CN107801938B - Biological softening method of betel nuts - Google Patents
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- CN107801938B CN107801938B CN201711048273.3A CN201711048273A CN107801938B CN 107801938 B CN107801938 B CN 107801938B CN 201711048273 A CN201711048273 A CN 201711048273A CN 107801938 B CN107801938 B CN 107801938B
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- 230000004151 fermentation Effects 0.000 claims abstract description 52
- 244000005700 microbiome Species 0.000 claims abstract description 46
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- 238000004140 cleaning Methods 0.000 claims abstract description 15
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- 238000012545 processing Methods 0.000 claims abstract description 11
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- 239000001963 growth medium Substances 0.000 claims description 27
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 14
- 229930006000 Sucrose Natural products 0.000 claims description 14
- 229940041514 candida albicans extract Drugs 0.000 claims description 14
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- 241000894006 Bacteria Species 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 239000003480 eluent Substances 0.000 claims description 11
- 239000001888 Peptone Substances 0.000 claims description 10
- 108010080698 Peptones Proteins 0.000 claims description 10
- 238000009630 liquid culture Methods 0.000 claims description 10
- 235000019319 peptone Nutrition 0.000 claims description 10
- 230000008569 process Effects 0.000 claims description 10
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- 238000011081 inoculation Methods 0.000 claims description 3
- 239000000725 suspension Substances 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 2
- 230000002779 inactivation Effects 0.000 claims 1
- 244000080767 Areca catechu Species 0.000 abstract description 44
- 235000006226 Areca catechu Nutrition 0.000 abstract description 21
- 230000000694 effects Effects 0.000 abstract description 9
- 150000008442 polyphenolic compounds Chemical class 0.000 abstract description 6
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- 229940088598 enzyme Drugs 0.000 description 11
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- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
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- 239000002893 slag Substances 0.000 description 1
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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 OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
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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 OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
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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
- 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
- C12N1/14—Fungi; Culture media therefor
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Abstract
The invention discloses a biological softening method of areca nuts, which comprises the steps of putting the areca nuts subjected to seed development and cleaning in deep processing of the areca nuts into fermentation liquor containing microorganisms for stirring and fermentation, and softening the areca nuts by using complex enzyme generated by the microorganisms to obtain softened areca nuts; the microorganism is Eurotium cristatum strain, the mass ratio of the areca after seed development and cleaning to the fermentation liquor containing the microorganism is 1.0-25: 100, the stirring speed is 10-80 rpm, the fermentation temperature is 25-40 ℃, and the fermentation time is 12-48 h. The biological softening method has the advantages of good softening effect, high safety, capability of improving the physiological activity of the betel nut polyphenol, high edible quality and the like.
Description
Technical Field
The invention belongs to the field of agricultural product processing, relates to an areca softening method, and particularly relates to a biological softening method of areca.
Background
Betel nuts are known as the first of south medicine, have thousands of years of cultivation history in China, and are also used as special products to be eaten by local residents. The edible dry areca nuts originate from Hunan province and have a history of nearly 300 years, the annual output value of the edible dry areca nut processing industry reaches 200 hundred million yuan in 2016, and the edible dry areca nut processing industry rapidly develops at a speed increase of 15-20% every year. The edible dry areca is low in water content and high in hardness, so that fibers of the dry areca can damage oral mucosa and teeth to a certain extent in the chewing process, and the oral mucosa can be possibly fiberized to cause oral cancer. In order to improve the quality of edible dry betel nuts and improve the safety of chewing the betel nuts, the betel nut softening technology is highly concerned by the industry. In the prior art, the defects of high production cost, unsatisfactory softening effect, serious slag melting of terminal products and the like exist when different enzymes or complex enzyme preparations are adopted to soften the betel nuts. Therefore, it is urgent to find a new betel nut softening technology.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a biological softening method of areca nuts, which has good softening effect, high safety, high edible quality and capability of improving the physiological activity of areca-nut polyphenol.
In order to solve the technical problems, the invention adopts the following technical scheme.
