CN111820359A - Biodegradation method for gibberellin ketone toxin in spray-painted maize germ cypress - Google Patents
Biodegradation method for gibberellin ketone toxin in spray-painted maize germ cypress Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/21—Removal of unwanted matter, e.g. deodorisation or detoxification by heating without chemical treatment, e.g. steam treatment, cooking
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Abstract
The invention discloses a biodegradation method for gibberellin ketene toxin in guniting maize germ cypress, which adopts a mode of combining a physical method and a biodegradation method to effectively detoxify the gibberellin ketene toxin in the maize germ. According to the method, firstly, the corn germ is subjected to impurity removal operation, and then physical detoxification is carried out, so that the purity of the corn germ can be ensured, the content of the gibberellin in the corn germ is greatly reduced, the effect of a subsequent biodegradation process is greatly enhanced, the gibberellin toxin is degraded by using degrading enzyme of saccharomyces cerevisiae, 97.8% of the gibberellin toxin in the corn germ can be removed by matching with the cell wall adsorption characteristic of lactobacillus delbrueckii, and the food safety is ensured.
Description
Technical Field
The invention relates to the technical field of biological detoxification, in particular to a biodegradation method for gibberellin ketene toxin in spray-dried maize germ cypress.
Background
Zearalenone (ZEN), a mycotoxin produced mainly by fusarium fungi, is widely found in corn, barley, wheat and sorghum grains and by-products thereof, has an estrogenic effect and acts mainly on the reproductive system, thus causing hypermenorrhea in livestock, poultry and laboratory mice. Consumption of zearalenone containing foods by pregnant animals (including humans) can cause abortion, stillbirth and teratogenesis. The food made from wheat flour containing gibberellic disease can also cause poisoning symptoms of central nervous system, such as nausea, chill, headache, mental depression and ataxia.
Traditional food toxin removing methods are divided into physical methods and chemical methods, wherein the physical methods can remove part of toxins but destroy fifteen nutrients; the chemicals used in chemical processes may cause an uncertain hazard to food.
For food safety, the elimination of Zearalenone (ZEN) must ensure that the nutritional characteristics of foodstuffs, feedstuffs and human food are not altered; the zearalenone can be quickly and effectively removed; does not produce residues of toxic substances or carcinogenic/mutagenic residues; economically viable, biodegradation has been studied over the years as a means of effecting the decontamination of Zearalenone (ZEN).
Disclosure of Invention
Based on the technical problems in the background art, the invention provides a biodegradation method for the zearalenone toxin in the guniting maize germ, and solves the problems in the background art.
The invention provides the following technical scheme: a biodegradation method for gibberellin ketene toxin in spray-painted maize germ cypress, comprising the following steps:
s1, taking corn germ to be processed, carrying out water separation to remove impurities, and filtering corn germ water for later use;
s2, crushing the maize germ in the S1, and performing primary physical detoxification on the crushed maize germ under the conditions of 155-179 ℃ and steam pressure of 0.8-1.2MPa to obtain a primary maize germ detoxification product;
s3 culturing Saccharomyces cerevisiae;
(1) firstly, culturing for 48 hours in a solid culture medium, then taking a single colony, and inoculating the single colony into a liquid culture medium for shake culture;
(2) extracting a beer yeast culture product zearalenone degrading enzyme, culturing the product at 37 ℃ for 3-5 days by using beer yeast, taking a culture solution, freezing and centrifuging for 20min under the conditions of 4 ℃ and 7000-8000 r/min, or separating a supernatant and thallus cells by adopting a suction filtration method, taking the separated supernatant to obtain an extracellular crude extract, precipitating by using 30-90% ammonium sulfate, freezing and centrifuging for 10-20min under the conditions of 4 ℃ and 8000 r/min, taking a precipitate, and freezing and drying to obtain a refined zearalenone degrading enzyme;
s4 proliferation culture of Lactobacillus delbrueckii: inoculating the activated strain into proliferation culture medium at a ratio of 5-6% (V/V), and culturing at 37 deg.C for 10 hr to obtain culture solution; collecting thallus, and centrifuging the culture solution at 4 ℃ and 6000-7000r/mim for 10-15min to obtain bacterial sludge;
s5, mixing zearalenone degrading enzyme obtained in S3 and bacterial sludge in S4, mixing the mixture of the zearalenone degrading enzyme and the bacterial sludge with the primary detoxified product of the maize germ in S2, placing the mixture into a centrifugal dehydrator for centrifugal processing at the rotating speed of 4200-.
Preferably, the solid culture medium in the step (1) of the step S1 consists of 5-15% of corn flour and 70-90% of bran by weight.
Preferably, the shaking culture conditions of step (1) of step S1 are that the culture temperature is 30-40 ℃, the culture time is 24-40 h, the pH value is 5.0-9.0, and the rotation speed is 200-300 r/min.
