CN111567777A - Probiotic brine and application thereof in processing of areca nuts - Google Patents
Probiotic brine and application thereof in processing of areca nuts Download PDFInfo
- Publication number
- CN111567777A CN111567777A CN202010367810.6A CN202010367810A CN111567777A CN 111567777 A CN111567777 A CN 111567777A CN 202010367810 A CN202010367810 A CN 202010367810A CN 111567777 A CN111567777 A CN 111567777A
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- Prior art keywords
- probiotic
- brine
- areca
- inactivated
- nuts
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Links
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- 235000018291 probiotics Nutrition 0.000 title claims abstract description 196
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- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 title claims abstract description 155
- 238000012545 processing Methods 0.000 title claims abstract description 20
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- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 39
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 30
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- 230000002378 acidificating effect Effects 0.000 description 4
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- 241000012248 Bacillus toyonensis Species 0.000 description 1
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Classifications
-
- 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
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
-
- 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
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- 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
-
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of brine preparation processes, and particularly relates to probiotic brine and application thereof in processing of areca nuts. The application of the brine in the processing of the betel nuts can improve the health care function of the brine, protect oral mucosa and the like, improve the adverse effect caused by long-time chewing of the betel nuts and ensure health.
Description
Technical Field
The invention belongs to the technical field of brine preparation processes, and particularly relates to probiotic brine and application thereof in betel nut processing.
Background
Areca catechu (Areca catechu), also called Bingmen, Renhua and Rencui, is a typical tropical economic plant and one of the four major south-China herbs. The areca contains arecoline, tannic acid and other substances, is bitter and pungent in taste, warm in nature, enters stomach and large intestine channels, and has an exciting effect when being chewed. Modern pharmacological experiments prove that the betel nut has the effects of expelling parasites, resisting viruses, treating gout, resisting fungi and the like. Betel nut enjoys the reputation of "plant chewing gum". Betel nuts are widely cultivated in china, india, srilanca, thailand, malaysia, philippines and the like.
The invention relates to a brine, which mainly refers to areca-nut brine and is brown slurry obtained by thermal reaction of calcium hydroxide and sugar (mainly maltose). Typically, sweeteners and various flavorants are also added to the slurry. When the areca is processed into an edible product, the areca needs to be subjected to processes of baking, marinating, drying, packaging and the like, wherein the areca marinade used for marinating is a key factor influencing the edible performance of the areca. At present, the commonly used areca brine is prepared by heating and decocting lime powder (generally prepared into hydrated quicklime slurry) and maltose according to the proportion, and then adding an edible sweetener and an essence perfume and mixing uniformly, but the alkalinity of the hydrated lime in the brine is easy to stimulate oral mucosa to cause oral diseases; meanwhile, the current betel nut brine does not contain functional components and has no health-care effect.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention mainly aims to provide probiotic brine.
The second purpose of the invention is to provide the application of the probiotic brine in the processing of the betel nuts.
The third purpose of the invention is to provide a preparation process of probiotic areca nuts.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a probiotic bittern comprises conventional edible bittern and probiotic.
The probiotics is a general term of microorganisms beneficial to human beings, is a bacterial product containing live bacteria and/or dead bacteria, including compositions and products thereof, and can grow and reproduce in intestinal tracts of human bodies, generate various nutrient substances, improve the digestive function of the intestinal tracts, enhance the immunity, prevent diseases and the like; in particular, the inactivated probiotics are probiotics after being inactivated by high temperature, strong acid treatment, gamma-ray or ultraviolet irradiation and the like, and the inactivated probiotics have the advantages of higher safety than live bacteria, no limit of bacteria amount, easier production and storage, high quality stability, capability of being used together with antibiotics and the like. The inactivated probiotics can enhance the immune function of human body, prevent diseases and promote the health of organism by regulating immune response reaction, adsorbing harmful substances, inhibiting pathogenic microorganisms and the like. By adding a certain amount of probiotics into the brine, the brine has the health-care effect and is beneficial to improving the utilization value of the brine.
The invention mainly refers to areca-nut brine, which is calcium-containing brine, namely calcium salt or calcium ion-containing brine, and in addition, brine in common marinated product processing, such as brine (for example, olive brine and the like) used in fruit product processing, brine (bean curd brine and the like) used in bean product processing, brine (sauce-flavor duck neck brine, brine-hoof brine and the like) used in meat product processing, and brine (salted duck egg brine, fresh egg brine and the like) used in egg product processing can be used.
"probiotic" as referred to herein includes probiotic species of the lactic acid bacteria and/or probiotic species of the non-lactic acid bacteria.
Preferably, the lactic acid bacteria probiotics are one or more of Non-Spore lactic acid bacteria (Non-Spore lactic bacteria), Spore-forming lactic acid bacteria (Spore-producing lactic bacteria), lactic acid bacteria (lactobacillus lactis) and Irregular lactic acid bacteria (Irregular lactic bacteria).
Preferably, the non-lactic acid bacteria probiotic is one or more of a propionibacterium freudenreichii, Clostridium butyricum (Clostridium butyricum), Saccharomyces boulardii (Saccharomyces bouardii), Saccharomyces cerevisiae (Saccharomyces cerevisiae), Bacillus subtilis (including Bacillus natto), Bacillus licheniformis (Bacillus licheniformis), Bacillus eastern (Bacillus toyoi).
Specifically, the bacillus-free probiotics include Lactobacillus delbrueckii, Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus reuteri, Lactobacillus casei, Lactobacillus rhamnosus, and Lactobacillus plantarum.
