WO2020038295A1 - Device and method for online heating separation of microbial oil and microbial oil - Google Patents

Abstract

A device and method for online heating separation of microbial oil and the microbial oil. The device comprises: a feed liquid tank (1), a heat exchanger (2), a steam generating device (3), a three-phase centrifuge (4), and a water supply unit (5); the heat exchanger (2) is separately communicated with the feed liquid tank (1), the steam generating device (3), the water supply unit (5), and the three-phase centrifuge (4). The method comprises: exchanging heat between water and steam to allow water to reach a first predetermined temperature, and then allowing water to enter the three-phase centrifuge and to reach a second predetermined temperature; exchanging heat between a lysate and the steam to enable a produced liquid to reach a third predetermined temperature; and allowing the produced liquid to enter the preheated three-phase centrifuge for separation so as to obtain the microbial oil, wherein the lysate is a lysate of a fermentation broth of oleaginous microorganisms.

Description

Device and method for online heating and separating microbial oil and fat, and microbial oil and fat Technical field

The invention relates to the field of separation of microbial oils and fats, in particular to a device for online heating and separation of microbial oils and fats, a method of on-line separation of microbial oils and fats, and microbial oils and fats obtained by the method.

Background technique

Microbial oils and fats are produced by microorganisms such as yeast, mold, bacteria, and algae under certain conditions using carbohydrates, hydrocarbons, and common oils and fats as nitrogen sources, carbon sources, and trace elements for biosynthesis and transformation.

At present, there are two main extraction methods for microbial oil extraction: wet method and dry method. Because of the dry extraction process, the high temperature dehydration leads to large oxidation losses of oil, the need for organic solvent extraction, and high energy consumption. Therefore, more and more microbial oils and fats will be extracted by wet extraction. In the wet process, the primary process task is to break the cell wall using methods such as enzymatic hydrolysis, mechanical disruption, and thermal dissolution. The second is to use the difference in the specific gravity of oil and water to separate the water to obtain the crude oil of microbial oils. . During the separation process, a variety of problems must be solved, such as: the problem of emulsification caused by grease emulsification; the problem of residual oil in the water phase; the problem of residual oil in the slag phase, thereby improving the extraction efficiency.

Conventional demulsification methods include demulsification substances that add electrolytes such as common salt, increase the temperature of the feed solution, and add an acid solution. The addition of electrolytes will cause the production and pollution of hazardous substances such as 3-chloropropanol, and aggravate the production of such pollutants. The method of increasing the temperature of the feed liquid is generally heated in the industry by a fermentation tank or an enzymatic hydrolysis tank. The coil is preheated to about 70 ° C, and then directly heated by steam, and this heating process takes a long time, and it takes more than 10 to heat the tens of tons of enzymolysis solution to the centrifugation process as a whole. Hours, in this way, this long-term heating demulsification method intensifies the oxidation of microbial oils and fats that have been released from the cell wall, resulting in poor quality and great loss of oil; It is usually citric acid or phosphoric acid, which increases the production cost, maintains a certain demulsification time, leads to an increase in peroxide value, and reduces the quality of fats and oils.

In addition, the existing separation methods also have high residual oil content in the water phase and the slag phase after separation, resulting in low extraction efficiency.

Summary of the Invention

The purpose of the present invention is to overcome the above-mentioned problems existing in the prior art, and to provide a microbial oil and fat separation device with low oxidation degree of microbial oils and fats, and low residual oil content in the water phase and slag phase after separation, and high extraction efficiency of oils and fats. A method for separating microbial oils and fats using the device, and a microbial oil and fat obtained therefrom.

In the course of research, the inventor of the present invention found that in the prior art, the following heating methods are generally used to increase the temperature of the material liquid: (1) a holding heating method, which is not uniformly heated, and the cold and hot oils are mixed alternately, which easily leads to Emulsification is easy to occur; (2) When the coil is heated, the grease close to the coil is locally warmed up, and the grease in other areas is lowered by heat transfer. This state of uneven temperature is prone to emulsification when it is stirred. (3) The heating method of spraying high-temperature steam directly into the lysate, the high-temperature steam makes the non-uniformity of the temperature of the grease more obvious, and it is more prone to emulsification; (4) the fermentation tank or the entire fermentation tank needs to be heated, which takes a long time It also accelerates the oxidation of grease.

In order to achieve the above-mentioned object, an aspect of the present invention provides a device for instantaneous heating and separation of microbial oil and fat on-line, the device comprising:

Material liquid tank, heat exchanger, steam supply device, three-phase centrifuge, water supply unit;

Wherein, the heat exchanger is in communication with the material-liquid tank, the steam supply device, the water supply unit and the three-phase centrifuge, respectively;

Wherein, the feed liquid tank is a lysate of an oil-producing microorganism fermentation broth.

Preferably, the heat exchanger is a plate heat exchanger or a tube heat exchanger.

According to a second aspect of the present invention, a method for separating and separating microbial oils and fats online and instantaneously is provided. The method includes:

(1) System preheating: Water is continuously supplied to the heat exchanger by the water supply unit, and steam is continuously supplied to the heat exchanger by the steam supply device to exchange heat between the water flow and the steam in the heat exchanger and make the water The temperature of the effluent water flow is maintained at a first predetermined temperature, and then water is introduced into the three-phase centrifuge to preheat the three-phase centrifuge, so that the three-phase centrifuge reaches a second predetermined temperature;

(2) Material water switching: cut off the water supply of the water supply unit, and continuously supply cracking liquid from the material liquid tank to the heat exchanger, and the steam supply device continues to supply steam to the heat exchanger, and the cracking liquid stream and the steam are made in the heat exchanger. Performing heat exchange and bringing the temperature of the lysate effluent stream to a third predetermined temperature; then introducing the lysate into a preheated three-phase centrifuge for separation of the oil phase, the water phase and the solid phase to obtain microbial oils and fats;

The lysate is a lysate of a fermentation broth of an oil-producing microorganism.

Preferably, the lysate is an enzymatic hydrolysate of a fermentation broth of an oil-producing microorganism, and a method for preparing the enzymatic hydrolysate includes: contacting the fermentation broth of an oleaginous microorganism with a cell wall lyase in a sterile environment to The oil-producing microorganisms in the fermentation broth are subjected to enzymatic hydrolysis to obtain the enzymatic hydrolysis solution.

Preferably, the cell wall lyase includes alkaline protease and optionally other enzymes, and the other enzymes are among cellulase, hemicellulase, pectinase, snail enzyme, chitinase, and ligninase. At least one.

Preferably, the conditions for the enzymolysis include: a pH value of 8-10, a temperature of 40-60 ° C, a pressure of 0.02-0.05MPa, a ventilation volume of 0.2-0.6VVM, and a time of 4-15 hours.