Preferably, the composition of the liquid medium in the step (1) of step S1 is 1% Yeast Extract (Yeast Extract), 2% Peptone (Peptone), 2% Dextrose (glucose).
Preferably, the step S2 is to soften the crushed corn germ, wherein the softening temperature is controlled at 75-80 ℃, and the moisture content of the softened corn germ is controlled at 15-20%.
The invention provides a biodegradation method for gibberellin ketone toxin in guniting maize germ, which comprises the steps of firstly carrying out impurity removal operation on maize germs and then carrying out physical detoxification, ensuring the purity of the maize germs, simultaneously greatly reducing the content of the gibberellin ketone in the maize germs, greatly enhancing the effect of the subsequent biodegradation process, utilizing degrading enzyme of saccharomyces cerevisiae to carry out degradation on the gibberellin ketone toxin, and being matched with the cell wall adsorption characteristic of lactobacillus delbrueckii to remove 97.8% of the gibberellin ketone toxin in the maize germs, thereby ensuring the food safety.
Detailed Description
The technical solution of the present invention will be described in detail below with reference to specific examples.
Saccharomyces cerevisiae is also known as Saccharomyces cerevisiae, also known as Saccharomyces cerevisiae or Saccharomyces cerevisiae. Saccharomyces cerevisiae is the most widely related yeast to human, not only because it is traditionally used for making bread, steamed bread and other food and brewing wine, but also as a eukaryotic model organism in modern molecular and cellular biology, and its function is equivalent to prokaryotic model organism Escherichia coli. Saccharomyces cerevisiae is the most commonly used biological species in fermentation. The cells of Saccharomyces cerevisiae are spherical or ovoid, 5-10 μm in diameter. The propagation method is budding reproduction, the cell walls of yeasts such as saccharomyces cerevisiae and the like can better adsorb and remove ZEN, and the maximum adsorption capacity can reach 2.2 g/KG.
Example 1
A biodegradation method for gibberellin ketene toxin in spray-painted maize germ cypress comprises the following steps:
s1, taking corn germ to be processed, carrying out water separation to remove impurities, and filtering corn germ water for later use;
s2, crushing the maize germ in the S1, and subjecting the crushed maize germ to preliminary physical detoxification under the conditions of 155 ℃ and steam pressure of 0.8MPa to obtain a preliminary maize germ detoxification product;
s3 Saccharomyces cerevisiae culture:
(1) firstly, culturing for 48 hours in a solid culture medium, then taking a single colony, inoculating the single colony into a liquid culture medium, and performing shaking culture at the culture temperature of 30 ℃, the culture time of 40 hours, the pH value of 5.0 and the rotation speed of 200 r/min;
(2) extracting zearalenone degrading enzyme from a beer yeast culture product, culturing the zearalenone degrading enzyme with beer yeast at 37 ℃ for 5d, taking a culture solution, performing refrigerated centrifugation for 20min at 4 ℃ and 7000 r/min, or separating a supernatant from thallus cells by adopting a suction filtration method, taking the separated supernatant to obtain an extracellular crude extract, precipitating with 30% ammonium sulfate, performing refrigerated centrifugation for 20min at 4 ℃ and 8000 r/min, taking a precipitate, and performing refrigerated drying to obtain a refined zearalenone degrading enzyme;
s4 proliferation culture of Lactobacillus delbrueckii: inoculating the activated strain into a proliferation culture medium according to the proportion of 6% (V/V), and culturing at 37 ℃ for 10h to obtain a culture solution; collecting thallus, and centrifuging the culture solution at 4 ℃ and 7000r/mim for 15min to obtain bacterial sludge;
s5, mixing zearalenone degrading enzyme obtained in S3 and bacterial sludge in S4, mixing the mixture of the zearalenone degrading enzyme and the bacterial sludge with a preliminary corn germ detoxification product in S2, placing the mixture into a centrifugal dehydrator for centrifugal processing at the rotating speed of 5000r/min for 40min, and extracting supernatant of the mixture after centrifugation is completed to obtain detoxified corn germ.