The probiotics of the Bacillus lactis include Bacillus coagulans (Bacillus coagulans), Bacillus laevigatus (Bacillus laevigatus), Bacillus racemosus (Bacillus racemosus) and lactobacillus inulinus (Sporolactis inulinus).
The probiotic bacteria of the genus lactococcus include Streptococcus thermophilus, Streptococcus lactis, Leuconostoc mesenteroides, Pediococcus pentosaceus, etc.
The probiotics of the irregular lactobacillus comprises Bifidobacterium breve (Bifidobacterium breve), Bifidobacterium adolescentis (Bifidobacterium adolescentis), Bifidobacterium bifidum (Bifidobacterium bifidum) and Bifidobacterium longum (Bifidobacterium longum).
Preferably, the weight ratio of the probiotic to the conventional edible brine is 1-5: 100.
Preferably, the probiotic is a live probiotic and/or a killed probiotic. The inactivated probiotics contains abundant probiotic substances, such as peptidoglycan, extracellular polysaccharide, gamma-aminobutyric acid, lactic acid, vitamins and other metabolites, and has health care effects of enhancing the immune function, improving the oral health, preventing diseases and the like.
Preferably, the probiotic is in a solid or liquid state. Specifically, the solid main probiotics are mainly in the form of bacterial powder (granules and the like), and the liquid probiotics are mainly probiotic fermentation liquor (fermentation emulsion and the like).
The invention also provides application of the probiotic brine in processing of the betel nuts.
The invention also provides a preparation process of the probiotic areca, which comprises the following steps: the probiotic brine is adopted to perform point brine or/and soaking on the pretreated areca nuts to prepare the probiotic brine. The probiotic areca brine is applied to the processing of the areca, so that the damage of edible areca to oral mucosa can be prevented, and the health-care effect of the areca is improved.
The probiotics and the brine are reasonably matched, so that the alkalinity of the areca-nut brine can be alleviated, the irritation of the brine to oral mucosa is reduced, the effect of protecting the oral mucosa is achieved, the nutritive value of the areca-nut brine can be improved, the areca-nut brine has a health-care effect, is high in nutritive value and has a utilization value.
Preferably, the probiotic is a probiotic fermentation broth, more specifically, the probiotic may be an active probiotic or a inactivated probiotic, and the probiotic fermentation broth may be an active probiotic fermentation broth or an inactivated probiotic fermentation broth. The adoption of the fermentation liquor is more convenient for the blending of the areca-nut brine.
More preferably, the probiotic bacteria comprise bacillus subtilis and lactobacillus plantarum. Specifically, the weight ratio of the bacillus subtilis to the lactobacillus plantarum is 2-4: 1.
The fiber of the betel nut is hard, and the betel nut processed by the traditional betel nut brine is easy to stab the oral mucosa during chewing to cause oral diseases; due to the treatment of sugar and lime, the phenomena of 'brine return' and 'brine return white' can occur after the brine is stored for a period of time. In the scheme, the extracellular product of the bacillus subtilis contains a large amount of gamma-polyglutamic acid (gamma-PGA), has super-strong adsorption effect on calcium ions, and can absorb the calcium ions dissociated in the areca-nut brine, so that the moisture absorption and the calcium ions are prevented from generating calcium carbonate crystals, and the application of the bacillus subtilis in the areca-nut processing can avoid the conditions of high-temperature bittern return and brine return. The lactobacillus plantarum is one of lactic acid bacteria, and is characterized in that a large amount of acidic substances can be generated, meanwhile, enzyme substances such as cellulase and the like are generated in the fermentation process, so that the fiber in the areca nut can be softened, the alkalinity of brine can be alleviated, the harm of the brine to oral mucosa can be prevented, and meanwhile, the probiotic substances in the lactobacillus plantarum can also play a health-care role.
Preferably, the probiotic brine further comprises chitosan.
The extracellular product gamma-PGA of Bacillus subtilis is an anionic polymer due to the existence of side chain carboxyl, chitosan side chain contains amino groups, and the amino groups are protonated under acidic conditions to form an electropositive polymer. After probiotics containing bacillus subtilis and lactobacillus plantarum is added into the betel nut brine, the solution is acidic due to the existence of a large amount of acidic substances in the fermentation liquor, the added chitosan has positive electricity, and the chitosan and the gamma-PGA are crosslinked due to electrostatic interaction, so that the situations of bittern return and brine return due to moisture absorption are prevented, the problem that the product appearance and the product quality are influenced due to the areca nut bittern return and the brine return is effectively solved, and the method is applied to the betel nut processing, and has good market benefit and wide prospect.
Preferably, the weight ratio of the chitosan to the probiotic brine is 1: 30-50. The chitosan with the dosage can better form hydrogel, and simultaneously, the influence of too compact gel on the mouthfeel is avoided.
Specifically, for inactivating probiotics, the soaking is to soak the pretreated areca nuts in probiotic areca nut brine at room temperature for 12-24 hours.
Further, soaking with the addition of chitosan then becomes: firstly, placing the pretreated areca in probiotic areca brine for soaking for 12-24h at room temperature, and then adding chitosan for soaking for 3-5h at room temperature. The method is characterized in that probiotic areca-nut brine is firstly used for soaking for a period of time, so that probiotic substances in the probiotic areca-nut are favorably and fully permeated into the areca-nut, and the areca-nut brine has the optimal health care value.
