CN115246880A - Novel method for greatly improving phycocyanin content - Google Patents
Novel method for greatly improving phycocyanin content Download PDFInfo
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- CN115246880A CN115246880A CN202110465277.1A CN202110465277A CN115246880A CN 115246880 A CN115246880 A CN 115246880A CN 202110465277 A CN202110465277 A CN 202110465277A CN 115246880 A CN115246880 A CN 115246880A
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- 108010053210 Phycocyanin Proteins 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 21
- 240000002900 Arthrospira platensis Species 0.000 claims abstract description 44
- 235000016425 Arthrospira platensis Nutrition 0.000 claims abstract description 44
- 229940082787 spirulina Drugs 0.000 claims abstract description 44
- 239000012528 membrane Substances 0.000 claims abstract description 36
- 239000011344 liquid material Substances 0.000 claims abstract description 17
- 238000001514 detection method Methods 0.000 claims abstract description 10
- 241001474374 Blennius Species 0.000 claims abstract description 6
- 230000001954 sterilising effect Effects 0.000 claims abstract description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 3
- 239000011550 stock solution Substances 0.000 claims abstract description 3
- 210000004027 cell Anatomy 0.000 claims description 19
- 210000000170 cell membrane Anatomy 0.000 claims description 12
- 239000004065 semiconductor Substances 0.000 claims description 10
- 238000004891 communication Methods 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 3
- 238000012544 monitoring process Methods 0.000 claims description 3
- 230000002572 peristaltic effect Effects 0.000 claims description 3
- 238000000605 extraction Methods 0.000 abstract description 3
- 230000006870 function Effects 0.000 description 6
- 238000010586 diagram Methods 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- 239000013307 optical fiber Substances 0.000 description 4
- 239000003990 capacitor Substances 0.000 description 3
- 238000004520 electroporation Methods 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 230000002093 peripheral effect Effects 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000005684 electric field Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000206572 Rhodophyta Species 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000005907 cancer growth Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 238000003487 electrochemical reaction Methods 0.000 description 1
- 238000004146 energy storage Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000004952 protein activity Effects 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/795—Porphyrin- or corrin-ring-containing peptides
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/06—Lysis of microorganisms
- C12N1/066—Lysis of microorganisms by physical methods
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a novel method for greatly improving the content of phycocyanin.A device comprises a high-voltage pulse generator, a spirulina membrane breaking treatment chamber, a liquid material conveying system and a control and detection system; the high-voltage pulse generator has the function of generating high-voltage pulses, and the generated high-voltage pulses act on the electrode of the spiral seaweed membrane breaking treatment chamber; the spirulina membrane breaking treatment chamber is a place for releasing phycocyanin after the spirulina membrane is broken; the liquid material conveying system is connected with one side of the spirulina membrane breaking treatment chamber and is used for conveying spirulina stock solution to the treatment chamber; the control and detection system is mainly responsible for maintaining the normal operation of the high-voltage pulse sterilization equipment. The device disclosed by the invention is low in complexity and small in size, and the implementation method can reduce the loss of phycocyanin in the extraction process and greatly improve the content of the extracted phycocyanin.
Description
Technical Field
The invention relates to the field of food processing, in particular to a novel method for greatly improving the content of phycocyanin.
Background
Phycocyanin is a dark blue powder isolated from spirulina. Mainly present in spirulina, red algae and cryptophyceae. Phycocyanin can help regulate and synthesize various important enzymes required by human metabolism, has important functions of inhibiting the growth of cancer cells, promoting the regeneration of human cells, maintaining ovaries and promoting the synthesis of elastin in human bodies, and simultaneously regulates the immune system of human bodies, enhances the functions of the immune system and improves the resistance of the human bodies to diseases. Currently, a freeze-thawing method is generally adopted for extracting spirulina cell sap: freezing the cells at low temperature for a certain time, taking out the cells, thawing the cells at room temperature, repeating the freezing and thawing for multiple times, and swelling and breaking the cells while forming ice particles and increasing the concentration of the residual cytosol salt. Although simple and convenient, the freeze-thaw method is time-consuming and labor-consuming and is prone to damage to protein activity. Therefore, a new method for extracting spirulina cell sap needs to be searched.
Disclosure of Invention
The invention aims to provide a novel method for greatly improving the content of phycocyanin, and provides a device for electroporation of cells based on high-energy pulses, which can quickly extract cell sap in spirulina and reduce the damage to phycocyanin in the cell sap as much as possible.