A biological softening method of Arecae semen comprises placing Arecae semen after deep processing and washing in fermentation liquid containing microorganism, stirring, fermenting, and softening Arecae semen with complex enzyme generated by microorganism to obtain softened Arecae semen;
the microorganism is eurotium cristatum strain;
the mass ratio of the areca nuts subjected to seed development and cleaning to the fermentation liquor containing microorganisms is 1.0-25: 100, the stirring speed is 10-80 rpm, the fermentation temperature is 25-40 ℃, and the fermentation time is 12-48 h.
In the above biological softening method for areca catechu, preferably, the fermentation liquid containing microorganisms is prepared by the following processes: inoculating 1-2 rings from the inclined plane of the activated eurotium cristatum strain into 50-100 mL of liquid culture medium, performing shake culture at the rotating speed of 50-200 r/min and the culture temperature of 20-30 ℃ for 24-72 h to obtain a primary culture solution, and then performing primary culture for a certain time according to the formula
The inoculation amount of the microorganism is gradually enlarged and cultured to obtain fermentation liquor containing the microorganism.
In the above method for biologically softening areca catechu, it is preferable that the final concentration of microorganisms in the microorganism-containing fermentation broth is 106cfu/mL～1010cfu/mL。
In the above biological softening method for betel nut, preferably, the liquid medium mainly comprises sucrose, wort, yeast extract and water, and the concentration of the raw materials in water is as follows: 20 g/L-100 g/L of cane sugar, 5 g/L-25 g/L of wort and 1.0 g/L-5.0 g/L of yeast extract; the pH of the liquid culture medium is natural, and the liquid culture medium is sterilized for 15-25 min at the temperature of 115-121 ℃.
In the above biological softening method for areca catechu, preferably, the fermentation liquid containing microorganisms is prepared by the following processes: per 1cm2Adding 0.5-1 mL of peptone solution with the mass fraction of 0.1-0.5% into the test tube inclined plane to elute and collect the strains on the test tube inclined plane of the activated Eurotium cristatum strains to obtain test tube strain eluent, and then selecting 0.001mL of peptone solution as the basis of the mass fraction of the strainscm2～0.003mL/cm2Adding the test tube strain eluent on the surface of a solid culture medium, culturing at 20-30 ℃ for 24-72 h to complete primary culture, and performing step-by-step enlarged culture according to the amount of the required fermentation liquor containing microorganisms, namely, according to the amount of every 1cm2Adding 0.5-1 mL of peptone solution with the mass fraction of 0.1-0.5% to elute and collect the bacteria on the surface of the solid culture medium cultured last time to obtain the bacteria eluent on the surface of the solid culture medium, and performing cell separation according to the proportion
Adding the collected solid medium surface bacterium eluent on the surface of a newly prepared solid medium, culturing at the temperature of 20-30 ℃ for 24-72 h, repeating the amplification culture process, and finally using peptone solution with the mass fraction of 0.1-0.5% according to the proportion of 0.1mL/cm2～0.5mL/cm2Eluting the bacteria on the surface of the obtained solid culture medium, collecting the liquid, and making into final concentration of 105cfu/mL～108cfu/mL of bacterial suspension to obtain fermentation liquor containing microorganisms.
In the above biological softening method for betel nut, preferably, the solid medium mainly comprises sucrose, wort, yeast extract, agar and water, and the concentration of the raw materials in water is as follows: 20g/L to 100g/L of cane sugar, 5g/L to 25g/L of wort, 1.0g/L to 5.0g/L of yeast extract and 16g/L to 22g/L of agar; the pH of the solid culture medium is natural, and the solid culture medium is sterilized for 15-25 min at the temperature of 115-121 ℃.
In the biological softening method of areca catechu, preferably, the eurotium cristatum strain is an eurotium cristatum strain separated from black tea in Hunan, and the preservation number is CGMCC NO. 5891.
In the biological softening method of betel nuts, preferably, after stirring and fermenting, enzyme deactivation and sterilization are carried out at 90-100 ℃ for 10-20 min, and softened betel nuts are obtained after cleaning.
In the biological softening method of betel nuts, preferably, the hardness of the softened betel nuts is reduced by 30-38% compared with the hardness of the betel nuts after seed development and cleaning.
The deep processing of betel nuts generally comprises: stewing and boiling primarily processed and dried areca in lime water, etc., drying, cutting, adding bittern, and packaging. The method of the invention is to increase the amount of the active ingredients after seed raising and washing and before drying.