Specifically, for active probiotics, the soaking is to place the pretreated areca nuts in probiotic areca nut brine, adjust the pH value of the probiotic areca nut brine to 6-7, then place the probiotic areca nut brine on a shaking bed with the rotating speed of 150-;
further, in the case of adding chitosan, the following treatment is required after the fermentation: adding chitosan into the fermented probiotic areca-nut brine, stirring and uniformly mixing, and continuously fermenting for 2-3 h.
Specifically, the bittern adding step further comprises a drying step (such as freeze drying). After drying treatment, the hydrogel layer on the surface of the betel nut can keep a certain physical stimulation resistance effect, so that the protection effect on the betel nut is enhanced.
The pretreatment of the betel nuts comprises seed selection, cleaning, drying, slicing and hole pressing; the aperture of the hole pressing treatment is 0.4-0.6mm, and the hole pressing density is 2-3/cm2. The pretreatment is the basis of the processing of the betel nuts, and can improve the effect of processing the betel nuts by the probiotic fermentation broth, so that the betel nuts have the optimal nutritional value and taste.
The invention also provides a preparation process of the inactivated probiotic areca, and the specific method comprises the following steps:
s1, selecting seeds: selecting full, uniform and intact semen Arecae;
s2, cleaning: cleaning impurities and dust on the surfaces of the areca nuts;
s3, drying: spreading the cleaned Arecae semen in a drying box, and drying with hot air at 40-55 deg.C and 1-2m/s for 60-80 min;
s4, slicing: slicing the dried areca seeds by a slicing machine;
s5, adopting a hole pressing machine to press holes on the surface of the betel nut, wherein the hole diameter is 0.4-0.6mm, and the hole pressing density is 2-3/cm2；
S6, preparing inactivated probiotic fermentation liquor: culturing according to conventional culture method of various strains to obtain fermentation liquor of the strains, and then inactivating the strains to obtain the strain;
s7, preparing inactivated probiotic areca-nut brine: adding the inactivated probiotic fermentation liquor into the areca brine to obtain the inactivated probiotic areca brine, wherein the weight ratio of the inactivated probiotic fermentation liquor to the areca brine is 1-5: 100;
s8, soaking treatment: soaking the pressed Arecae semen in the above inactivated probiotic Arecae semen brine at room temperature for 12-24 hr,
when chitosan is added, the soaking treatment is as follows: firstly, placing the areca nuts subjected to the hole pressing treatment in the inactivated probiotic areca nut brine for soaking for 12-24 hours at room temperature, then adding chitosan, and soaking for 3-5 hours at room temperature; the weight ratio of the chitosan to the inactivated probiotic areca-nut brine is 1: 30-50.
S9, taking out the soaked betel nuts, and freeze-drying to obtain the inactivated probiotic betel nuts.
The invention also provides a process for fermenting areca nuts by adopting active probiotics, which comprises the following steps:
s1, pretreatment of betel nuts: the specific method is the same as the preparation process of the inactivated probiotics areca nut, and the steps are S1-S5;
s2, preparing probiotic fermentation liquor: culturing according to conventional culture method of various strains to obtain strain fermentation liquor;
s3, preparing probiotic areca-nut brine: adding the probiotic fermentation liquor into the areca brine to obtain probiotic areca brine, wherein the weight ratio of the probiotic fermentation liquor to the areca brine is 1-5: 100;
s4, fermentation of betel nuts: placing the pretreated areca in the probiotic areca brine, adjusting the pH value of the probiotic areca brine to 6-7, placing the mixture on a shaking bed at the rotation speed of 150-;
and S5, taking out the betel nuts after fermentation, and freeze-drying to prepare the betel nuts fermented by the probiotics.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides probiotic brine, which is characterized in that probiotics are added into conventional brine, and the probiotics contain abundant probiotic substances such as peptidoglycan, exopolysaccharide, gamma-aminobutyric acid, lactic acid, vitamins and the like, so that the brine has the functions of pickling, flavoring and the like, and can adsorb harmful substances, inhibit pathogenic microorganisms, promote the growth of oral probiotics, improve mucosal injury, improve immunity and the like and protect the oral cavity through the function of the probiotics. The application of the brine in the processing of the betel nuts can improve the health care function of the brine, protect oral mucosa and the like, improve the adverse effect caused by long-time chewing of the betel nuts and ensure health.
Detailed Description
The following further describes the embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The test methods used in the following experimental examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.
Example 1 a inactivated probiotic areca-nut brine, comprising the following preparation steps:
s1, preparing betel nut brine: 15kg of lime powder, 30kg of maltose, 15kg of peptone, 15kg of glucose and 90kg of water;
s2, preparing inactivated bacillus subtilis fermentation liquor: inoculating the activated bacillus subtilis into a liquid culture medium, placing the liquid culture medium in a reciprocating type shaking table, carrying out shaking culture for 18h at the temperature of 34 ℃, wherein the rotating speed of the shaking table is 110r/min, heating a fermentation liquid to 85 ℃ after fermentation is finished, and keeping the temperature for 30min for inactivation treatment, thereby obtaining an inactivated bacillus subtilis fermentation liquid; the liquid culture medium contains per liter: 10g of peptone, 5g of yeast extract, 10g of NaCl and the balance of water, wherein the pH value is 7.0;
s3, preparing inactivated lactobacillus plantarum fermentation liquor: inoculating lactobacillus plantarum into an MRS liquid culture medium, culturing for 18h at the pH of 6.5 and the temperature of 35 ℃ under oxygen introduction, heating the fermentation broth to 85 ℃ after fermentation is finished, and keeping for 30min for inactivation treatment, thereby obtaining inactivated lactobacillus plantarum fermentation broth;
s4, adding the inactivated bacillus subtilis fermentation liquor obtained in the step S2 and the inactivated lactobacillus plantarum fermentation liquor obtained in the step S2 into the areca-nut brine obtained in the step S1, and uniformly mixing to obtain the inactivated probiotic areca-nut brine, wherein the weight ratio of the inactivated probiotic (the inactivated bacillus subtilis fermentation liquor: the inactivated lactobacillus plantarum fermentation liquor: 3:1) to the areca-nut brine is 3: 100.