In order to achieve the purpose, the invention adopts the technical scheme that:
a novel method for greatly improving the content of phycocyanin is characterized in that: comprises a high-voltage pulse generator, a spirulina membrane-breaking treatment chamber, a control and detection system and a liquid material conveying system; the high-voltage pulse generator has the function of generating high-voltage pulses, and the generated high-voltage pulses act on the electrode of the spirulina membrane breaking treatment chamber; the spirulina membrane-breaking treatment chamber is a place for releasing phycocyanin after the spirulina membrane is broken; the liquid material conveying system is connected with one side of the spirulina membrane breaking treatment chamber and is used for conveying spirulina stock solution to the treatment chamber; the control and detection system is mainly responsible for maintaining the normal operation of the high-voltage pulse sterilization equipment.
As a further improvement of the invention, the high-voltage pulse generator is provided with a direct-current high-voltage source control platform and a step-up transformer, and converts the input 220V alternating current into high-voltage direct current.
As a further improvement of the invention, the spirulina membrane-breaking treatment chamber utilizes the generated high-voltage pulse to break down the cell membrane of the spirulina conveyed by the liquid material conveying system, so that the phycocyanin in the spirulina is released out of the cells.
As a further improvement of the invention, the control and detection system comprises a power supply circuit, an FPGA + STM32 interface circuit, an RS232 and RS485 communication circuit, a pulse output circuit and an AD acquisition circuit, and the control of the output high-voltage pulse is realized through AD acquisition data and set original parameters.
As a further improvement of the invention, the liquid material conveying system is composed of a peristaltic pump, and the spirulina material to be treated is conveyed into the spirulina membrane breaking treatment chamber and is cooled and collected after membrane breaking is finished.
The invention relates to a novel method for greatly improving the content of phycocyanin, which is characterized by comprising the following specific steps:
a) The high-voltage pulse generator is characterized in that a standard alternating voltage of 220V is converted into an alternating current of dozens of kV through an inverter transformer, then the alternating current is converted into a high-voltage direct voltage through rectification, and an exponential decay wave is generated through a high-voltage semiconductor switch. (ii) a
b) A spirulina membrane breaking treatment chamber for breaking the membrane of the spirulina by generating high-voltage pulse;
c) Sending a pulse trigger signal to control a high-voltage semiconductor switch driver and a liquid material conveying system to convey materials to the spiral seaweed membrane breaking treatment chamber, and completing acquisition control and detection systems for various monitoring parameters of the spiral seaweed membrane breaking treatment chamber;
d) And the liquid material conveying system is used for conveying the spirulina material to be subjected to membrane rupture into the spirulina membrane rupture treatment chamber and cooling and collecting the spirulina material after membrane rupture.
Compared with the prior art, the invention has the beneficial effects that:
1) Aiming at the condition that the traditional phycocyanin extraction method is time-consuming and labor-consuming, the invention utilizes the phenomenon and mechanism of electroporation of high-energy Pulse (PEF) to cells and adopts the high-energy pulse to break the cell membrane of spirulina, thereby not only reducing the cost and greatly reducing the equipment volume, but also reducing the phycocyanin loss in the extraction process;
2) The invention designs a main control board taking STM32F103ZET6 as a core, and the main control board finishes the following functions: the FPGA is controlled to output pulse control signals to the optical fiber transmitter, and the pulse control signals are transmitted to the high-voltage semiconductor switch driver through the optical fiber, so that the stability and the safety of the system are greatly improved;
3) The invention designs a high-voltage semiconductor switch driver taking a CPLD as a core to drive the on-off of a high-voltage semiconductor switch, and different high-isolation DC-DC voltage conversions can be carried out by modifying the circuit structure of two circuits.
Drawings
FIG. 1 is a diagram showing the electroporation process of cell membranes in the present invention.
Fig. 2 is a circuit diagram of the main control board hardware according to the present invention.
Fig. 3 is a hardware circuit structure diagram of the high voltage semiconductor switch driver according to the present invention.
Fig. 4 is a schematic circuit diagram of a bipolar high voltage pulse generator according to the present invention.
FIG. 5 is a block diagram of a control structure of the liquid conveying system of the present invention.
Detailed Description
The present invention will be further illustrated with reference to the accompanying drawings and specific embodiments, which are to be understood as merely illustrative of the invention and not as limiting the scope of the invention.
As shown in FIG. 1, since the cell membrane of the microorganism is sensitive to the electric field, when a pulsed electric field of a certain intensity is applied to the cell membrane, a potential difference is formed between the inside and the outside of the cell membrane, and a series of electrochemical reactions are induced. These reactions change the properties and structure of the cell membrane phospholipid bilayer, resulting in several-fold enlargement of the original membrane pores of the cell membrane and formation of new membrane pores with better hydrophobicity. Along with the increase of large membrane pores, the permeability of the cell membrane is gradually improved, a plurality of small molecular substances (mainly water molecules) outside the cell penetrate through the cell membrane to enter the interior of the cell, the cell volume is gradually increased, and finally the cell membrane is expanded and broken, and substances in the cell are leaked out, so that the aim of obtaining the cell sap of the spirulina cell is achieved.