The third aspect of the present invention provides a microbial oil and fat prepared by the method as described above, the microbial oil and fat is a hair oil, and the DHA (docosahexaenoic acid) content of the hair oil is greater than 35% by weight or Tetraenoic acid) content is greater than 35% by weight, 3 chloropropanol <350 μg / kg, anisidine value is less than 25, and peroxide value is less than 20 meq / kg.

In the present invention, only the effluent stream of the lysate is heated by the online instantaneous heating method, so as to meet the temperature requirement of centrifugal separation in the whole process. The thermal efficiency of the heat exchanger is very high, ensuring that the lysate is heated for a short time. And the heating is uniform, avoiding the old-fashioned whole tank of lysate being heated up together and kept under high temperature conditions for a long time, effectively slowing down the rate and degree of oil oxidation, reducing the production cost, and making the final harvested oil quality better than old-fashioned The quality of the fat by centrifugation heating method. The final yield is 98-99%. The quality index of the obtained crude oil is peroxide value ≤20meq / kg, anisidine value ≤25, and the content of DHA and ARA is also increased. In addition, the separation method of the present invention does not need to add substances such as a demulsifier, an electrolyte (for example, 3-chloropropanol), and an acid to the lysate, so that the safety of the product can be more ensured and the cost can be reduced.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a device for separating microbial oil and fat according to a specific embodiment of the present invention.

Reference sign description

1 material liquid tank 2 heat exchanger 3 steam supply device

4-three-phase centrifuge 5 water supply unit 6 first line

7 Second line 8 Third line 9 Temperature monitoring device

10 pressure gauges 11 pressure pumps 12 pressure cups

61First branch line 62 Second branch line

detailed description

The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and these ranges or values should be understood to include values close to these ranges or values. For numerical ranges, between the end values of each range, between the end values of each range and individual point values, and between the individual point values, one or more new numerical ranges can be obtained by combining each other. These values The scope should be considered to be specifically disclosed herein.

In a first aspect, as shown in FIG. 1, the present invention provides a device for on-line separation of microbial oils and fats, the device comprising:

Material liquid tank 1, heat exchanger 2, steam supply device 3, three-phase centrifuge 4, water supply unit 5;

Wherein, the heat exchanger 2 is in communication with the material-liquid tank 1, the steam supply device 3, the water supply unit 5 and the three-phase centrifuge 4 respectively;

Wherein, the feed liquid tank 1 is a lysate of an oil-producing microorganism fermentation broth.

According to the present invention, the connection modes of the material liquid tank 1, the heat exchanger 2, the steam supply device 3, the three-phase centrifuge 4, and the water supply unit 5 are not particularly limited as long as the heat exchanger 2 and the heat exchanger 2 can be ensured respectively. The material-liquid tank 1, the steam supply device 3, the water supply unit 5 and the three-phase centrifuge 4 may communicate with each other. According to a preferred embodiment of the present invention, the heat exchanger 2 is in communication with the material-liquid tank 1 and the water supply unit 5 through a first pipe 6 (that is, both the material-liquid tank 1 and the water-supply unit 5 are in communication with each other). Connected to the branch pipe of the first pipe, preferably, the water supply unit 5 is connected to the first pipe through the first branch pipe 61, and the liquid tank 1 is connected to the first pipe through the second branch pipe 62), The second line 7 is in communication with the steam supply device 3, and the third line 8 is in communication with the three-phase centrifuge 4. Among them, in order to conveniently control the opening and closing of the logistics and the flow rate of the logistics, etc., valves for controlling the logistics are also installed on each pipeline, and the logistics is controlled by the opening and closing of the valves. In this preferred manner, the valve on the first branch pipe 61 and the valve on the second pipe 7 of the water supply unit 5 can be opened first, so that the water provided by the water supply unit 5 and the steam provided by the steam supply device 3 Heat exchange is performed in the heat exchanger 2, after the effluent water reaches a predetermined temperature, the valve on the third pipeline 8 is opened, and the water is introduced into the three-phase centrifuge 4 to preheat it. After the preheating is completed, the The valve on the first branch pipe 61 and the third pipe 8 of the water supply unit 5 are closed, and the valve on the second branch pipe 62 of the material liquid tank 1 is opened, thereby the cracked liquid flows into the heat exchanger 2 After heat exchange with the steam provided by the steam supply device 3, after the output stream of the lysate reaches a predetermined temperature, the valve on the third pipeline 8 is opened again, and the lysate is introduced into the three-phase centrifuge 4 to separate microbial oils and fats. .

According to the present invention, in order to facilitate the switching of material and water, it is preferable that the first branch pipe 61 and the water supply unit 5 are connected through a distribution station, and the second branch pipe 62 and the material liquid tank are also connected through the distribution station. The distribution station distributes water and lysate to the corresponding lines.

According to a preferred embodiment of the present invention, after the feed water is switched, the method further includes draining water in the pipeline through the distribution station, so as not to affect the quality of the lysate. In order to facilitate observing the emptying condition of the water, the distribution station is also preferably connected with a viewing cup 12, and when the color of the liquid flowing through the viewing cup 12 is observed to change, the discharge is stopped. Among them, the discharged water can be directly discharged as sewage or used for other purposes.

According to the present invention, preferably, the water pressure of the water supplied from the water supply unit 5 to the heat exchanger 2 is 0.1-0.2 MPa. The water supply unit 5 may be a water pipe or a water storage container. When the water supply unit 5 is a water pipe, the flow rate of the tap water can be controlled by adjusting the opening of the valve to control the water pressure. When the water supply unit is a water storage container, it can also be provided on the first branch pipe 61. Pressure pump to regulate water pressure.

According to the present invention, in order to further effectively control the heating temperature of the lysate, a pressure pump 11 is further provided on the second branch pipe 62, and the lysate flow is controlled by adjusting the pump pressure of the pressure pump 11 to achieve temperature Precise control. According to a preferred embodiment of the present invention, the magnitude of the pump pressure is preferably such that the pressure of the cracked liquid stream entering the heat exchanger 2 is 0.1-0.2 MPa. The pressure pump 11 may be a rotor pump.

Therefore, preferably, a pressure gauge 10 is further provided on the first pipeline 6.

According to a preferred embodiment of the present invention, in order to improve the heat exchange efficiency, the heat exchanger 2 is a plate heat exchanger or a tube heat exchanger.

According to a preferred embodiment of the present invention, in order to facilitate the monitoring of the temperature of the stream entering the three-phase centrifuge 4, a temperature monitoring device is also provided on the pipeline where the heat exchanger 2 communicates with the three-phase centrifuge 4. 9, wherein the temperature monitoring device 9 is disposed near one end of the heat exchanger 2, and is more preferably disposed at a logistics outlet of the heat exchanger 2. The temperature monitoring device 9 may be a thermometer.

According to the present invention, the feed liquid tank 1 may be any feed liquid tank containing a lysate of an oil-producing microorganism fermentation liquid, for example, it may be an enzymatic hydrolysis tank or a fermentation tank.

