the process parameter index of each sample by using a standard method to obtain detection data; the process parameter indexes comprise glucose content, biomass and butyric acid content. The microorganism of the invention is preferably clostridium butyricum; the fermentation is preferably anaerobic fermentation. The invention preferably carries out continuous on-line detection on the process of producing the butyric acid by the anaerobic fermentation of the clostridium butyricum, and the detected indexes comprise biomass, glucose content and butyric acid content.
The present invention preferably employs continuous grating full wavelength scanning using a model DA7440 on-line near infrared analyzer from waotong corporation. The wavelength range of the near infrared spectrum is preferably selected from 950-1650 nm; the spot diameter is preferably 100 mm.
The method comprises the steps of collecting spectral data of fermentation liquor, utilizing computer software and a chemometrics method to carry out multivariate scattering correction pretreatment, first-order derivative pretreatment, smoothing pretreatment or vector normalization pretreatment on the collected near infrared spectral data, combining a principal component analysis method and a partial least square method to calculate a fitting equation of the near infrared spectral data and physicochemical detection data, namely establishing a fermentation liquor model, utilizing the model to detect parameter content and variation trend in the microbial fermentation process, collecting spectral data of the fermentation liquor, preferably carrying out regression calculation on the spectral data and the chemical analysis detection data through chemometrics software, determining a functional relation, then establishing a near infrared model, carrying out continuous online scanning on the fermentation liquor when parameters of the fermentation process of the clostridium butyricum are determined, utilizing the established near infrared model according to the spectral data of the fermentation liquor at different time points, calculating the parameter content and the variation trend in the whole fermentation process of the clostridium butyricum, preferably further comprising the steps of carrying out the continuous online scanning on the fermentation liquor after the fermentation process parameters of the clostridium butyricum fermentation are determined, selecting the near infrared model to carry out the cross validation of two indexes of the related coefficients, the cross validation of the maximum and the cross validation of the variance of the fermentation of the clostridium butyricum in the whole fermentation process, selecting the anaerobic fermentation of the clostridium butyricum, adjusting the anaerobic fermentation of the clostridium butyricum, and the anaerobic fermentation of the butyric acid, and the anaerobic fermentation of.
The method for controlling butyric acid fermentation by on-line near infrared according to the present invention will be described in detail with reference to the following examples, which should not be construed as limiting the scope of the present invention.
Example 1
1) Detecting metabolic parameters in the process of producing butyric acid by the anaerobic fermentation of clostridium butyricum;
2) the method comprises the steps of utilizing a continuous grating full-wavelength scanning of a Doutong company DA7440 type on-line near-infrared analyzer to obtain the near-infrared spectrum of each sample on line, processing all spectrum information, and manually measuring the process parameter index of each sample through a quality control unit (QC) of the company every 4 hours to obtain detection data. The process parameter indexes comprise biomass, glucose content and butyric acid content. The wavelength range of the near infrared spectrum is selected from 950-1650 nm.
3) The parameter method for manually detecting the fermentation process of clostridium butyricum by the tasting tube part comprises the following steps:
biomass determination: determination of OD Using Spectrophotometer600。
And (3) measuring the content of glucose: the measurement was carried out using a glucose meter.
And (3) determination of butyric acid content: the determination is carried out by high performance liquid chromatography.
4) Establishing a near-infrared fermentation liquor detection model: the method specifically comprises the following steps: performing multivariate scattering correction pretreatment, first derivative pretreatment, smoothing pretreatment or vector normalization pretreatment on the collected near infrared spectrum data by using computer software and a chemometric method, and calculating by combining a principal component analysis method and a partial least square method to obtain a fitting equation of the near infrared spectrum data and physicochemical detection data, namely establishing a fermentation liquid model; and then, through evaluation of two indexes of the cross validation correlation coefficient and the cross validation standard deviation, selecting a model with a relatively large cross validation correlation coefficient and a relatively small cross validation standard deviation to determine the model as the final butyric acid fermentation liquor model. The final butyric acid fermentation broth model is utilized to realize the online measurement of metabolic parameters in the process of producing butyric acid by the anaerobic fermentation of clostridium butyricum, and the feed supplement rate is adjusted according to the parameter change to realize the online control. The modeling data is shown in table 1:
TABLE 1 Clostridium butyricum fermentation broth sample modeling data case
(5) the final butyric acid fermentation liquor model is adopted to realize the on-line measurement of metabolic parameters in the butyric acid production process of the anaerobic fermentation of clostridium butyricum, and the comparison and verification of physicochemical detection data of a quality control department are carried out, the results are shown in figures 1-3, the data obtained by the quality control manual detection method and the on-line near infrared model prediction method are consistent no matter the content of butyric acid or the content of glucose and biomass, the artificial detection can be replaced, and the yield of butyric acid reaches more than 50 g/L.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (5)
1. A method for controlling butyric acid fermentation by using online near infrared is characterized by comprising the following steps: (1) carrying out continuous online detection in the process of producing butyric acid by microbial fermentation; during the online detection, a near infrared spectrum of each sample is obtained by adopting an indium gallium arsenide diode array mode and continuous grating full-wavelength scanning, and meanwhile, samples are taken every 4 hours, and process parameter indexes of each sample are measured by a standard method to obtain detection data; the process parameter indexes comprise glucose content, biomass and butyric acid content;
(2) performing multivariate scattering correction pretreatment, first-order derivative pretreatment, smoothing pretreatment or vector normalization pretreatment on the collected near infrared spectrum data by using computer software and a chemometric method according to the collected spectral data of the fermentation liquor, and calculating by combining a principal component analysis method and a partial least square method to obtain a fitting equation of the near infrared spectrum data and physicochemical detection data, namely establishing a fermentation liquor model; and detecting the parameter content and the variation trend in the microbial fermentation process by using the model.
2. The method according to claim 1, wherein the microorganism of step (1) is Clostridium butyricum; the fermentation is anaerobic fermentation.
3. The method according to claim 1, wherein the wavelength range of the near infrared spectrum in the step (1) is selected from 950 to 1650 nm; the spot diameter was 100 mm.
4. The method according to claim 1, wherein after the fermentation broth model is established in step (2), the method further comprises the step of selecting the model with the relatively largest cross validation correlation coefficient and the relatively smallest cross validation standard deviation as the final butyric acid fermentation broth model through evaluation of two indexes of the cross validation correlation coefficient and the cross validation standard deviation.
5. The method according to claim 4, wherein the final butyric acid fermentation broth model is used for realizing online measurement of metabolic parameters in the butyric acid production process of the anaerobic fermentation of clostridium butyricum, and the feeding rate is adjusted according to parameter changes to realize online control.
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| CN114486799A (en) * | 2022-04-15 | 2022-05-13 | 广东省农业科学院动物科学研究所 | Detection method and system for clostridium butyricum in poultry intestinal tract |
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