Embodiment 2 a inactivated probiotic areca-nut brine, which specifically comprises the following preparation steps:
s1, preparing betel nut brine: 10kg of lime powder, 20kg of maltose, 10kg of peptone, 10kg of glucose and 80kg of water;
s2, preparing inactivated bacillus subtilis fermentation liquor: inoculating activated bacillus subtilis into a liquid culture medium, placing the liquid culture medium in a reciprocating type shaking table, carrying out shaking culture for 30 hours at the temperature of 30 ℃, wherein the rotating speed of the shaking table is 80r/min, heating a fermentation liquid to 85 ℃ after fermentation is finished, and keeping the temperature for 30 minutes for inactivation treatment, thereby obtaining an inactivated bacillus subtilis fermentation liquid; the liquid culture medium contains per liter: 8g of peptone, 3g of yeast extract, 9g of NaCl and the balance of water, wherein the pH value is 6.8.
S3, preparing inactivated bifidobacterium adolescentis fermentation liquor: inoculating the activated bifidobacterium adolescentis into an MRS liquid culture medium, carrying out anaerobic culture at 37 ℃ for 48h, heating the fermentation liquor to 85 ℃ after the fermentation is finished, and keeping the temperature for 30min for inactivation treatment, thereby obtaining inactivated bifidobacterium adolescentis fermentation liquor;
s4, preparing inactivated lactobacillus bulgaricus fermentation liquor: inoculating the activated lactobacillus bulgaricus into an MRS culture medium (pH6.4), culturing for 12h at 37 ℃, heating the fermentation liquor to 85 ℃ after the fermentation is finished, and keeping for 30min for inactivation treatment, thereby obtaining the inactivated lactobacillus bulgaricus fermentation liquor;
s5, adding the inactivated bacillus subtilis fermentation liquor obtained in the step S2, the inactivated bifidobacterium adolescentis fermentation liquor obtained in the step S3 and the inactivated lactobacillus bulgaricus fermentation liquor obtained in the step S4 into the betel nut brine obtained in the step S1, and uniformly mixing to obtain the inactivated probiotic betel nut brine, wherein the weight ratio of the inactivated probiotic (inactivated bacillus subtilis fermentation liquor: inactivated bifidobacterium adolescentis fermentation liquor: inactivated lactobacillus bulgaricus fermentation liquor ═ 4:1:1) to the betel nut brine is 4: 100.
Example 3 a inactivated probiotic areca-nut brine, comprising the following preparation steps:
s1, preparing betel nut brine: 20kg of lime powder, 40kg of maltose, 20kg of peptone, 20kg of glucose and 100kg of water;
s2, preparing inactivated bacillus subtilis fermentation liquor: inoculating the activated bacillus subtilis into a liquid culture medium, placing the liquid culture medium in a reciprocating type shaking table, carrying out shaking culture at 37 ℃ for 12 hours, wherein the rotating speed of the shaking table is 150r/min, heating a fermentation liquid to 85 ℃ after fermentation is finished, and keeping the temperature for 30min for inactivation treatment, thereby obtaining an inactivated bacillus subtilis fermentation liquid; the liquid culture medium contains per liter: peptone 12g, yeast extract 8g, NaCl 11g, balance water, pH7.4.