In a second aspect, the present invention provides a separation method for instantaneous heating and separation of microbial oils and fats, which is characterized in that the method includes:

(1) System preheating: Water is continuously supplied to the heat exchanger by the water supply unit, and steam is continuously supplied to the heat exchanger by the steam supply device to exchange heat between the water flow and the steam in the heat exchanger and make the water The temperature of the effluent water flow is maintained at a first predetermined temperature, and then water is introduced into the three-phase centrifuge to preheat the three-phase centrifuge, so that the three-phase centrifuge reaches a second predetermined temperature;

(2) Material water switching: cut off the water supply of the water supply unit, and continuously supply cracking liquid from the material liquid tank to the heat exchanger, and the steam supply device continues to supply steam to the heat exchanger, and the cracking liquid stream and the steam are made in the heat exchanger. Performing heat exchange and bringing the temperature of the lysate effluent stream to a third predetermined temperature; then introducing the lysate into a preheated three-phase centrifuge for separation of the oil phase, the water phase and the solid phase to obtain microbial oils and fats;

The lysate is a lysate of a fermentation broth of an oil-producing microorganism.

According to the present invention, the method for performing on-line separation of microbial oils and fats is performed in an on-line microbial oil and fat separation device as described above. The invention enters the heat exchanger in the form of an enzymatic hydrolysate stream and instantly heats it to a predetermined temperature online, thereby avoiding heating the entire tank of the enzymatic hydrolysate, thereby causing the enzymatic hydrolysate to be exposed to high temperature for a long time, and the In the form of side-by-side flow heating, the heating can be more uniform. In addition, heating of demulsifiers, electrolytes, acids, etc. is saved, and the resulting crude oil is not polluted. In order to further improve the heating efficiency, the heat exchanger is a plate heat exchanger or a tube heat exchanger.

Preferably, the first predetermined temperature may be 80-95 ° C.

Preferably, the second predetermined temperature may be 80-95 ° C.

Preferably, the third predetermined temperature may be 80-95 ° C.

According to the present invention, the water supply unit continuously supplies water to the heat exchanger by transmitting water to the distribution station and passing through the first branch pipe 61 and the first pipe 6; the water supply pressure is preferably 0.1-0.2 MPa;

According to the present invention, the lysate from the feed liquid tank enters the replacement tap water from the distribution station, and continuously supplies the lysate to the heat exchanger through the second branch pipe 62; The hydraulic pressure is preferably 0.1-0.2MPa;

According to the present invention, preferably, after the feed water is switched, the method further includes discharging water in the distribution station, the first branch pipe 61 and the first pipe 6. Because the color of the water and the material liquid is quite different, you can observe whether the color change water is exhausted through the viewing cup 12 provided at the sewage pipe of the distribution station.

According to the present invention, after the feed water is switched, the method further includes discharging hot water for preheating in the three-phase centrifuge.

According to the present invention, the centrifugation time may be 15-20 hours.

According to the present invention, in order to ensure the effective performance of heat exchange, before the start of the operation of the device, it also includes a water leakage check on the heat exchanger. For example, water can be supplied to the heat exchanger through a water supply unit to check whether the heat exchanger leaks water.

According to the present invention, preferably, when the pre-heated water in step (1) enters the three-phase centrifuge, the three-phase centrifuge is in a full-speed starting state, and its rotation speed may be 5000-8000 rpm.

According to the present invention, preferably, when the pre-heated lysate in step (2) enters the three-phase centrifuge, the three-phase centrifuge is in a full-speed starting state, and its rotation speed may be 5000-8000 rpm, that is, the separated The speed is 5000-8000rpm.

According to the present invention, the lysate may be a lysate obtained by breaking the microorganisms in the oil-producing microorganism fermentation broth by various methods conventional in the prior art, for example, by lysing, enzymatic hydrolysis, mechanical crushing, thermal dissolution and other methods. Wall products.

When the wall was broken by enzymatic hydrolysis, the inventors of the present invention found during the research that the main reasons for the low extraction efficiency of microbial oils and fats, the high content of anisidine and peroxide values, and the low content of DHA or ARA Another aspect is that in the current process, substances such as enzyme preparations and pH adjusters need to be introduced into the fermentation concentrate, and the enzyme preparations and pH adjusters are usually added directly to the concentrated fermentation broth after the preparation, which will result in Contamination of fermentation broth by foreign bacteria.

Generally, the entire process takes more than 20 hours from the adjustment of pH, the preparation and dosing of enzyme preparations, the implementation of enzymolysis, to the post-heating treatment. Bacterial microorganisms multiply very quickly. Generally, one generation occurs every 20 minutes. During the entire process of extracting fat, over 20 hours, the amount of bacteria in the fermentation broth rich in carbon and nitrogen is very large. At the same time, although the enzymatic hydrolysis at the later stage can kill a certain amount of microorganisms, microorganisms such as bacteria will secrete a large amount of exotoxin in the process. Thermal heating can only kill living organisms of bacterial microorganisms, and exotoxin is a type Protein, heat has a limited effect on it, and spore exotoxin enters the fat as a protein fragment or peptide, which becomes a factor affecting the safety of the fat. In addition, due to the presence of a large number of foreign bacteria, it will compete with oil-producing microorganisms during the treatment process, thereby weakening the treatment effect on oil-producing microorganisms, causing a large amount of foreign bacteria's intracellular substances to enter the obtained microbial oils, resulting in The extraction efficiency of microbial oils and fats is low, and the content of main components such as DHA and ARA is reduced. In addition, because of bacterial infection and large-scale reproduction, the fermentation broth produces a malodor during the treatment process, which affects the production environment.

Therefore, preferably, the lysate is an enzymatic hydrolysate of a fermentation broth of an oil-producing microorganism, and a method for preparing the enzymatic hydrolysate includes: contacting a fermentation broth of an oil-producing microorganism with a cell wall lyase in a sterile environment, Enzymolysis is performed on the oil-producing microorganisms in the fermentation broth to obtain the enzyme hydrolysate.

In the present invention, it should be noted that the sterile environment means that the enzymatic system is in a sterile environment except for the oil-producing microorganisms contained in the fermentation broth.

According to the present invention, the fermentation broth of the oil-producing microorganism is a fermentation broth obtained directly after fermentation of the oil-producing microorganism, and does not need to undergo any further treatment. Therefore, in the above preferred case, compared with the prior art, the present application The method also saves the concentration of fermentation broth and the inactivation steps of oil-producing microorganisms.

According to the present invention, a method for preparing a fermentation broth of an oil-producing microorganism is well known to those skilled in the art, for example, inoculating an oil-producing microorganism into a fermentation sugar solution for fermentation, thereby obtaining a fermentation broth of an oil-producing microorganism.

