WO2022127203A1 - Appareil et procédé de mesure de polysaccharides dans un bouillon de fermentation en fonction des spectres raman, et application de l'appareil et du procédé - Google Patents

Appareil et procédé de mesure de polysaccharides dans un bouillon de fermentation en fonction des spectres raman, et application de l'appareil et du procédé Download PDF

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Publication number
WO2022127203A1
WO2022127203A1 PCT/CN2021/116536 CN2021116536W WO2022127203A1 WO 2022127203 A1 WO2022127203 A1 WO 2022127203A1 CN 2021116536 W CN2021116536 W CN 2021116536W WO 2022127203 A1 WO2022127203 A1 WO 2022127203A1
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WIPO (PCT)
Prior art keywords
fermentation broth
optical fiber
polysaccharides
chip microcomputer
detector
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PCT/CN2021/116536
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English (en)
Chinese (zh)
Inventor
史建国
李秋顺
杨俊慧
马耀宏
蔡雷
杨艳
刘庆艾
孟庆军
王丙莲
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山东省科学院生物研究所
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Publication of WO2022127203A1 publication Critical patent/WO2022127203A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/65—Raman scattering
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • 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
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • 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
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N2021/0106—General arrangement of respective parts
    • G01N2021/0112—Apparatus in one mechanical, optical or electronic block
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/65—Raman scattering
    • G01N2021/653—Coherent methods [CARS]
    • G01N2021/655—Stimulated Raman