Example 2
A biodegradation method for gibberellin ketene toxin in spray-painted maize germ cypress comprises the following steps:
s1, taking corn germ to be processed, carrying out water separation to remove impurities, and filtering corn germ water for later use;
s2, crushing the maize germ in the S1, and subjecting the crushed maize germ to preliminary physical detoxification under the conditions of 179 ℃ and 1.2MPa of steam pressure to obtain a preliminary maize germ detoxification product;
s3 Saccharomyces cerevisiae culture:
(1) firstly, culturing for 48 hours in a solid culture medium, then taking a single colony, inoculating the single colony into a liquid culture medium, and performing shaking culture at the culture temperature of 40 ℃, the culture time of 40 hours, the pH value of 9.0 and the rotating speed of 300 r/min;
(2) extracting zearalenone degrading enzyme from a beer yeast culture product, culturing the zearalenone degrading enzyme with beer yeast at 37 ℃ for 5d, taking a culture solution, performing refrigerated centrifugation for 20min at 4 ℃ and 7000 r/min, or separating a supernatant from thallus cells by adopting a suction filtration method, taking the separated supernatant to obtain an extracellular crude extract, precipitating with 90% ammonium sulfate, performing refrigerated centrifugation for 20min at 4 ℃ and 8000 r/min, taking a precipitate, and performing refrigerated drying to obtain a refined zearalenone degrading enzyme;
s4 proliferation culture of Lactobacillus delbrueckii: inoculating the activated strain into a proliferation culture medium according to the proportion of 5% (V/V), and culturing at 37 ℃ for 10h to obtain a culture solution; collecting thallus, and centrifuging the culture solution at 4 ℃ and 7000r/mim for 15min to obtain bacterial sludge;
s5, mixing zearalenone degrading enzyme obtained in S3 and bacterial sludge in S4, mixing the mixture of the zearalenone degrading enzyme and the bacterial sludge with the primary corn germ detoxification product in S2, placing the mixture into a centrifugal dehydrator for centrifugal processing at the rotating speed of 4200r/min for 40min, and extracting supernatant of the mixture after centrifugation is completed to obtain detoxified corn germ.
Example 3
A biodegradation method for gibberellin ketene toxin in spray-painted maize germ cypress comprises the following steps:
s1, taking corn germ to be processed, carrying out water separation to remove impurities, and filtering corn germ water for later use;
s2, crushing the maize germ in the S1, and subjecting the crushed maize germ to preliminary physical detoxification under the conditions of 160 ℃ and the steam pressure of 1MPa to obtain a preliminary maize germ detoxification product;
s3 Saccharomyces cerevisiae culture:
(1) firstly, culturing for 48 hours in a solid culture medium, then taking a single colony, inoculating the single colony into a liquid culture medium, and performing shaking culture at the culture temperature of 40 ℃, the culture time of 35 hours, the pH value of 70 and the rotation speed of 250 r/min;
(2) extracting zearalenone degrading enzyme from beer yeast culture product, culturing at 37 deg.C for 4d with beer yeast, collecting culture solution, freezing and centrifuging at 4 deg.C and 7500 r/min for 20min, or vacuum filtering to separate supernatant and thallus cells, collecting separated supernatant to obtain extracellular crude extractive solution, precipitating with 60% ammonium sulfate, freezing and centrifuging at 4 deg.C and 8000 r/min for 10min, collecting precipitate, and freeze drying to obtain refined zearalenone degrading enzyme;
s4 proliferation culture of Lactobacillus delbrueckii: inoculating the activated strain into proliferation culture medium at a ratio of 5-6% (V/V), and culturing at 37 deg.C for 10 hr to obtain culture solution; collecting thallus, and centrifuging the culture solution at 4 ℃ and 6500r/mim for 15min to obtain bacterial sludge;
s5, mixing zearalenone degrading enzyme obtained in S3 and bacterial sludge in S4, mixing the mixture of the zearalenone degrading enzyme and the bacterial sludge with a preliminary corn germ detoxification product in S2, placing the mixture into a centrifugal dehydrator for centrifugal processing at the rotating speed of 4500r/min for 35min, and extracting supernatant of the mixture after centrifugation is completed to obtain detoxified corn germ.
Example 4
A biodegradation method for gibberellin ketene toxin in spray-painted maize germ cypress comprises the following steps:
s1, taking corn germ to be processed, carrying out water separation to remove impurities, and filtering corn germ water for later use;
s2, crushing the maize germ in the S1, and subjecting the crushed maize germ to preliminary physical detoxification under the conditions of 170 ℃ and the steam pressure of 1.1MPa to obtain a preliminary maize germ detoxification product;
s3 Saccharomyces cerevisiae culture:
(1) firstly, culturing for 48 hours in a solid culture medium, then taking a single colony, inoculating the single colony into a liquid culture medium, and performing shaking culture at the culture temperature of 34 ℃, the culture time of 30 hours, the pH value of 7 and the rotation speed of 240 r/min;
(2) extracting zearalenone degrading enzyme from beer yeast culture product, culturing at 37 deg.C for 4d with beer yeast, collecting culture solution, freezing and centrifuging at 4 deg.C and 7400 r/min for 20min, or vacuum filtering to separate supernatant and thallus cells, collecting separated supernatant to obtain extracellular coarse extractive solution, precipitating with 60% ammonium sulfate, freezing and centrifuging at 4 deg.C and 8000 r/min for 16min, collecting precipitate, and freeze drying to obtain refined zearalenone degrading enzyme;
s4 proliferation culture of Lactobacillus delbrueckii: inoculating the activated strain into a proliferation culture medium according to the proportion of 5% (V/V), and culturing at 37 ℃ for 10h to obtain a culture solution; collecting thallus, and centrifuging the culture solution at 4 ℃ and 6000r/mim for 15min to obtain bacterial sludge;
s5, mixing zearalenone degrading enzyme obtained in S3 and bacterial sludge in S4, mixing the mixture of the zearalenone degrading enzyme and the bacterial sludge with a preliminary corn germ detoxification product in S2, placing the mixture into a centrifugal dehydrator for centrifugal processing at the rotating speed of 4700r/min for 34min, and extracting supernatant of the mixture after centrifugation is completed to obtain detoxified corn germ.