The oil-producing microorganisms can be various existing oil-producing microorganisms. For example, the oil-producing microorganisms can be any one of bacteria, molds, yeasts, and algae. Preferably, the oil-producing microorganisms are molds, yeasts, and algae. Any of them. Examples of the molds may include, but are not limited to, Asoergullus terreus, Clavicepspurpurea, Tolyposporium, Mortierella alpina, and Mortierella Mortierella isabibellina; examples of the yeast may include, but are not limited to, Cryptococcus albidus, Cryptococcus albidun, Lipomyces, and Mycelium Trichospiron pullulans, Lipomy slipperer, Rhodotorula giutinis and Rhodosporidium tortorides; examples of the algae may include, but are not limited to, thraustochytrids (Thraustochytriales), Schizochytrium, Crypthecodinium, diatom, Spirulina, and Wokenella.

It is well known to those skilled in the art that (1) the extracellular matrix of an animal is a cell wall in a sense, and its chemical composition is collagen, adhesion protein, amino polysaccharide, and proteoglycan. (2) The main component of bacterial cell wall is peptidoglycan. (3) The main components in the fungal cell wall are chitin, cellulose, dextran, mannan, etc. These polysaccharides are all polymers of monosaccharides. (4) The plant cell wall is mainly cellulose, hemicellulose and pectin, and there is also a large amount of lignin in the secondary cell wall. Generally, the oil-producing microorganisms are bacteria, fungi, yeast, and algae. Therefore, the enzymatic hydrolysis of the cell walls of oil-producing microorganisms is usually limited to cellulase, hemicellulase, pectinase, snail enzyme, and chitin. Enzymes, ligninase, etc. However, in the process of cracking the cell wall of oil-producing microorganisms by the currently used enzyme preparations, the breaking performance of the enzyme preparations is not stable. Whether liquid or solid enzymes have large fluctuations in performance, which makes it difficult to grasp the production operation. The inventor of the present invention unexpectedly discovered during the research that by introducing an alkaline protease into the enzyme preparation, not only the excellent enzymatic hydrolysis efficiency can be ensured, but also the enzymatic hydrolysis process can be performed stably, thereby further improving the extraction efficiency of microbial oils and fats. Decrease anisidine value and peroxide value, increase DHA or ARA content.

Therefore, according to a preferred embodiment of the present invention, the cell wall lyase comprises an alkaline protease. In addition, preferably, the cell wall lyase may further include at least one of a cellulase, a hemicellulase, a pectinase, a snail enzyme, a chitinase, and a ligninase.

According to the present invention, the amount of the cell wall lyase can be selected in a wide range, as long as the cell wall of the oil-producing microorganism can be fully lysed, thereby releasing the microbial oil and fat. Preferably, the amount of the cell wall lyase is 1-5 g per liter of the fermentation broth.

According to the present invention, the conditions for the enzymolysis may be those conventionally used for lysing the cell wall of an oil-producing microorganism by enzymatic hydrolysis, but the inventors of the present invention have found that by performing the enzymolysis under aerobic conditions, The enzymolysis efficiency is further improved, thereby improving the extraction efficiency of microbial oils and fats, and the DHA or ARA of the obtained microbial oils and fats. Therefore, preferably, the conditions for the enzymolysis include: a pH value of 8-10 (for example, it can be 8, 8.5, 9, 9.5, and 10), and a temperature of 40-60 ° C (for example, it can be 40 ° C, 45 ℃, 50 ℃, 55 ℃, 60 ℃), the pressure is 0.02-0.05MPa (for example, it can be 0.02MPa, 0.03MPa, 0.04MPa, 0.05MPa), and the ventilation volume is 0.2-0.6VVM (per unit volume per minute per minute The amount of gas passed through the fermentation broth is 0.2-0.6 volume) (for example, it can be 0.2VVM, 0.3VVM, 0.4VVM, 0.5VVM, 0.6VVM), and the time is 4-15 hours (for example, it can be 4 hours, 5 hours) , 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours).

According to the present invention, in order to further improve the contact between the enzyme and the oil-producing microorganisms, and thereby the enzymatic hydrolysis efficiency, the enzymatic hydrolysis is performed under the condition of stirring, and the stirring speed may be 8-30 rpm.

Among them, the pH value of the enzymatic hydrolysis system can be controlled to 8-10 by adding lye to the fermentation broth. In the present invention, the selection of the lye is not particularly limited, as long as it can complete the adjustment of the pH and does not cause side reactions with the generated microbial oils and fats. Examples of the lye may include, but are not limited to, at least one of a sodium oxide solution, a potassium hydroxide solution, a sodium carbonate solution, a sodium bicarbonate solution, a potassium carbonate solution, a potassium bicarbonate solution, and ammonia water. The concentration of the lye is not particularly limited in the present invention, as long as the pH can be adjusted. For example, the concentration of the lye can be 15-25% by weight.

Preferably, in order to ensure the safety of the finally obtained microbial oil and fat, the lye is a food-grade lye.

According to the present invention, the enzymolysis can be carried out directly in a fermentation tank. In order to ensure the aseptic environment of the system, the lye is preferably transported to the fermentation tank through a sterile pipeline after being sterilized, and the cell wall is lysed. The enzyme is formulated into a solution with the above enzyme activity, and after being sterilized, the enzyme is delivered to the fermentation tank through a sterile pipeline.

Among them, the method for sterilizing the lye and the pipeline for transmitting the lye and cell wall lyase can be various sterilization methods known in the art, for example, a filtration method, an ozone disinfection method, and a high temperature saturation method. Steam method. In the present invention, the lye and the pipeline for delivering the lye and cell wall lyase are preferably sterilized by a method of high temperature saturated steam. The pressure of the high-temperature saturated steam may be 0.1-0.35 MPa, the temperature may be 121-145 ° C, and the sterilization time may be 40-80 min.

Among them, in order to ensure the activity of the cell wall lyase, the conventional method of aseptic filtration of the present invention can be adopted for the method of the cell wall lyase, for example, the bacteria can be sterilized by using a filtration system for filtering the sterile liquid of the filtration system. The aperture of the device can be 0.15-0.25 μm. Preferably, the filtering system further includes a step of sterilizing the filter system before use. The sterilization method may be a method of ozone disinfection, a method of high temperature saturated steam, and other methods known in the art. In the present invention, the filtering system is preferably sterilized by a method of high temperature saturated steam. The pressure of the high-temperature saturated steam may be 0.1-0.35 MPa, the temperature may be 121-145 ° C, and the sterilization time may be 40-80 min.

According to the present invention, the enzymolysis can also be performed in an enzymolysis tank. In order to ensure the aseptic environment of the system, the lye is preferably transported to the enzymolysis tank through a sterile pipeline after being sterilized. The cell wall lysing enzyme is formulated into the above enzyme activity solution, sterilized by a sterile liquid filter, and then transferred to the enzymatic hydrolysis tank through a sterile pipeline, and the fermentation broth is transferred to the enzymatic hydrolysis tank through a sterile pipeline. in.