Definitions

  • the invention belongs to the technical field of fermentation broth detection, and in particular relates to a device for detecting polysaccharides in fermentation broth based on Raman spectroscopy, a method and application thereof.
  • polysaccharides have unique physical and chemical properties and biological activities, and are a kind of very promising active substances.
  • polysaccharides have been used in many fields such as food, medicine, light industrial production, beauty, petroleum, medical treatment, etc., and more and more people's attention has been paid, and it has a wider application prospect.
  • the content of polysaccharide in the fermentation broth is a marker for judging the success of fermentation and an important indicator for judging strains and optimizing fermentation conditions. Therefore, the accurate determination of polysaccharide content is particularly important for the process control of fermentation.
  • most of the devices for detecting polysaccharides in fermentation broth are to take out the fermentation broth from the fermentation tank, and then process it through biological or chemical reactions to obtain the content of polysaccharides, mainly including HPLC method, ELISA method, and dinitrosalicylic acid method. , phenol-sulfuric acid method, etc.
  • the HPLC detection device has high precision, but this method takes too long and the pretreatment is troublesome; the ELISA detection method has a low detection limit and high sensitivity, but the detection steps of this method are cumbersome and require multiple washings; The determination steps of the dinitrosalicylic acid method and the phenol-sulfuric acid method are very complicated. These traditional assay methods cannot reflect the components in the fermentation broth in real time, and the detection results have serious lag. Simple, fast and suitable for the requirements of on-site rapid detection, it is an inevitable trend of the development of detection methods in the future. Therefore, it is necessary to find a method for real-time online detection of polysaccharides in fermentation broth.
  • the present invention provides a device for detecting polysaccharides in fermentation broth based on Raman spectroscopy, which can realize on-line detection of polysaccharides in fermentation broth and has good practical application value.
  • a first aspect of the present invention provides a device for detecting polysaccharides in fermentation broth based on Raman spectroscopy, the device comprising a fermenter, a reflector, an optical fiber probe, an optical fiber, an excitation light source, a detector, a signal acquisition module, a D conversion module, microcontroller and display module.
  • the reflector is located in the fermenter, the excitation light source is connected to the fiber probe through an optical fiber, and the fiber probe is connected to the detector through an optical fiber.
  • the detector is further connected to the signal acquisition module, and then the signal acquisition module is connected to the A/D conversion module.
  • the A/D conversion module is connected with the single-chip microcomputer, and the single-chip microcomputer is connected with the display module.
  • a second aspect of the present invention provides a method for detecting polysaccharides in fermentation broth based on Raman spectroscopy, the method comprising detecting polysaccharides in fermentation broth based on the above device.
  • the detection method includes:
  • the detector transmits the detected Raman spectrum signal to the signal acquisition module
  • the single-chip microcomputer performs algorithm processing on the digital signal, and separates and calculates the spectrum of pure polysaccharide and the content of polysaccharide;
  • the display module can display the Raman spectrum of pure polysaccharide and/or the content value of polysaccharide.
  • the third aspect of the present invention provides the application of the above-mentioned device and/or method for detecting polysaccharides in fermentation broth based on Raman spectroscopy in detecting the content or purity of polysaccharides in fermentation broth.
  • the device for detecting polysaccharides in fermentation broth based on Raman spectroscopy provided by the above technical solution does not require purification and separation, simplifies detection steps, and saves detection time.
  • the device for detecting polysaccharides in fermentation broth based on Raman spectroscopy provided by the above technical solution does not need to sample and then detect, and can detect polysaccharides online in real time, effectively solving the shortcoming of serious lag in detection results.
  • the device for detecting polysaccharides in fermentation broth based on Raman spectroscopy provided by the above technical solution does not need to undergo complex biological or chemical reaction treatment, thereby reducing the detection cost of polysaccharides.
  • the device for detecting polysaccharides in fermentation broth based on Raman spectroscopy provided by the above technical solution has broad commercial application prospects in the field of fermentation industry, and is expected to be popularized and applied on a large scale.
  • FIG. 1 is a schematic diagram of the process of detecting polysaccharides in fermentation broth based on Raman spectroscopy in an embodiment of the present invention.
  • Fermentation tank 2. Reflector, 3. Optical fiber probe, 4. Optical fiber, 5. Optical fiber, 6. Excitation light source, 7. Detector, 8. Signal acquisition module, 9. A/D conversion module, 10 , SCM, 11, LCD screen, 12, mobile phone, 13, computer.
  • orientation or positional relationship is based on the orientation or positional relationship shown in the accompanying drawings, and is only a relational word determined for the convenience of describing the structural relationship of each component or element of the present invention, and does not specifically refer to any component or element in the present invention, and should not be construed as a reference to the present invention. Invention limitations.
  • a device for detecting polysaccharides in fermentation broth based on Raman spectroscopy includes a fermenter, a reflector, an optical fiber probe, an optical fiber, an excitation light source, a detector, a signal Acquisition module, A/D conversion module, microcontroller and display module.
  • the reflector is located in the fermenter, the excitation light source is connected to the fiber probe through an optical fiber, and the fiber probe is connected to the detector through an optical fiber.
  • the detector is further connected to the signal acquisition module, and then the signal acquisition module is connected to the A/D conversion module.
  • the A/D conversion module is connected with the single-chip microcomputer, and the single-chip microcomputer is connected with the display module.
  • the reflector is fixed on the bottom side of the fermentation tank
  • the mirror surface of the reflector is perpendicular to the incident light of the fiber probe
  • the mirror surface of the reflector is a smooth plane or a mirror surface composed of micro-nano structures such as triangular, cylindrical, square, pentagonal, hexagonal, etc.;
  • the optical fiber probe is located inside the fermentation liquid, and monitors the components in the fermentation liquid in situ in real time;
  • the detector is a Raman spectroscopic detector, not other types of spectroscopic detectors; therefore, the collected signals of the components in the fermentation broth are Raman spectroscopic signals.
  • the display module includes but is not limited to any one or more of a liquid crystal display screen, a mobile phone and a computer;
  • the single-chip microcomputer and the display module can be connected by any one or more of data lines, bluetooth, WIFI or USB.
  • a method for detecting polysaccharides in fermentation broth based on Raman spectroscopy comprising detecting polysaccharides in fermentation broth based on the above device.
  • the detection method includes:
  • the detector transmits the detected Raman spectrum signal to the signal acquisition module
  • the single-chip microcomputer performs algorithm processing on the digital signal, and separates and calculates the spectrum of pure polysaccharide and the content of polysaccharide;
  • the display module includes but is not limited to any one or more of a liquid crystal display screen, a mobile phone and a computer;
  • the single-chip microcomputer and the display module can be connected by any one or more of a data cable, Bluetooth, WIFI or USB.
  • the display module can display the Raman spectrum of pure polysaccharide and/or the content value of polysaccharide.
  • the application of the above-mentioned device and/or method for detecting polysaccharides in fermentation broth based on Raman spectroscopy is provided in detecting the content or purity of polysaccharides in fermentation broth.
  • a device for detecting polysaccharides in fermentation broth based on Raman spectroscopy comprising a fermenter (1), a mirror (2), an optical fiber probe (3), optical fibers (4, 5), and an excitation light source (6) ), a detector (7), a signal acquisition module (8), an A/D conversion module (9), a single-chip microcomputer (10), a liquid crystal display (11), a mobile phone (12) or a computer (13); (2) is located in the fermenter (1), the excitation light source (6) is connected to the optical fiber probe (3) through the optical fiber (4), and the optical fiber probe (3) is further connected to the detector (7) through the optical fiber (5), and the detector (7) is further connected with the signal acquisition module (8), then the signal acquisition module (8) is connected with the A/D conversion module (9), and the A/D conversion module (9) is connected with the further single chip (10), The single-chip microcomputer (10) is connected with the liquid crystal display screen (11), the mobile phone (12) or the computer (13) through a data cable
  • the reflecting mirror (2) is fixed on the bottom part of the fermentation tank (1).
  • the mirror surface of the reflecting mirror (2) is perpendicular to the incident light of the optical fiber probe (3).
  • the mirror surface of the reflecting mirror (2) is a smooth plane or a mirror surface composed of micro-nano structures such as triangular, cylindrical, square, pentagonal, and hexagonal.
  • the optical fiber probe (3) is located inside the fermentation liquid, and monitors the components inside the fermentation liquid in real time and in situ.
  • the detector (7) is a Raman spectral detector, not other types of spectral detectors.
  • the collected signals of components in the fermentation broth are Raman spectrum signals.
  • the Raman spectral data obtained by the detector (7) is further processed by an algorithm to separate the Raman spectral signal of the polysaccharide component.
  • a device for detecting polysaccharides in fermentation broth based on Raman spectroscopy the detection process is as follows:
  • the detector (7) transmits the detected Raman spectrum signal to the signal acquisition module (8);
  • the single chip microcomputer (10) performs algorithm processing on the digital signal, separates and calculates the spectrum of pure polysaccharide and the content of polysaccharide;
  • the signals and data processed by the single chip microcomputer (10) are transmitted through a data cable or bluetooth or WIFI or USB, and the Raman of pure polysaccharide is stored and displayed on the liquid crystal display screen (11) or mobile phone (12) or computer (13). Spectral and polysaccharide content values.