Compared with the prior art, the invention has the advantages that:
the invention provides a biological softening method of edible dry areca nuts, which comprises the steps of screening microorganisms, inoculating specific microorganisms for fermentation, and utilizing a complex enzyme system generated by the specific microorganisms to moderately hydrolyze cellulose and hemicellulose in the edible dry areca nuts so as to reduce the hardness of the edible dry areca nuts, thereby realizing the softening of the areca nuts, achieving the purpose of moderately controllable softness of the areca nuts, improving the edible quality and reducing the damage to oral mucosa in the chewing process of the areca nuts.
The invention has the following specific advantages:
1. by applying the microbiological principle, specific microorganisms are selected for fermentation treatment, and purification treatment on enzyme is not needed, so that the production cost is reduced.
2. The inherent characteristics of the complex enzyme generated by the microorganisms are maintained through the fermentation of the microorganisms, and the hydrolysis degree is controllable.
3. The microbial fermentation hydrolyzes cellulose and hemicellulose properly, and the edible dry areca is softer, so that the harm to the oral cavity can be reduced.
4. The eurotium cristatum strain selected by the invention has a biotransformation effect on plant polyphenol, and can improve the physiological function of betel nut polyphenol. The strain is separated from traditional fermented black tea, has important significance for the formation of the quality of the black tea, has high safety, stable enzyme system and controllable cellulose hydrolysis, can carry out biotransformation on plant polyphenol, and can improve the physiological activity of the polyphenol.
5. The hardness of the betel nut softened by the method is reduced by 30-40% compared with that of the betel nut not treated.
6. In the method of the invention, the preparation of a suitable fermentation broth has an effect on controlling the softening effect of areca catechu. The fermentation liquor is not ideal in preparation, the softening effect is not obvious, the time consumption of the softening process is prolonged, the energy consumption is increased, and the production of finished products is improved; or excessive softening, severe slagging and the like, which lead to the appearance of the finished product in the later processing process to be easily seriously deformed.
Detailed Description
The invention is further described below with reference to specific preferred embodiments, without thereby limiting the scope of protection of the invention.
The materials and equipment used in the following examples are commercially available. The Eurotium cristatum strains used in the following examples are all Eurotium cristatum (Eurotium cristatum) XU08 isolated from black tea in Hunan, and are deposited in the China general microbiological culture Collection center (CGMCC) with the collection number of CGMCC NO. 5891.
Example 1:
the invention relates to a biological softening method of areca nuts, which comprises the following steps:
(1) preparation of fermentation broth containing microorganisms
Inoculating 2 rings of activated Eurotium cristatum strain onto 50mL of liquid culture medium, culturing in a shaking incubator at 150r/min and 25 deg.C for 40h to obtain primary culture solution, gradually expanding and culturing to obtain fermentation solution containing microorganisms with final concentration of 1.0 × 108cfu/mL～9.0×108cfu/mL。
The liquid culture medium mainly comprises cane sugar, wort, yeast extract and water, and the concentration of the raw materials is as follows: sucrose 20g/L, wort 10g/L, yeast extract 2g/L, pH natural, sterilizing at 121 deg.C for 15 min.
(2) Biological softening of betel nut
Placing the betel nuts subjected to seed development and cleaning in the deep processing of the betel nuts into a fermentation tank, adding the fermentation liquor containing the microorganisms obtained in the step (1), wherein the mass ratio of the betel nuts subjected to seed development and cleaning to the fermentation liquor containing the microorganisms is 10: 100, and stirring and fermenting for 36 hours at 30rpm at the temperature of 25 ℃. After fermentation, enzyme deactivation and sterilization are carried out for 10min at 100 ℃, and softened areca nuts with moderate softness are obtained after cleaning.
The hardness of the betel nut after softening in this example was reduced by 32% as measured by texture analyzer.
Example 2:
the invention relates to a biological softening method of areca nuts, which comprises the following steps:
(1) preparation of fermentation broth containing microorganisms
Inoculating 2 rings of activated Eurotium cristatum strain onto 100mL of liquid culture medium, performing shake culture at a rotation speed of 120r/min and a culture temperature of 22 deg.C for 50h to obtain primary culture solution, gradually expanding and culturing to required fermentation solution amount according to the inoculation amount of 2% (v/v) to obtain fermentation solution containing microorganism with final concentration of 1.0 × 109cfu/mL～9.0×109cfu/mL。
The liquid culture medium mainly comprises sucrose, wort, yeast extract and water, and the concentration of the raw materials is as follows: 50g/L of sucrose, 12g/L of wort, 2g/L of yeast extract, natural pH, and sterilizing at 121 deg.C for 15 min.