The method for sterilizing the lye and the pipeline for conveying the lye, the cell wall lyase and the fermentation broth may be various sterilization methods known in the art, for example, a filtration method, an ozone disinfection method , Method of high temperature saturated steam. In the present invention, the lye and the pipeline for conveying the lye, the cell wall lyase and the fermentation broth are preferably sterilized by a method of high temperature saturated steam. The pressure of the high-temperature saturated steam may be 0.1-0.35 MPa, the temperature may be 121-145 ° C, and the sterilization time may be 40-80 min.

Among them, in order to ensure the activity of the cell wall lyase, the conventional method of sterilization at normal temperature of the present invention can be used for the method of the cell wall lyase, for example, the bacteria can be sterilized by using a filtration system, which is a liquid sterile filter. The pore size can be 0.15-0.25 μm. Preferably, the filtering system further includes a step of sterilizing the filter system before use. The sterilization method may be a method of ozone disinfection, a method of high temperature saturated steam, and other methods known in the art. In the present invention, the filtering system is preferably sterilized by a method of high temperature saturated steam. The pressure of the high-temperature saturated steam may be 0.1-0.35 MPa, the temperature may be 121-145 ° C, and the sterilization time may be 40-80 min.

A specific embodiment of the present invention will now be described in detail with reference to FIG. 1. Specifically,

(1) Preparation of lye: add a certain amount of water to the alkali tank, weigh NaOH at a concentration of 15-25% by weight, and dissolve it in the alkali tank. Put the prepared lye in high-temperature saturated steam for 1 h, then Pass the sterile compressed air into the alkali tank to make the tank pressure of the alkali tank be 0.02-0.05 MPa, and pass cooling water into the jacket of the alkali tank to cool the alkali solution to 35-45 ° C, and reserve it.

(2) Preparation of cell wall lysate: Mix the cell wall lysate with a certain amount of water and prepare it in the ratio of enzyme: water = 1: (5-10). Sterilize the filtration system with high temperature saturated steam for 1 h, then use Sterile compressed air is kept under pressure.

(3) Dosing of the enzymatic hydrolysis solution and the lye: first raise the temperature of the fermentation liquid to 35-55 ° C, and then hydraulically press the alkali into the fermentation tank through a sterile pipeline to make the pH of the fermentation liquid 8-10; The above-mentioned enzymolysis solution is transferred into a sterile filtration system through a diaphragm pump and filtered, and then transported into a fermentation tank through a sterile pipeline to perform enzymolysis of the fermentation solution. The entire enzymolysis time is 4-15h.

(4) Preheating preparation of plate heat exchanger: First, water is supplied to plate heat exchanger 2 from water supply unit 5 to ensure that the plates of plate heat exchanger 2 are well sealed, and then steam supply device 3 is used to plate heat exchanger. Slowly inject steam and water for heat exchange, so that the outlet temperature of heat exchanger 2 slowly rises, and then gradually increase the supply of steam to ensure that the outlet temperature is 80-95 ° C after heat exchange;

(5) Preheating preparation of the centrifuge: Pass hot water of 80-95 ° C into the three-phase centrifuge 4 so that the temperature of the centrifuge drum body reaches 80-95 ° C;

(6) Feed centrifugation: The water in the feed pipe of the plate heat exchanger 2 is switched to the hydrolyzed enzymatic solution. When the temperature of the hydrolyzed solution reaches 80-95 ° C, the hydrolyzed solution is delivered to The three-phase centrifuge 4 starts the centrifugation process during the centrifugation process.

In a second aspect, the present invention provides a microbial oil and fat prepared by the method described above, the microbial oil and fat is hair oil, and the DHA content of the hair oil is greater than 35% by weight or the ARA content is greater than 35% by weight, 3 chlorine Propanol is less than 350 μg / kg, anisidine value is less than 25, and peroxide value is less than 20 meq / kg.

Hereinafter, the present invention will be described in detail through examples. In the following embodiments,

Alkaline protease was purchased from Danisco, PD 216661-7.0CHN;

Pectinase was purchased from Dongheng Huadao Biotechnology Co., Ltd., P128776;

Cellulase was purchased from Jiangsu Yihaotian Biotechnology Co., Ltd., article number 232-734-4;

The oil-producing microbial fermentation broth 1 is a fermented broth obtained by fermenting mildew in high mountains, mainly containing ARA;

The oil-producing microorganism fermentation broth 2 is a fermentation broth obtained by Schizochytrium fermentation, which mainly contains DHA;

The DHA content in the obtained crude oil was determined by gas chromatography according to the method of GB26400-2011;

EPA content was determined by gas chromatography in accordance with the method of GB5009.168-2016;

ARA content was measured by gas chromatography according to GB26401-2011 method;

3-chloropropanol content was determined by high performance liquid chromatography;

Anisidine value was measured by GB / T 24304-2009 method;

Peroxide value is measured by ultraviolet spectrophotometer according to GB / T24304-2009 method;

The yield of microbial oils and fats is measured by a method of extraction and weighing with a rotary evaporator;

The residual oil ratio in the solid residue was measured by a rotary evaporator extraction weighing method.

The device for separating microbial oils as shown in Fig. 1 includes: material liquid tank 1 (fermentation tank), heat exchanger 2 (plate heat exchanger), steam supply device 3, three-phase centrifuge 4 (purchased from Yixinghua, Jiangsu) Dingliang Oil Machinery Co., Ltd., article number BTSD95), water supply unit 5 (tap water pipe); the heat exchanger 2 communicates with the liquid-liquid tank 1 and the water supply unit 5 through the first pipeline 6, and communicates with the The steam supply device 3 is in communication, and is in communication with the three-phase centrifuge 4 through a third pipe 8; a return pipe 9 is further provided between the material-liquid tank 1 and the heat exchanger 2; a heat exchanger A temperature monitoring device 10 is provided at the discharge port of 2. The water supply unit 5 is connected to the first pipeline 6 through a distribution station and a first branch pipeline 61; the material liquid tank 1 is connected to the first pipeline (6) through the distribution station and a second branch pipeline 62 A pressure pump 11 is provided on the second branch pipe 62; a pressure gauge 10 is provided on the first pipe 6; and a viewing cup 12 is also connected to the distribution station.

Example 1

This embodiment is used to explain the microbial oil and fat provided by the present invention and a preparation method thereof

(1) Preparation of lye: Add 200L of demineralized water to the alkali tank, weigh out food-grade NaOH at a concentration of 20% by weight, and dissolve it in the alkali tank. Place the prepared lye at a pressure of 0.14MPa and temperature Sterilize in saturated steam at 145 ° C for 1 hour, then pass sterile compressed air into the alkali tank to make the tank pressure of the alkali tank be 0.02-0.05MPa, and pass cooling water into the jacket of the alkali tank to cool the alkali solution to 35-45 ℃, spare;

(2) Preparation of cell wall lysate: sterilize the secondary liquid sterile filtration system in series with a pressure of 0.14 MPa and a temperature of 145 ° C and sterilize it with saturated steam for 1 hour, and then maintain the pressure with sterile compressed air for standby; the sterile filtration The filter specifications of the system are 0.2 μm in pore size and 150 ° C temperature resistance. Weigh alkaline protease and add demineralized water to prepare an enzymatic hydrolysate at an enzyme: water ratio of 1:10.