S3, preparing inactivated lactobacillus plantarum fermentation liquor: inoculating lactobacillus plantarum into an MRS liquid culture medium, culturing for 18h at the pH of 6.5 and the temperature of 35 ℃ under oxygen introduction, heating the fermentation broth to 85 ℃ after fermentation is finished, and keeping for 30min for inactivation treatment, thereby obtaining inactivated lactobacillus plantarum fermentation broth;
s4, preparing inactivated saccharomyces cerevisiae fermentation liquor: inoculating Saccharomyces cerevisiae into seed culture medium (glucose 10g/L, peptone 5g/L, yeast extract 15g/L, sodium chloride 4g/L, pH7.0), shake-culturing at 30 deg.C and 150r/min for 24 hr to obtain seed liquid, inoculating the seed liquid into fermentation culture medium (glucose 20g/L, ammonium sulfate 8gL, KH) at an inoculation amount of 1%2PO42.5g/L，MgSO4·7H2O0.5 g/L, pH5.5), performing shaking culture at 30 ℃ for 30h at the rotating speed of 150r/min, heating the fermentation broth to 85 ℃ after the fermentation is finished, and keeping the temperature for 30min for inactivation treatment, thereby obtaining inactivated saccharomyces cerevisiae fermentation broth;
s5, preparing an inactivated streptococcus thermophilus fermentation liquid: inoculating activated Streptococcus thermophilus into lactobacillus culture medium (yeast extract 7.5g, peptone 7.5g, glucose 10g, KH)2PO42g, 100mL of tomato juice, 800.5mL of tween-tween, 900mL of distilled water and pH6.8), culturing for 48h at 42 ℃, heating the fermentation broth to 85 ℃ after the fermentation is finished, and keeping the temperature for 30min for inactivation treatment, thereby obtaining the inactivated streptococcus thermophilus fermentation broth;
s6, preparing inactivated bifidobacterium adolescentis fermentation liquor: inoculating the activated bifidobacterium adolescentis into an MRS liquid culture medium, carrying out anaerobic culture at 37 ℃ for 48h, heating the fermentation liquor to 85 ℃ after the fermentation is finished, and keeping the temperature for 30min for inactivation treatment, thereby obtaining inactivated bifidobacterium adolescentis fermentation liquor;
s7, adding the inactivated bacillus subtilis fermentation liquor obtained in the step S2, the inactivated lactobacillus plantarum fermentation liquor obtained in the step S3, the inactivated saccharomyces cerevisiae fermentation liquor obtained in the step S4, the inactivated streptococcus thermophilus fermentation liquor obtained in the step S5 and the inactivated bifidobacterium adolescentis fermentation liquor obtained in the step S6 into the betel nut brine obtained in the step S1, and uniformly mixing to obtain the inactivated probiotic betel nut brine, wherein the weight ratio of the inactivated probiotic (the inactivated bacillus subtilis fermentation liquor: the inactivated lactobacillus plantarum fermentation liquor: the inactivated saccharomyces cerevisiae fermentation liquor: the inactivated streptococcus thermophilus fermentation liquor: the inactivated bifidobacterium adolescentis fermentation liquor: 8:1:1: 1) to the betel nut brine is 5: 100.
Example 4 an active probiotic areca brine, comprising the following preparation steps:
s1, preparing betel nut brine: 15kg of lime powder, 30kg of maltose, 15kg of peptone, 15kg of glucose and 90kg of water;
s2, preparing a bacillus subtilis fermentation liquid: inoculating the activated bacillus subtilis into a liquid culture medium, placing the liquid culture medium in a reciprocating type shaking table, carrying out shaking culture for 18h at the temperature of 34 ℃, wherein the rotating speed of the shaking table is 110r/min, and obtaining bacillus subtilis fermentation liquor after fermentation is finished; the liquid culture medium contains per liter: 10g of peptone, 5g of yeast extract, 10g of NaCl and the balance of water, wherein the pH value is 7.0;
s3, preparing lactobacillus plantarum fermentation liquor: inoculating lactobacillus plantarum into an MRS liquid culture medium, culturing for 18h at the pH of 6.5 and the temperature of 35 ℃ by introducing oxygen, and obtaining inactivated lactobacillus plantarum fermentation liquor after fermentation is finished;
and S4, adding the bacillus subtilis fermentation liquor obtained in the step S2 and the lactobacillus plantarum fermentation liquor obtained in the step S2 into the betel nut brine obtained in the step S1, and uniformly mixing to obtain active probiotic betel nut brine, wherein the weight ratio of the probiotics (bacillus subtilis fermentation liquor: lactobacillus plantarum fermentation liquor-3: 1) to the betel nut brine is 2: 100.
Example 5 an active probiotic areca brine, comprising the following preparation steps:
s1, preparing betel nut brine: 10kg of lime powder, 20kg of maltose, 10kg of peptone, 10kg of glucose and 80kg of water;
s2, preparing a bacillus subtilis fermentation liquid: inoculating the activated bacillus subtilis into a liquid culture medium, placing the liquid culture medium in a reciprocating type shaking table, carrying out shaking culture for 30 hours at the temperature of 30 ℃, wherein the rotating speed of the shaking table is 80r/min, and obtaining bacillus subtilis fermentation liquor after fermentation is finished; the liquid culture medium contains per liter: 8g of peptone, 3g of yeast extract, 9g of NaCl and the balance of water, wherein the pH value is 6.8.
S3, preparing a bifidobacterium adolescentis fermentation broth: inoculating the activated bifidobacterium adolescentis into an MRS liquid culture medium, carrying out anaerobic culture at 37 ℃ for 48h, and obtaining bifidobacterium adolescentis fermentation liquor after fermentation is finished;
s4, preparing lactobacillus bulgaricus fermentation liquor: inoculating the activated lactobacillus bulgaricus into an MRS culture medium (pH6.4), culturing for 12h at 37 ℃, and obtaining lactobacillus bulgaricus fermentation liquor after fermentation is finished;
s5, adding the bacillus subtilis fermentation liquor obtained in the step S2, the bifidobacterium adolescentis fermentation liquor obtained in the step S3 and the lactobacillus bulgaricus fermentation liquor obtained in the step S4 into the betel nut brine obtained in the step S1, and uniformly mixing to obtain the active probiotic betel nut brine, wherein the weight ratio of probiotics (the bacillus subtilis fermentation liquor: the bifidobacterium adolescentis fermentation liquor: the lactobacillus bulgaricus fermentation liquor: 4:1:1) to the betel nut brine is 1: 100.
Example 6 an active probiotic areca brine, comprising the following preparation steps:
s1, preparing betel nut brine: 20kg of lime powder, 40kg of maltose, 20kg of peptone, 20kg of glucose and 100kg of water;
s2, preparing a bacillus subtilis fermentation liquid: inoculating the activated bacillus subtilis into a liquid culture medium, placing the liquid culture medium in a reciprocating type shaking table, carrying out shaking culture at 37 ℃ for 12 hours, wherein the rotating speed of the shaking table is 150r/min, and obtaining bacillus subtilis fermentation liquor after fermentation is finished; the liquid culture medium contains per liter: peptone 12g, yeast extract 8g, NaCl 11g, balance water, pH7.4.