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  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

La présente invention se rapporte au domaine technique de l'analyse des bouillons de fermentation. L'invention concerne un appareil et un procédé pour mesurer les polysaccharides dans un bouillon de fermentation en fonction des spectres Raman, et l'application de l'appareil et du procédé. L'appareil de mesure de polysaccharides dans un bouillon de fermentation en fonction de spectres Raman comprend une cuve de fermentation, un miroir réfléchissant, une sonde à fibre optique, une fibre optique, une source de lumière d'excitation, un détecteur, un module d'acquisition de signaux, un module de conversion analogique-numérique, un micro-ordinateur monopuce et un module d'affichage, le miroir réfléchissant se trouvant à l'intérieur de la cuve de fermentation ; la source de lumière d'excitation est reliée à la sonde à fibre optique au moyen de la fibre optique, et la sonde à fibre optique est reliée au détecteur au moyen de la fibre optique ; le détecteur est en outre relié au module d'acquisition de signaux ; le module d'acquisition de signaux est relié au module de conversion analogique-numérique ; le module de conversion analogique-numérique est relié au micro-ordinateur monopuce ; et le micro-ordinateur monopuce est relié au module d'affichage. Par rapport à un procédé traditionnel de mesure de polysaccharides, l'appareil de la présente invention ne nécessite pas une purification et une séparation, ce qui simplifie les étapes de mesure, permet de gagner du temps de mesure et de réduire les coûts de mesure des polysaccharides. La présente invention présente une grande valeur d'application pratique.
PCT/CN2021/116536 2020-12-17 2021-09-03 Appareil et procédé de mesure de polysaccharides dans un bouillon de fermentation en fonction des spectres raman, et application de l'appareil et du procédé WO2022127203A1 (fr)

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CN202011495734.3A CN112730376A (zh) 2020-12-17 2020-12-17 一种基于拉曼光谱检测发酵液中多糖的装置及其方法和应用
CN202011495734.3 2020-12-17

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Cited By (1)

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WO2024061126A1 (fr) * 2022-09-23 2024-03-28 北京蓝晶微生物科技有限公司 Procédé, appareil et système d'évaluation de teneur en polyhydroxyalcanoate, et dispositif

Families Citing this family (1)

* Cited by examiner, † Cited by third party
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CN112730376A (zh) * 2020-12-17 2021-04-30 山东省科学院生物研究所 一种基于拉曼光谱检测发酵液中多糖的装置及其方法和应用

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CN111344558A (zh) * 2017-10-06 2020-06-26 龙沙有限公司 使用拉曼光谱法自动控制细胞培养
CN112730376A (zh) * 2020-12-17 2021-04-30 山东省科学院生物研究所 一种基于拉曼光谱检测发酵液中多糖的装置及其方法和应用

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CN104483287A (zh) * 2014-12-05 2015-04-01 南京工业大学 基于近红外光谱的在线发酵过程生物学参数的检测装置及方法
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CN111323407A (zh) * 2020-02-20 2020-06-23 浙江大学 一种快速测定中药多糖含量的拉曼光谱检测方法
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024061126A1 (fr) * 2022-09-23 2024-03-28 北京蓝晶微生物科技有限公司 Procédé, appareil et système d'évaluation de teneur en polyhydroxyalcanoate, et dispositif

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