(3) Dosing of enzymatic hydrolysis liquid and alkaline liquid: first raise the temperature of the oil-producing microorganism fermentation liquid 1 to 35-55 ° C, and then press the above alkaline liquid into the fermentation tank with compressed air through a sterile alkaline liquid pipeline, The pH of the fermentation broth was measured with a "METTLER" pH probe, and the lye was stopped from being added until the pH of the fermentation broth was 9; the above-mentioned enzymatic hydrolysate was transferred to a serially connected secondary liquid sterile filtration system through a diaphragm pump, and then filtered The aseptic pipeline is transported into the fermentation tank, and the amount of enzyme added is 3 g per liter of fermentation broth, and the enzymatic hydrolysis of the fermentation broth is performed; during the enzymatic hydrolysis, the temperature is controlled to 55 ° C, the pH is controlled to 9, and the stirring speed is controlled to At 20 rpm, the ventilation volume is controlled to 0.45 vvm, the tank pressure is controlled to 0.03 MPa, and the entire enzymolysis time is 8 h.

(4) Preheating preparation for plate heat exchanger 5: Open the valve between the water pipe and the plate heat exchanger, and the water pipe supplies water to the plate heat exchanger through the distribution station. The water supply pressure is 0.1-0.2MPa. The plate seal of the heat exchanger is determined. After confirming that the plate seal is in good condition, slowly open the valve between the plate heat exchanger and the steam supply device, and provide steam to the plate heat exchanger through the steam supply device to exchange heat with the water in the plate heat exchanger. The temperature of the water was detected by the temperature monitoring device, and it was determined that it reached 90 ° C;

(5) Pre-heating preparation of three-phase centrifuge: Turn on the cooling water of the three-phase centrifuge to ensure the smooth flow of water at the outlet. At the same time, turn on the three-phase centrifuge 13 to ensure that the starting current is 80A and the full-speed indicator plate speed is 6700rpm. The valve between the heat exchanger and the three-phase centrifuge allows 90 ° C hot water to enter the three-phase centrifuge, so that the temperature of the drum body of the three-phase centrifuge reaches 90 ° C;

(6) Feed centrifugation: Open the valve between the fermentation tank and the plate heat exchanger, and close the valve between the plate heat exchanger and the water pipe at the same time, and switch the tap water in the first pipe to step (4) through the distribution station ), And open the drain valve of the distribution station to drain the tap water in the pipeline. When the color change is observed in the sight cup, the sewage is stopped, and the temperature of the digested solution entering the heat exchanger for heat exchange reaches 90. At ℃, the enzymolysis solution reaching 90 ℃ was transferred to a three-phase centrifuge and centrifuged for 18 hours to obtain microbial oils and fats as crude oil, extraction efficiency of oils and fats, residual oil rate in solid phase residues, ARA content in oils and fats, EPA The content, 3 chloropropanol content, anisidine value, and peroxide value are shown in Table 1.

Example 2

This embodiment is used to explain the microbial oil and fat provided by the present invention and a preparation method thereof

(1) Preparation of lye: Add 200L of demineralized water to the alkali tank, weigh food-grade sodium carbonate at a concentration of 15% by weight, and dissolve it in the alkali tank. Place the prepared lye at a pressure of 0.14MPa. Sterilize in saturated steam at 145 ° C for 1 hour, then pass sterile compressed air into the alkali tank to make the tank pressure of 0.02-0.05MPa, and pass cooling water into the jacket of the alkali tank to cool the alkali solution. To 35-45 ℃, spare;

(2) Preparation of cell wall lysate: sterilize the secondary liquid sterile filtration system in series with a pressure of 0.14 MPa and a temperature of 145 ° C and sterilize it with saturated steam for 1 hour, and then maintain the pressure with sterile compressed air for standby; the sterile filtration The filter specifications of the system are 0.2 μm in pore size and 150 ° C temperature resistance. Weigh alkaline protease, pectinase, cellulase, and snail enzyme, and add demineralized water to prepare an enzymatic hydrolysate at an enzyme: water ratio of 1:10. The amount of enzyme per liter of fermentation broth is: 2g alkaline protease, 1g pectinase, 1.5g cellulase, and 0.5g snail enzyme.

(3) Dosing of enzymatic hydrolysis liquid and alkaline liquid: first raise the temperature of the oil-producing microorganism fermentation liquid 1 to 35-55 ° C, and then press the above alkaline liquid into the fermentation tank with compressed air through a sterile alkaline liquid pipeline, The pH of the fermentation broth was measured with a "METTLER" pH probe, and the lye was stopped from being added until the pH of the fermentation broth was 8; the above-mentioned enzymolysis solution was transferred to a serially connected secondary liquid sterile filtration system through a diaphragm pump, and then filtered through The aseptic pipeline is transported into the fermentation tank for enzymatic hydrolysis of the fermentation broth; during the enzymatic hydrolysis, the temperature is controlled at 50 ° C, the pH is controlled at 8, the stirring speed is controlled at 30 rpm, the ventilation volume is controlled at 0.2 vvm, and the tank pressure is controlled at 0.02 MPa, the entire enzymolysis time is 15h.

(4) Preheating preparation for plate heat exchanger 5: Open the valve between the water pipe and the plate heat exchanger, and the water pipe supplies water to the plate heat exchanger through the distribution station. The water supply pressure is 0.1-0.2MPa. The plate seal of the heat exchanger is determined. After confirming that the plate seal is in good condition, slowly open the valve between the plate heat exchanger and the steam supply device, and provide steam to the plate heat exchanger through the steam supply device to exchange heat with the water in the plate heat exchanger. The temperature of the water was detected by the temperature monitoring device, and it was determined that it reached 95 ° C;

(5) Preheating preparation for three-phase centrifuge: Turn on the cooling water of the three-phase centrifuge to ensure smooth water flow at the water outlet. At the same time, turn on the three-phase centrifuge 13 to ensure that the starting current is 80A and the full-speed indicator plate speed is 6000 rpm. The valve between the heat exchanger and the three-phase centrifuge allows 95 ° C hot water to enter the three-phase centrifuge, so that the temperature of the drum body of the three-phase centrifuge reaches 95 ° C;

(6) Feed centrifugation: Open the valve between the fermentation tank and the plate heat exchanger, and close the valve between the plate heat exchanger and the water pipe at the same time, and switch the tap water in the first pipe to step (4) through the distribution station ) The obtained enzymolysis solution, and open the drain valve of the distribution station to drain the tap water in the pipeline. When the color change is observed in the sight cup, the drainage is stopped, and the temperature of the enzymolysis solution entering the heat exchanger for heat exchange reaches 95 At ℃, the reflux of the enzymolysis solution was stopped, and the enzymolysis solution that reached 95 ° C was transferred to a three-phase centrifuge, and centrifuged for 15 hours to obtain microbial oils and fats as crude oil, the extraction efficiency of oils and fats, and the residual oil ratio in solid residues. See Table 1 for ARA content, EPA content, 3chloropropanol content, anisidine value, and peroxide value in fats and oils.