(2) Biological softening of betel nut
Placing the betel nuts subjected to seed development and cleaning in the deep processing of the betel nuts into a fermentation tank, adding the fermentation liquor containing the microorganisms obtained in the step (1), wherein the mass ratio of the betel nuts subjected to seed development and cleaning to the fermentation liquor containing the microorganisms is 18: 100, and stirring and fermenting at the temperature of 30 ℃ and the rpm of 35 for 40 hours. After fermentation, inactivating enzyme and sterilizing at 100 deg.C for 10min, and cleaning to obtain softened Arecae semen.
The hardness of the betel nut after softening in this example was reduced by 35% as measured by texture analyzer.
Example 3:
the invention relates to a biological softening method of areca nuts, which comprises the following steps:
(1) preparation of fermentation broth containing microorganisms
Per 1cm2Adding 1mL of peptone solution with the mass fraction of 0.3% into the test tube inclined plane to elute and collect the strains on the test tube inclined plane of the activated Eurotium cristatum strains to obtain test tube strain eluent, and then eluting according to the mass fraction of 0.002mL/cm2Adding test tube strain eluent into the mixtureAnd (3) performing amplification culture on the surface of a solid culture medium at the culture temperature of 24 ℃ for 65h according to the amount of the fermentation liquor containing the microorganisms required by subsequent production: per 1cm2Adding 0.4mL of peptone solution with the mass fraction of 0.3% to elute and collect the bacteria on the surface of the solid culture medium cultured last time to obtain bacteria eluent, and then eluting according to the mass fraction of 0.002mL/cm2Adding the collected bacterium eluate onto the surface of newly prepared solid culture medium, culturing at 24 deg.C for 65 hr, repeating the above culture process, and adding 0.3% peptone solution at a ratio of 0.4mL/cm2Eluting the bacteria on the surface of the obtained solid culture medium, collecting the liquid, and making into final concentration of 1.0 × 106cfu/mL～9.0×106cfu/mL of the bacterial suspension to obtain the fermentation liquor containing the microorganisms.
The solid culture medium mainly comprises sucrose, wort, yeast extract, agar and water, and the concentration of the raw materials is as follows: 100g/L of sucrose, 22g/L of wort, 4g/L of yeast extract and 20g/L of agar, natural pH and sterilization at 121 ℃ for 15 min.
(2) Biological softening of betel nut
Putting the betel nuts subjected to seed development and cleaning in deep processing into a fermentation tank, adding the fermentation liquor containing the microorganisms obtained in the step (1), wherein the mass ratio of the betel nuts subjected to seed development and cleaning to the fermentation liquor containing the microorganisms is 24: 100, and stirring and fermenting for 45 hours at 70rpm at the temperature of 28 ℃. After fermentation, inactivating enzyme and sterilizing at 100 deg.C for 10min, and cleaning to obtain softened Arecae semen.
The hardness of the softened areca was reduced by 38% as measured by texture analyzer.
In addition to the above examples, it is found through a lot of experiments that the cost of the bio-softening method of the present invention is reduced by more than 15% and the softening effect is improved by more than 30% compared with the conventional enzyme softening method. Compared with the method for softening the areca nuts by adopting the high-pressure blasting method, the method for biologically softening the areca nuts has the advantages that the hardware cost investment required by the method is only about 30 percent of that of the high-pressure blasting method when the areca nuts have the same production capacity, and the softening effect can be improved.
The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. Although the present invention has been described with reference to the preferred embodiments, it is not intended to be limited thereto. Those skilled in the art can make many possible variations and modifications to the disclosed embodiments, or equivalent modifications, without departing from the spirit and scope of the invention, using the methods and techniques disclosed above. Therefore, any simple modification, equivalent replacement, equivalent change and modification made to the above embodiments according to the technical essence of the present invention are still within the scope of the protection of the technical solution of the present invention.