According to the method, firstly, the corn germ is subjected to impurity removal operation, and then physical detoxification is carried out, so that the purity of the corn germ can be ensured, the content of the gibberellin in the corn germ is greatly reduced, the effect of a subsequent biodegradation process is greatly enhanced, the degradation of the gibberellin by using degrading enzyme of saccharomyces cerevisiae can be carried out, 97.8% of the gibberellin in the corn germ can be removed by matching with the cell wall adsorption characteristic of lactobacillus delbrueckii, and the food safety is ensured.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (5)
1. A biodegradation method for gibberellin ketene toxin in spray-painted maize germ cypress is characterized by comprising the following steps:
s1, taking corn germ to be processed, carrying out water separation to remove impurities, and filtering corn germ water for later use;
s2, crushing the maize germ in the S1, and performing primary physical detoxification on the crushed maize germ under the conditions of 155-179 ℃ and steam pressure of 0.8-1.2MPa to obtain a primary maize germ detoxification product;
s3 Saccharomyces cerevisiae culture:
(1) firstly, culturing for 48 hours in a solid culture medium, then taking a single colony, and inoculating the single colony into a liquid culture medium for shake culture;
(2) extracting a beer yeast culture product zearalenone degrading enzyme, culturing the product at 37 ℃ for 3-5 days by using beer yeast, taking a culture solution, freezing and centrifuging for 20min under the conditions of 4 ℃ and 7000-8000 r/min, or separating a supernatant and thallus cells by adopting a suction filtration method, taking the separated supernatant to obtain an extracellular crude extract, precipitating by using 30-90% ammonium sulfate, freezing and centrifuging for 10-20min under the conditions of 4 ℃ and 8000 r/min, taking a precipitate, and freezing and drying to obtain a refined zearalenone degrading enzyme;
s4 proliferation culture of Lactobacillus delbrueckii: inoculating the activated strain into proliferation culture medium at a ratio of 5-6% (V/V), and culturing at 37 deg.C for 10 hr to obtain culture solution; collecting thallus, and centrifuging the culture solution at 4 ℃ and 6000-7000r/mim for 10-15min to obtain bacterial sludge;
s5, mixing zearalenone degrading enzyme obtained in S3 and bacterial sludge in S4, mixing the mixture of the zearalenone degrading enzyme and the bacterial sludge with the primary detoxified product of the maize germ in S2, placing the mixture into a centrifugal dehydrator for centrifugal processing at the rotating speed of 4200-.
2. The method of claim 1, wherein the method comprises the steps of: the solid culture medium in the step (1) of the step S1 comprises 5-15% of corn flour and 70-90% of bran by weight.
3. The method of claim 1, wherein the method comprises the steps of: the step (1) of the step S1 comprises the steps of culturing at the temperature of 30-40 ℃, culturing for 24-40 h, adjusting the pH value to 5.0-9.0 and rotating at the speed of 200-300 r/min.
4. The method of claim 1, wherein the method comprises the steps of: the composition of the liquid medium in the step (1) of step S1 is 1% Yeast Extract (Yeast Extract), 2% Peptone (Peptone), 2% Dextrose.
5. The method of claim 1, wherein the method comprises the steps of: and in the step S2, softening operation is carried out on the crushed maize germs, wherein the softening temperature is controlled to be 75-80 ℃, and the moisture of the softened maize germs is controlled to be 15-20%.
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| WO2009021399A1 (en) * | 2007-08-10 | 2009-02-19 | Angel Yeast Co., Ltd | Biotoxin adsorbent and its producing process |
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| WO2009021399A1 (en) * | 2007-08-10 | 2009-02-19 | Angel Yeast Co., Ltd | Biotoxin adsorbent and its producing process |
| CN103190533A (en) * | 2013-03-16 | 2013-07-10 | 赵刚绩 | Preparation method and application of zearalenone biodegradation agent |
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