Example 3

This embodiment is used to explain the microbial oil and fat provided by the present invention and a preparation method thereof

(1) Preparation of lye: Add 200L of demineralized water to the alkali tank, weigh out food-grade sodium bicarbonate at a concentration of 25% by weight, and dissolve it in the alkali tank. Place the prepared lye at a pressure of 0.14MPa , Sterilize in saturated steam at 145 ℃ for 1h, then pass sterile compressed air into the alkali tank to make the tank pressure of the alkali tank be 0.02-0.05MPa, and pass cooling water into the jacket of the alkali tank to lye Cool to 35-45 ° C and reserve;

(2) Preparation of cell wall lysate: sterilize the secondary liquid sterile filtration system in series with a pressure of 0.14 MPa and a temperature of 145 ° C and sterilize it with saturated steam for 1 hour, and then maintain the pressure with sterile compressed air for standby; the sterile filtration The filter specifications of the system are 0.2 μm in pore size and 150 ° C temperature resistance. Weigh alkaline protease and add demineralized water to prepare the enzymatic hydrolysis solution according to enzyme: water = 1: 10.

(3) Dosing of enzymatic hydrolysis liquid and alkaline liquid: first raise the temperature of the oil-producing microorganism fermentation liquid 1 to 35-55 ° C, and then press the above alkaline liquid into the fermentation tank with compressed air through a sterile alkaline liquid pipeline, Use the "METTLER" pH probe to measure the pH of the fermentation broth until the pH of the fermentation broth reaches 10; stop adding lye; the above-mentioned enzymolysis solution is pumped through a diaphragm pump into a serially connected secondary liquid sterile filtration system, and then filtered through The aseptic pipeline is transported into the fermentation tank, and the amount of enzyme added is 5 g per liter of fermentation broth to perform the enzymatic hydrolysis of the fermentation broth; during the enzymatic hydrolysis, the temperature is controlled to 50 ° C, the pH is controlled to 10, and the stirring speed is controlled to At 8 rpm, the ventilation volume is controlled to 0.6 vvm, the tank pressure is controlled to 0.05 MPa, and the entire enzymatic hydrolysis time is 4 h.

(4) Preheating preparation for plate heat exchanger 5: Open the valve between the water pipe and the plate heat exchanger, and the water pipe supplies water to the plate heat exchanger through the distribution station. The water supply pressure is 0.1-0.2MPa. The plate seal of the heat exchanger is determined. After confirming that the plate seal is in good condition, slowly open the valve between the plate heat exchanger and the steam supply device, and provide steam to the plate heat exchanger through the steam supply device to exchange heat with the water in the plate heat exchanger. The temperature of the water was detected by the temperature monitoring device, and it was determined that it reached 85 ° C;

(5) Preheating preparation for three-phase centrifuge: Turn on the cooling water of the three-phase centrifuge to ensure smooth water flow at the water outlet. At the same time, turn on the three-phase centrifuge 13 to ensure that the starting current is 80A and the full-speed indicator plate speed is 7500rpm. The valve between the heat exchanger and the three-phase centrifuge allows hot water at 85 ° C to enter the three-phase centrifuge, so that the temperature of the drum body of the three-phase centrifuge reaches 85 ° C;

(6) Feed centrifugation: Open the valve between the fermentation tank and the plate heat exchanger, and close the valve between the plate heat exchanger and the water pipe at the same time, and switch the tap water in the first pipe to step (4) through the distribution station ), And open the drain valve of the distribution station to drain the tap water in the pipeline. When the color change is observed in the sight cup, the sewage is stopped, and the temperature of the digested solution entering the heat exchanger for heat exchange reaches 85. At ℃, the reflux of the enzymolysis solution was stopped, and the enzymolysis solution that reached 85 ° C was transferred to a three-phase centrifuge and centrifuged for 20 hours to obtain microbial oils and fats as the crude oil, the extraction efficiency of the oils and oil, and the residual oil rate in the solid phase residue. See Table 1 for ARA content, EPA content, 3chloropropanol content, anisidine value, and peroxide value in fats and oils.

Example 4

The microbial oil and fat preparation was performed according to the method of Example 1, except that the oil-producing microorganism fermentation broth 1 in step 3 was replaced with the oil-producing microorganism fermentation broth 2. See Table 1 for the extraction efficiency of oils and fats, the residual oil rate in solid phase residues, the DHA content in oils and fats, the EPA content, the 3 chloropropanol content, the anisidine value, and the peroxide value.

Example 5

The microbial oil and fat preparation was performed according to the method of Example 2, except that the oil-producing microbial fermentation broth 1 in step 3 was replaced with the oil-producing microbial fermentation broth 2. See Table 1 for the extraction efficiency of oils and fats, the residual oil rate in solid phase residues, the DHA content in oils and fats, the EPA content, the 3 chloropropanol content, the anisidine value, and the peroxide value.

Example 6

The microbial oil and fat preparation was carried out according to the method of Example 3, except that the oil-producing microorganism fermentation broth 1 in step 3 was replaced with the oil-producing microorganism fermentation broth 2. See Table 1 for the extraction efficiency of oils and fats, the residual oil rate in solid phase residues, the DHA content in oils and fats, the EPA content, the 3 chloropropanol content, the anisidine value, and the peroxide value.

Example 7

This embodiment is used to explain the microbial oil and fat provided by the present invention and a preparation method thereof

The microbial oil was prepared according to the method of Example 1. The difference was that the enzyme used for enzymolysis was replaced with snail enzyme, cellulase and pectinase. The amount of enzyme used per liter of fermentation broth was 2.5g of cellulase. The results are shown in Table 1 with 1.5 g of pectinase and 1 g of snail enzyme.

Example 8

This comparative example is used to explain the reference microbial oil and fat and its preparation method

The preparation of microbial oils and fats was performed according to the method of Example 1. The difference is that the lye, the enzymatic hydrolysis solution, and the pipelines that transport the lye and the enzymatic hydrolysis solution did not pass the sterilization. The results are shown in Table 1.

Example 9

This comparative example is used to explain the reference microbial oil and fat and its preparation method

The microbial oil and fat preparation was performed according to the method of Example 4, except that the lye, the enzymatic hydrolysis solution, and the pipelines for conveying the lye and the enzymatic hydrolysis solution did not pass the sterilization. The results are shown in Table 1.

Comparative Example 1

The preparation of microbial oils and fats was performed according to the method of Example 1, except that the entire enzymolysis solution was heated to 90 ° C. in a fermentation tank and then centrifuged. The results are shown in Table 1.