S3, preparing lactobacillus plantarum fermentation liquor: inoculating lactobacillus plantarum into an MRS liquid culture medium, culturing for 18h at the pH of 6.5 and the temperature of 35 ℃ by introducing oxygen, and obtaining lactobacillus plantarum fermentation liquor after fermentation is finished;
s4, preparing a saccharomyces cerevisiae fermentation liquid: inoculating Saccharomyces cerevisiae into seed culture medium (glucose 10g/L, peptone 5g/L, yeast extract 15g/L, sodium chloride 4g/L, pH7.0), shake-culturing at 30 deg.C and 150r/min for 24 hr to obtain seed liquid, inoculating the seed liquid into fermentation culture medium (glucose 20g/L, ammonium sulfate 8gL, KH) at an inoculation amount of 1%2PO42.5g/L，MgSO4·7H2O0.5 g/L, pH5.5), performing shaking culture at 30 ℃ at a rotating speed of 150r/min for 30h, and obtaining a saccharomyces cerevisiae fermentation liquid after fermentation is finished;
s5, preparing a streptococcus thermophilus fermentation liquid: inoculating activated Streptococcus thermophilus into lactobacillus culture medium (yeast extract 7.5g, peptone 7.5g, glucose 10g, KH)2PO42g, tomato juice 100mL, tween-800.5 mL, distilled water 900mL, pH6.8), culturing at 42 ℃ for 48h to obtain streptococcus thermophilus fermentation liquor after fermentation is finished;
s6, preparing a bifidobacterium adolescentis fermentation broth: inoculating the activated bifidobacterium adolescentis into an MRS liquid culture medium, carrying out anaerobic culture at 37 ℃ for 48h, and obtaining bifidobacterium adolescentis fermentation liquor after fermentation is finished;
s7, adding the bacillus subtilis fermentation liquor obtained in the step S2, the lactobacillus plantarum fermentation liquor obtained in the step S3, the saccharomyces cerevisiae fermentation liquor obtained in the step S4, the streptococcus thermophilus fermentation liquor obtained in the step S5 and the bifidobacterium adolescentis fermentation liquor obtained in the step S6 into the betel nut brine obtained in the step S1, and uniformly mixing to prepare the active probiotic betel nut brine, wherein the weight ratio of the probiotics (the bacillus subtilis fermentation liquor: the lactobacillus plantarum fermentation liquor: the saccharomyces cerevisiae fermentation liquor: the streptococcus thermophilus fermentation liquor: the bifidobacterium adolescentis fermentation liquor: 8:1:1: 1) to the betel nut brine is 5: 100.
Example 7 preparation of inactivated probiotic areca
S1, selecting seeds: selecting full, uniform and intact semen Arecae;
s2, cleaning: cleaning impurities and dust on the surfaces of the areca nuts;
s3, drying: spreading the cleaned Arecae semen in a drying box, and drying with hot air at 47.5 deg.C and 1.5m/s for 70 min;
s4, slicing: slicing the softened areca seeds by a slicing machine;
s5, adopting a hole pressing machine to press holes on the surfaces of the areca nuts, wherein the hole diameter is 0.65mm, and the hole pressing density is 2.5 holes/cm2；
S6, soaking treatment: placing the pressed areca nuts in the inactivated probiotic areca nut brine in the embodiment 1 for soaking at room temperature for 18 h;
s7, taking out the soaked betel nuts, and freeze-drying to obtain the inactivated probiotic betel nuts.
Example 8 preparation of inactivated probiotic areca
S1, selecting seeds: selecting full, uniform and intact semen Arecae;
s2, cleaning: cleaning impurities and dust on the surfaces of the areca nuts;
s3, drying: spreading the cleaned Arecae semen in a drying box, and drying with hot air at 40 deg.C and wind speed of 2m/s for 80 min;
s4, slicing: slicing the softened areca seeds by a slicing machine;
s5, adopting a hole pressing machine to press holes on the surface of the betel nut, wherein the hole diameter is 0.5mm, and the hole pressing density is 3/cm2；
S6, soaking treatment: placing the pressed areca nuts in the inactivated probiotic areca nut brine in the embodiment 2 for soaking for 12 hours at room temperature;
s7, taking out the soaked betel nuts, and freeze-drying to obtain the inactivated probiotic betel nuts.
Example 9 preparation of inactivated probiotic areca
S1, selecting seeds: selecting full, uniform and intact semen Arecae;
s2, cleaning: cleaning impurities and dust on the surfaces of the areca nuts;
s3, drying: spreading the cleaned Arecae semen in a drying box, and drying with hot air at 55 deg.C and 1m/s for 60 min;
s4, slicing: slicing the softened areca seeds by a slicing machine;
s5, adopting a hole pressing machine to press holes on the surfaces of the areca nuts, wherein the hole diameter is 0.8mm, and the hole pressing density is 2/cm2；
S6, soaking treatment: placing the pressed areca nuts in the inactivated probiotic areca nut brine in the embodiment 3 for soaking for 24 hours at room temperature;
s7, taking out the soaked betel nuts, and freeze-drying to obtain the inactivated probiotic betel nuts.