As shown in fig. 2, the main control board hardware circuit mainly includes a power supply circuit, an FPAG + STM32 interface circuit, an RS232 and RS485 communication circuit, an AD acquisition circuit, a pulse output circuit, and a relay output circuit. The power supply circuit is used for supplying power to each module circuit in the main control board. The FPAG + STM32 interface circuit realizes communication with the FPGA by adopting an STM32F103ZET6 variable static memory controller (FSMC) parallel bus interface. The RS232 and RS485 communication circuits are connected with the STM32, and the RS232 serial port is used for communicating with the embedded monitoring platform of the human-computer interaction equipment; and the RS485 serial port is used for communicating with the liquid material conveying system. And the AD acquisition circuit is connected to the STM32 and is used for acquiring the temperature, the pressure and the flow in the spirulina membrane breaking treatment chamber when the system runs. The pulse output circuit is controlled by the FPGA and converts the pulse control electric signal into an optical signal in an optical fiber transmission mode and sends the optical signal to the switch driver. And the relay output circuit is controlled by the STM32 to control the power supply of the whole system, the start and stop of the refrigeration compressor, and the switch and alarm lamp output of the equipment cabinet door.
As shown in fig. 3, the high-voltage semiconductor switch driver hardware circuit includes a CPLD and its peripheral circuit, a power supply circuit, a pulse input circuit, and an IGBT driving circuit. The CPLD and the peripheral circuit thereof are used for processing the pulse control signal. The power supply circuit adopts a chip AMS1117CD-3.3 to step down to obtain +3.3V, and the +3.3V is subjected to capacitance filtering and then supplies power to the pulse input circuit and the CPLD basic peripheral circuit. The pulse input circuit has the main functions of receiving a pulse control signal sent by the FPGA through the optical fiber transmitter, converting a transmitted optical signal into an electric signal, realizing the conversion of signal electricity-light-electricity, and receiving and processing the signal by the CPLD. The IGBT driving circuit carries out level conversion on the pulse control signal, converts 3.3V in the pulse into +15V and converts 0V into-10V, and improves the driving capability of the IGBT driving circuit so as to smoothly control the on-off of the IGBT.
As shown in fig. 4, a bipolar high voltage pulse generator circuit is shown. The DC high-voltage source control platform outputs adjustable AC voltage of 0-220V, the DC high-voltage source is a step-up transformer, and the maximum step-up multiple can reach 300 times. For the positive polarity pulse power supply of the upper half part, the output voltage of the control platform is boosted through the boosting transformer to charge the energy storage capacitor C1, and R1 is a charging buffer resistor. The high-voltage semiconductor switch is the most critical device of a pulse power supply, the rising edge time and the falling edge time of output pulses are influenced by the on-off time of the switch to a great extent, the reliable switch also often determines the service life of the high-voltage pulse generator, the actual circuit of the high-voltage pulse generator is formed by connecting a plurality of IGBTs in series, and the on-off of the high-voltage pulse generator is controlled by a pulse signal provided by a switch driver. R4 and L1 are resistance and inductance of the circuit, the spiral seaweed membrane breaking treatment chamber has impedance characteristic and capacitive reactance characteristic, so the equivalent is that a capacitor C3 is connected with a resistor R3 in parallel, in order to prevent LC resonance generated by the series connection of the inductor L1 and the capacitor C3 from influencing pulse waveform, a freewheeling diode D1 is connected in parallel at two ends of the inductor L1, and the negative pulse power supply of the lower half part is in the same way.
As shown in fig. 5, the liquid material conveying system is controlled by the STM32, the main controller STM32 is used as a host to communicate with the peristaltic pumps of the slave machines through the RS485 communication interface according to the MODBUS communication protocol, and the slave machines realize corresponding functions in the commands after receiving the commands.
Claims (6)
1. A novel method for greatly improving the content of phycocyanin is characterized in that: comprises a high-voltage pulse generator, a spirulina membrane-breaking treatment chamber, a control and detection system and a liquid material conveying system; the high-voltage pulse generator has the function of generating high-voltage pulses, and the generated high-voltage pulses act on the electrode of the spirulina membrane breaking treatment chamber; the spirulina membrane breaking treatment chamber is a place for releasing phycocyanin after the membrane of the spirulina is broken; the liquid material conveying system is connected with one side of the spirulina membrane breaking treatment chamber and is used for conveying spirulina stock solution to the treatment chamber; the control and detection system is mainly responsible for maintaining the normal operation of the high-voltage pulse sterilization equipment.