Claims (5)
1. A biological softening method of Arecae semen comprises placing Arecae semen after deep processing and seed development and cleaning in fermentation liquid containing microorganism, stirring and fermenting, and softening Arecae semen with complex enzyme generated by microorganism to obtain softened Arecae semen;
the microorganism is eurotium cristatum strain;
the mass ratio of the fermented and cleaned betel nuts to the fermentation liquor containing microorganisms is 1.0-25: 100, the stirring speed is 10-80 rpm, the fermentation temperature is 25-40 ℃, and the fermentation time is 12-48 hours;
the eurotium cristatum strain is an eurotium cristatum strain separated from black tea in Hunan, and the preservation number is CGMCC NO. 5891;
the fermentation liquor containing the microorganisms is prepared by the following steps: inoculating 1-2 rings from the inclined plane of the activated eurotium cristatum strain into 50-100 mL of liquid culture medium, carrying out shake culture at the rotating speed of 50-200 r/min, the culture temperature of 20-30 ℃ and the culture time of 24-72 h to obtain a primary culture solution, and then carrying out stepwise amplification culture according to the inoculation amount of 0.5-3% to obtain a fermentation solution containing microorganisms, wherein the final concentration of the microorganisms in the fermentation solution containing the microorganisms is 106cfu/mL～1010cfu/mL；
Alternatively, the microorganism-containing fermentation broth is passed throughThe following process is prepared: per 1cm2Adding 0.5-1 mL of peptone solution with the mass fraction of 0.1-0.5% into the test tube inclined plane to elute and collect the strains on the test tube inclined plane of the activated Eurotium cristatum strains to obtain test tube strain eluent, and then eluting according to the mass fraction of 0.001mL/cm2～0.003mL/cm2Adding the test tube strain eluent on the surface of a solid culture medium, culturing at 20-30 ℃ for 24-72 h to complete primary culture, and performing step-by-step enlarged culture according to the amount of the required fermentation liquor containing microorganisms, namely, according to the amount of every 1cm2Adding 0.5-1 mL of peptone solution with the mass fraction of 0.1-0.5% to elute and collect the bacteria on the surface of the solid culture medium cultured last time to obtain the bacteria eluent on the surface of the solid culture medium according to the proportion of 0.001mL/cm2〜0.003mL/cm2Adding the collected solid medium surface bacterium eluent on the surface of a newly prepared solid medium, culturing at the temperature of 20-30 ℃ for 24-72 h, repeating the amplification culture process, and finally using peptone solution with the mass fraction of 0.1-0.5% according to the proportion of 0.1mL/cm2～0.5mL/cm2Eluting the bacteria on the surface of the obtained solid culture medium, collecting the liquid, and making into final concentration of 105cfu/mL～108cfu/mL of bacterial suspension to obtain fermentation liquor containing microorganisms.
2. The method for the biological softening of betel nuts, according to claim 1, characterized in that said liquid medium consists essentially of sucrose, wort, yeast extract and water, the concentrations of the raw materials in water being as follows: 20 g/L-100 g/L of cane sugar, 5 g/L-25 g/L of wort and 1.0 g/L-5.0 g/L of yeast extract; the pH of the liquid culture medium is natural, and the liquid culture medium is sterilized for 15-25 min at the temperature of 115-121 ℃.
3. The method for the biological softening of betel nuts, according to claim 1, characterized in that said solid medium consists essentially of sucrose, wort, yeast extract, agar and water, the concentrations of the raw materials in water being as follows: 20g/L to 100g/L of cane sugar, 5g/L to 25g/L of wort, 1.0g/L to 5.0g/L of yeast extract and 16g/L to 22g/L of agar; the pH of the solid culture medium is natural, and the solid culture medium is sterilized for 15-25 min at the temperature of 115-121 ℃.
4. The method for biologically softening betel nuts according to any one of claims 1 to 3, wherein the softened betel nuts are obtained by performing enzyme inactivation and sterilization at 90 ℃ to 100 ℃ for 10min to 20min after the stirring fermentation, and then washing.
5. The method for biologically softening betel nuts according to any one of claims 1 to 3, wherein the hardness of the softened betel nuts is reduced by 30% to 38% compared to the hardness of the betel nuts after seed development and washing.
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| CN110574895A (en) * | 2019-09-09 | 2019-12-17 | 文彦然 | preparation process of flavored areca nuts |
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