Example 10A preparation method of inactivated probiotic Arecae semen
S1, selecting seeds: selecting full, uniform and intact semen Arecae;
s2, cleaning: cleaning impurities and dust on the surfaces of the areca nuts;
s3, drying: spreading the cleaned Arecae semen in a drying box, and drying with hot air at 47.5 deg.C and 1.5m/s for 70 min;
s4, slicing: slicing the softened areca seeds by a slicing machine;
s5, adopting a hole pressing machine to press holes on the surfaces of the areca nuts, wherein the hole diameter is 0.65mm, and the hole pressing density is 2.5 holes/cm2；
S6, soaking treatment: firstly, placing the areca nuts after being pressed into the holes in the inactivated probiotic areca nut brine in the embodiment 1 for soaking for 18 hours at room temperature, then adding chitosan, and soaking for 5 hours at room temperature; wherein the weight ratio of the chitosan to the betel nut brine is 1: 30;
s7, taking out the soaked betel nuts, and freeze-drying to obtain the inactivated probiotic betel nuts.
Example 11A preparation method of inactivated probiotic Arecae semen
S1, selecting seeds: selecting full, uniform and intact semen Arecae;
s2, cleaning: cleaning impurities and dust on the surfaces of the areca nuts;
s3, drying: spreading the cleaned Arecae semen in a drying box, and drying with hot air at 47.5 deg.C and 1.5m/s for 70 min;
s4, slicing: slicing the softened areca seeds by a slicing machine;
s5, adopting a hole pressing machine to press holes on the surfaces of the areca nuts, wherein the hole diameter is 0.65mm, and the hole pressing density is 2.5 holes/cm2；
S6, soaking treatment: firstly, placing the areca nuts after being pressed into the holes in the inactivated probiotic areca nut brine in the embodiment 1 for soaking for 18 hours at room temperature, then adding chitosan, and soaking for 3 hours at room temperature; wherein the weight ratio of the chitosan to the betel nut brine is 1: 50;
s7, taking out the soaked betel nuts, and freeze-drying to obtain the inactivated probiotic betel nuts.
Example 12A Process for fermenting Arecae semen with active probiotic bacteria
S1, selecting seeds: selecting full, uniform and intact semen Arecae;
s2, cleaning: cleaning impurities and dust on the surfaces of the areca nuts;
s3, drying: spreading the cleaned Arecae semen in a drying box, and drying with hot air at 47.5 deg.C and 1.5m/s for 70 min;
s4, slicing: slicing the softened areca seeds by a slicing machine;
s5, adopting a hole pressing machine to press holes on the surfaces of the areca nuts, wherein the hole diameter is 0.65mm, and the hole pressing density is 2.5 holes/cm2；
S6, adding the pressed areca nuts into the probiotic areca nut brine in the embodiment 4, adjusting the pH to 6.5, then placing the areca nuts on a shaker at the rotating speed of 175r/min, and culturing for 30 hours at the constant temperature of 36 ℃;
and S7, taking out the betel nuts after fermentation, and freeze-drying to prepare the betel nuts fermented by the probiotics.
Example 13A Process for fermenting Arecae semen with active probiotic bacteria
S1, selecting seeds: selecting full, uniform and intact semen Arecae;
s2, cleaning: cleaning impurities and dust on the surfaces of the areca nuts;
s3, drying: spreading the cleaned Arecae semen in a drying box, and drying with hot air at 40 deg.C and wind speed of 2m/s for 80 min;
s4, slicing: slicing the softened areca seeds by a slicing machine;
s5, adopting a hole pressing machine to press holes on the surface of the betel nut, wherein the hole diameter is 0.5mm, and the hole pressing density is 3/cm2；
S6, adding the pressed areca nuts into the probiotic areca nut brine in the embodiment 5, adjusting the pH to 6, then placing the mixture on a shaker at the rotation speed of 150r/min, and culturing at the constant temperature of 35 ℃ for 36 hours;
and S7, taking out the betel nuts after fermentation, and freeze-drying to prepare the betel nuts fermented by the probiotics.
Comparative example 1 preparation of inactivated probiotic areca
Comparative example 1 differs from example 7 in that: in the step S6, the addition amount of the inactivated probiotics in the areca-nut brine is different, and the weight ratio of the inactivated probiotics to the areca-nut brine in the step S6 is 1: 300.
Experimental example 1 protective action of probiotic brine on oral mucosa
(1) Test drugs: the inactivated probiotic areca brine of examples 1-3 and the activated probiotic areca brine of examples 4-6, normal saline;
(2) animals for the test: wistar rats, half male and female, weighing 140-.
(3) The test method comprises the following steps: taking 90 healthy rats fasted for 12h, dividing into 9 groups at random, each group comprises 10 rats, and culturing each group in cage with 5 rats per cage, and culturing three groups at 35 mg.kg-1The dose of the composition is obtained by intravenous injection of pentobarbital sodium into ear rim, anesthetizing for 10min, wherein 0.5mL of the live probiotic areca-nut brine of examples 1-3 is orally administered to three groups, 0.5mL of the live probiotic areca-nut brine of examples 4-6 is orally administered to another three groups, and the same amount of the live probiotic areca-nut brine of examples 4-6 is not administered to the other three groupsAdding Arecae semen bittern and normal saline for inactivating probiotic or active probiotic. Dropping every 5min for 3 times, and observing whether the oral mucosa of the mouse has red swelling, ulceration and other stimulation symptoms 10min after the third dropping, and making corresponding record.
As shown in the results in Table 1, the inactivated probiotic areca-nut brine and the activated probiotic areca-nut brine prepared by the method have no obvious stimulation phenomenon on oral mucosa, and can play a role in protecting the oral mucosa.