Comparative Example 2

This comparative example is used to explain the reference microbial oil and fat and its preparation method

The preparation of microbial oil and fat was performed according to the method of Example 4, except that the entire enzymolysis solution was heated to 90 ° C. in the fermentation tank and then centrifuged. The results are shown in Table 1.

Table 1

It can be seen from Table 1 that the method for separating microbial oils and fats of the present invention does not add any demulsifier, has high safety, and the yield of the microbial oils and fats obtained as crude oil can be increased to 98-99%, compared with the control. In addition, the DHA content or ARA content in the hair oil was significantly increased, and the trichloropropanol content, anisidine value, and peroxide value were significantly reduced; in addition, the entire process was controlled to Operating in a bacterial environment can reduce the proliferation of bacteria during the extraction of microbial oils, further increase the safety of microbial oils, the yield of microbial oils, the DHA content or ARA content in hair oil, and further reduce the anisidine value and peroxides value. In addition, the effect can be further improved under the preferred use of alkaline protease and the preferred enzymatic hydrolysis conditions.

Test case

The degree of enzymatic hydrolysis is usually measured by the following two methods:

(a) Microscope to observe the outline morphology of algal cells: After the end of the enzymolysis, take a sample of 200mL from the sampling port of the fermentation tank with an Erlenmeyer flask, pick out 2-4 loops of the enzymatic fermentation broth with the inoculating ring, and spread it on the coverslip and cover Observe the slide, drop 1-2 drops of cedar oil, and observe with a 100x oil lens. See if there are complete algae cells; whether there are large pieces of cell debris, the results are shown in Table 2.

(b) Centrifugal observation method: After the end of the enzymolysis, take a sample of 200mL in a conical flask at the sampling port of the fermenter, take 50mL of the fermentation broth with a 50mL centrifuge tube, and centrifuge in a laboratory centrifuge at 6000-10000rpm for 3min, observe the oil Phase and water phase separation, to see if the layered interface is clear. Use a 1-10 point system to score. The higher the score, the clearer. The results are shown in Table 2.

Table 2

It can be seen from Table 2 that under the preferred use of alkaline protease and the preferred enzymatic hydrolysis conditions, the effect can be further improved.

The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the scope of the technical idea of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, including the combination of various technical features in any other suitable manner. These simple modifications and combinations should also be regarded as the disclosure of the present invention. All belong to the protection scope of the present invention.

Claims (10)

1. A device for instantaneously heating and separating microbial oil and fat on-line, which is characterized in that the device includes:

   Material liquid tank (1), heat exchanger (2), steam supply device (3), three-phase centrifuge (4), water supply unit (5);

   The heat exchanger (2) is in communication with the material-liquid tank (1), the steam supply device (3), the water supply unit (5), and the three-phase centrifuge (4), respectively;

   Wherein, the feed liquid tank (1) is a lysate of an oil-producing microorganism fermentation broth.
2. The device according to claim 1, wherein the heat exchanger (2) communicates with the material-liquid tank (1) and the water supply unit (5) through a first pipeline (6), and through a second pipeline A pipeline (7) is in communication with the steam supply device (3), and is in communication with the three-phase centrifuge (4) through a third pipeline (8);

   Preferably, the water supply unit (5) is connected to the first pipeline (6) through a distribution station and a first branch pipe (61); and the liquid-liquid tank (1) passes through the distribution station and a second branch pipe The road (62) is connected to the first pipeline (6);

   Preferably, a pressure pump (11) is further provided on the second branch pipe (62);

   Preferably, a pressure gauge (10) is further provided on the first pipeline (6);

   Preferably, the distribution station is further connected with a viewing cup (12).
3. The device according to claim 1 or 2, wherein the heat exchanger (2) is a plate heat exchanger or a tube heat exchanger.
4. The device according to claim 1 or 2, wherein a temperature monitoring device (9) is further provided on a pipeline connecting the heat exchanger (2) and the three-phase centrifuge (4).
5. A method for online instantaneous heating and separation of microbial oils and fats, characterized in that the method includes:

   (1) System preheating: Water is continuously supplied to the heat exchanger by the water supply unit, and steam is continuously supplied to the heat exchanger by the steam supply device to exchange heat between the water flow and the steam in the heat exchanger and make the water The temperature of the effluent water flow is maintained at a first predetermined temperature, and then water is introduced into the three-phase centrifuge to preheat the three-phase centrifuge, so that the three-phase centrifuge reaches a second predetermined temperature;

   (2) Material water switching: cut off the water supply of the water supply unit, and continuously supply cracking liquid from the material liquid tank to the heat exchanger, and the steam supply device continues to supply steam to the heat exchanger, and the cracking liquid stream and the steam are made in the heat exchanger. Performing heat exchange and bringing the temperature of the lysate effluent stream to a third predetermined temperature; then introducing the lysate into a preheated three-phase centrifuge for separation of the oil phase, the water phase and the solid phase to obtain microbial oils and fats;

   The lysate is a lysate of a fermentation broth of an oil-producing microorganism.
6. The method according to claim 5, wherein the first predetermined temperature, the second predetermined temperature, and the third predetermined temperature are each independently 80-95 ° C.
7. The method according to claim 5 or 6, wherein in step (1), the water supply unit continuously sends water to the heat exchanger through the first branch pipe 61 and the first pipe 6 by sending water to the distribution station. Water supply; the water supply pressure is 0.1-0.2MPa;

   In step (2), the lysate from the liquid-liquid tank enters the replacement tap water from the distribution station, and continuously supplies the lysate to the heat exchanger through the second branch pipe 62; the supply of the lysate to the heat exchanger The hydraulic pressure of the liquid is 0.1-0.2MPa;

   Preferably, after the feed water is switched, the method further includes discharging water in the distribution station, the first branch pipe 61 and the first pipe 6.
8. The method according to claim 5, wherein the lysate is an enzymatic hydrolysate of a fermentation broth of an oil-producing microorganism, and a method for preparing the enzymatic hydrolysate comprises: in a sterile environment, fermenting the broth of an oil-producing microorganism Contacting with cell wall lyase to perform enzymolysis on the oil-producing microorganisms in the fermentation broth to obtain the enzymatic hydrolysate;

   Preferably, the cell wall lyase includes alkaline protease and optionally other enzymes, and the other enzymes are among cellulase, hemicellulase, pectinase, snail enzyme, chitinase, and ligninase. At least one.
9. The method according to claim 8, wherein the conditions for the enzymolysis include: a pH of 8-10, a temperature of 40-60 ° C, a pressure of 0.02-0.05MPa, a ventilation volume of 0.2-0.6VVM, and a time of 4-15 hours;

   Preferably, the enzymolysis is performed under stirring conditions, and the stirring speed is 8-30 rpm.
10. The microbial fat prepared by the method according to any one of claims 5 to 9, wherein the microbial fat is hair oil, and the DHA content of the hair oil is greater than 35% by weight or the ARA content is greater than 35% by weight, 3 chloropropanol <350 μg / kg, anisidine value is less than 25, and peroxide value is less than 20 meq / kg.

Images