2. The method of claim 1, wherein the method comprises the following steps: the high-voltage pulse generator is provided with a direct-current high-voltage source control platform and a step-up transformer and converts the input 220V alternating current into high-voltage direct current.
3. The method of claim 2, wherein the method comprises the following steps: the spirulina membrane breaking treatment chamber breaks down the spirulina cell membrane conveyed by the liquid material conveying system by utilizing the generated high-voltage pulse, so that the phycocyanin in the spirulina is released to the outside of the cell.
4. The method of claim 3, wherein the ratio of phycocyanin to phycocyanin is selected from the group consisting of: the control and detection system comprises a power supply circuit, an FPGA + STM32 interface circuit, RS232 and RS485 communication circuits, a pulse output circuit and an AD acquisition circuit, the control of output high-voltage pulses is realized through AD acquisition data and set original parameters, data display is realized through the RS232 communication circuit, and the control of the liquid material conveying system is realized through the RS485 communication circuit.
5. The method of claim 4, wherein the method comprises the following steps: the liquid material conveying system is composed of a peristaltic pump, the spirulina material to be treated is conveyed into the spirulina membrane breaking treatment chamber, and cooling and collection are carried out after membrane breaking is finished.
6. The novel method for greatly increasing the content of phycocyanin according to claim 4, comprising the following specific steps:
a) The high-voltage pulse generator is characterized in that a standard alternating voltage of 220V is converted into an alternating current of dozens of kV through an inverter transformer, then the alternating current is converted into a high-voltage direct voltage through rectification, and an exponential decay wave is generated through a high-voltage semiconductor switch;
b) A spirulina membrane breaking treatment chamber for breaking the membrane of the spirulina by generating high-voltage pulse;
c) Sending a pulse trigger signal to control a high-voltage semiconductor switch driver and a liquid material conveying system to convey materials to the spiral seaweed membrane breaking treatment chamber, and completing acquisition control and detection systems for various monitoring parameters of the spiral seaweed membrane breaking treatment chamber;
d) And the liquid material conveying system is used for conveying the spirulina materials to be subjected to membrane rupture into the spirulina membrane rupture treatment chamber and cooling and collecting the materials after membrane rupture.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110465277.1A CN115246880A (en) | 2021-04-28 | 2021-04-28 | Novel method for greatly improving phycocyanin content |
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| CN202110465277.1A CN115246880A (en) | 2021-04-28 | 2021-04-28 | Novel method for greatly improving phycocyanin content |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999049561A1 (en) * | 1998-03-24 | 1999-09-30 | The Ohio State Research Foundation | A high-voltage pulse generator |
| KR20000013474A (en) * | 1998-08-05 | 2000-03-06 | 박선순 | High electric fields sterilizing device and chamber for sterilization |
| CN211297060U (en) * | 2020-02-22 | 2020-08-18 | 南京林业大学 | Circuit for driving high-voltage pulse switch by FPGA |
| US20230174921A1 (en) * | 2020-04-29 | 2023-06-08 | Nofima As | Method And System Of Pre-Treating Biomass, In Particular Biomass That Is Resistant To Cell Disruption, For Improving The Accessibility Of Cellular Compounds Therefrom |
-
2021
- 2021-04-28 CN CN202110465277.1A patent/CN115246880A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999049561A1 (en) * | 1998-03-24 | 1999-09-30 | The Ohio State Research Foundation | A high-voltage pulse generator |
| KR20000013474A (en) * | 1998-08-05 | 2000-03-06 | 박선순 | High electric fields sterilizing device and chamber for sterilization |
| CN211297060U (en) * | 2020-02-22 | 2020-08-18 | 南京林业大学 | Circuit for driving high-voltage pulse switch by FPGA |
| US20230174921A1 (en) * | 2020-04-29 | 2023-06-08 | Nofima As | Method And System Of Pre-Treating Biomass, In Particular Biomass That Is Resistant To Cell Disruption, For Improving The Accessibility Of Cellular Compounds Therefrom |
Non-Patent Citations (6)
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| TOEPFL S, MATHYS A, HEINZ V, ET AL.: "Potential of high hydrostatic pressure and pulsed electric fields for energy efficient and environmentally friendly food processing", 《FOOD REVIEWS INTERNATIONAL》, vol. 22, no. 4, 6 February 2007 (2007-02-06) * |
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| 袁玉卓等: "《FPGA自学笔记 设计与验证》", 31 August 2017, 北京航空航天大学出版社, pages: 41 * |
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Application publication date: 20221028 |