TABLE 1 stimulation of the oral mucosa by probiotic areca-nut brine
| Group of | Irritative symptoms |
| Example 1 inactivated probiotic Areca-nut brine | No obvious irritation symptom |
| Example 2 groups of inactivated probiotic Areca-nut brine | No obvious irritation symptom |
| Example 3 groups of inactivated probiotic Areca-nut brine | No obvious irritation symptom |
| EXAMPLE 4 active probiotic Areca brine of groups | No obvious irritation symptom |
| EXAMPLE 5 group of active probiotic Areca brine | No obvious irritation symptom |
| EXAMPLE 6 group of active probiotic Areca brine | No obvious irritation symptom |
| Areca-nut brine without adding inactivated probiotics | Red swelling and pain |
| Areca-nut brine without adding active probiotics | Red swelling and pain |
| Physiological saline | No obvious irritation symptom |
Experimental example 2 Observation and test of quality of inactivated probiotic areca
Quality observation tests were carried out using the inactivated probiotic areca catechu and commercially available areca catechu in examples 7-11 and comparative example 1 as samples, the tests were carried out in a 37 ℃ constant temperature incubator, changes in the areca catechu were observed every day, and the time for the appearance of bittern return and bittern return of the areca catechu was recorded.
As shown in the results in Table 2, the time for returning bittern and white water is greatly prolonged by using the inactivated probiotics areca nuts prepared by the method of the invention, so that the quality of the areca nuts is effectively improved. Meanwhile, in the aspect of the preparation process of the probiotic areca, the effect of inhibiting areca halogenation and brine whitening can be reduced due to the factors of too little addition of inactivated probiotic and the like.
TABLE 2 Observation of probiotic Areca catechu quality
Experimental example 3 sensory quality test of inactivated probiotic Arecae semen and active probiotic Arecae semen
To verify the sensory quality of the live probiotic and live probiotic areca nuts of the present invention, the live probiotic areca nuts of examples 7-11 and the live probiotic areca nuts of examples 12-13 were used as samples, and the areca nuts were analyzed comprehensively from both the color and hardness of the areca nuts, using commercially available areca nuts as a control. Wherein, the texture of the betel nut pulp was tested by TPA (texture profileanlysis), and the measurement was repeated 3 times per sample to average. Setting texture parameters: and the probe P/50 measures the measuring range of 2500N, the height is the height of the sample plus 10mm, the percentage of compression deformation is 40%, the detection speed is 60mm/min, the trigger force is 0.5N, the cycle number is 2 times, and the interval time is 3 s.
As shown in the results in Table 3, the live probiotic areca nuts and live probiotic areca nuts prepared by the method have good color and moderate hardness, and can prevent the problem that the oral mucosa is easy to be punctured when the areca nuts are chewed, thereby protecting the oral cavity.
TABLE 3 organoleptic qualities of probiotic Areca catechu
| Group of | Color and luster | Hardness (N) |
| Example 7 inactivated probiotic Areca catechu | Brown colour | 65 |
| Example 8 inactivated probiotic Areca catechu | Brown colour | 71 |
| Example 9 inactivated probiotic Areca catechu | Brown colour | 67 |
| Example 10 inactivated probiotic Areca catechu | Brown colour | 74 |
| Example 11 inactivated probiotic Areca catechu | Brown colour | 76 |
| Active probiotic Areca catechu of example 12 | Brown colour | 63 |
| Active probiotic Areca catechu of example 13 | Brown colour | 70 |
| Commercially available betel nut | Dark brown | 133 |
The comprehensive experimental example 1, the experimental example 2 and the experimental example 3 show that the probiotic areca-nut brine prepared by the method has no obvious stimulation to the oral cavity and has the function of protecting the oral mucosa; the application of the probiotic bacteria in the processing of the betel nuts can effectively solve the problem that the appearance and the quality of the betel nuts are affected due to the fact that the betel nuts are re-halogenated and the brine is re-whitened, and the added probiotic bacteria can enable the betel nuts to have the health-care effect.
The embodiments of the present invention have been described in detail, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.
Claims (9)
1. The probiotic brine is characterized by comprising conventional edible brine and probiotics.
2. The probiotic brine according to claim 1, wherein the probiotic comprises a probiotic of the lactic acid bacteria species and/or a probiotic of the non-lactic acid bacteria species.
3. The probiotic brine according to claim 2, wherein the lactic acid bacteria are one or more of bacillus caldus, bacillus lactis, lactococcus lactis and lactobacillus anomala.
4. The probiotic brine according to claim 2, wherein the non-lactic acid bacteria are one or more of the group consisting of propionibacterium freudenreichii, clostridium butyricum, saccharomyces boulardii, saccharomyces cerevisiae, bacillus subtilis, bacillus licheniformis and bacillus eastern ocean.
5. The probiotic brine according to claim 1, wherein the weight ratio of the probiotic to the conventional edible brine is 1-5: 100.
6. The probiotic brine according to claim 1, wherein the probiotic is live probiotic and/or inactivated probiotic.
7. A probiotic brine according to claim 1 or 6, wherein said probiotic is in solid or liquid form.
8. Use of a probiotic brine according to any one of claims 1 to 7 in the processing of areca nuts.
9. A process for preparing the probiotic areca nuts, which is characterized by being prepared by carrying out point-marinating or/and soaking on the pretreated areca nuts by using the probiotic brine as described in claim 1.
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