A red ginseng processing method for increasing the content of rare ginsenosides and reducing pesticide residues

Abstract

The present invention provides a red ginseng processing method for increasing the content of rare ginsenosides and reducing pesticide residues, which relates to the technical field of medicinal material processing, and comprises the following steps: A, steaming fresh ginseng using atomized probiotic fermentation liquid; B, drying the steamed fresh ginseng to obtain red ginseng; the pH value of the probiotic fermentation liquid is 3.0-4.0; the OD value of the probiotic fermentation liquid is 2.0-3.0; the OD value of the probiotic fermentation liquid is 1.0-2 ...2.0-3.0; the OD value of the probiotic fermentation liquid is 2.0-3.0; the OD 600 The value is 1.2~2.5, and probiotic fermentation liquid is obtained by probiotic fermentation. The fermentation liquid with a pH of 3.0~4.0 is used as an acid catalyst instead of purified water. During the ginseng steaming process, the atomized probiotic fermentation liquid is steamed to promote the conversion of ginsenosides while degrading the organophosphorus pesticide residues in ginseng. The method is safe, efficient, green and natural, and avoids the loss of nutrients and functional components caused by boiling. The technical problems of low rare saponin content, poor efficacy and pesticide residues in the red ginseng processed and prepared in the prior art are solved.

Description

Technical Field

[0001] The invention relates to the technical field of medicinal material processing, and in particular to a red ginseng processing method for increasing the content of rare ginsenosides and reducing pesticide residues. Background Art

[0002] Red ginseng is the steamed dried root and rhizome of the cultivated product of Panax ginseng C.A.Mey. of the Araliaceae family. It is dug in autumn, washed, steamed and dried. After ginseng is processed into red ginseng, its chemical composition changes, which can enhance the efficacy; at the same time, after processing, the content of panax notoginseng in red ginseng is reduced, reducing toxicity; it has the effect of increasing efficacy and reducing toxicity. In the process of planting fresh ginseng, overdose and over-frequency use of drugs will not only lead to excessive ginseng pesticide residues, but also may cause ginseng pathogens and pests to develop drug resistance, resulting in a situation where no effective drugs are available in production. During the processing of red ginseng, the saponin components in fresh ginseng will break the glycosidic bond and ester bond to generate certain unique saponins, which are mainly reflected in the deglycosylation transformation of larger polar ginsenosides and the changes in the C-17 side chain of ginsenosides. It is a unique component of red ginseng. For example, Zhou Qile et al. systematically studied 16 ginsenosides with unique changes in the C-17 side chain in Chinese red ginseng. Zhang et al. summarized that there are 13 ginsenosides unique to red ginseng. Studies have shown that ginsenoside Rg3 and apatinib can synergistically enhance anti-cancer activity. At present, there is a relatively complete review of the research on the anti-tumor effect of the rare saponin Rg3, so this article supports the anti-tumor effect of other components of red ginseng. Rg3 also has the effects of anti-fatigue, vasodilation, improving immunity, reducing urine protein in diabetic rats, and protecting renal function. Other studies have found that ginsenoside Rk1 and ginsenoside Rg5 can inhibit the proliferation of breast cancer cells. In recent years, there are many red ginseng products on the market, as well as black ginseng products that have been steamed many times. The product quality is uneven, and the content of the main ginsenoside components varies greatly. Although Chinese patent application number 202210464992.8, a red ginseng processing method for increasing the content of rare ginsenosides, Chinese patent application number 201910196649.8, a red ginseng processing method for increasing the content of ginsenosides Rg3, Rg5 and RK1, and Chinese patent application number 202211184942.0, a red ginseng processing method for simultaneously increasing the content of ginsenosides Rb2, Rd, and Rh4, respectively provide red ginseng processing methods at different steaming temperatures, excessively high steaming temperature will destroy the nutrients in red ginseng and affect the overall effect of red ginseng.

[0003] In view of this, the present invention is proposed. Summary of the invention

[0004] The object of the present invention is to provide a red ginseng processing method for increasing the rare ginsenoside content and reducing pesticide residues, so as to solve the technical problems of low rare saponin content, poor efficacy and pesticide residues in the red ginseng processed and prepared in the prior art.

[0005] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0006] The present invention provides a red ginseng processing method for increasing the content of rare ginsenosides and reducing pesticide residues, comprising steaming fresh ginseng with atomized probiotic fermentation liquid, and then drying to obtain red ginseng;

[0007] The pH of the probiotic fermentation liquid is 3.0-4.0;

[0008] The OD of the probiotic fermentation broth 600 The value is 1.2~2.5;

[0009] The probiotics include at least one of Lactobacillus plantarum, Bifidobacterium longum or Lactobacillus gasseri.

[0010] Furthermore, steaming fresh ginseng includes circulating steaming fresh ginseng using atomized probiotic fermentation liquid;

[0011] The steaming temperature is 100°C;

[0012] The steaming time is 4 to 6 hours.

[0013] Furthermore, the preparation method of the probiotic fermentation liquid comprises inoculating the probiotics into a liquid culture medium at a ratio of 1 to 2%, anaerobic culture for 24 to 72 hours, sterilizing, and centrifuging to obtain the probiotic fermentation liquid;

[0014] The sterilization conditions include sterilization at 80-85°C for 40-50 minutes;

[0015] The centrifugal speed is 5000-15000 r / min;

[0016] The centrifugation time is 5 to 10 minutes.

[0017] Further, the liquid culture medium comprises the following components in concentrations: 5-10 g / L peptone, 5-10 g / L beef extract, 2-4 g / L yeast extract, 20-25 g / L glucose, 1-2 g / L potassium dihydrogen phosphate, 3-5 g / L sodium acetate, 1-2 g / L ammonium citrate, 0.1-0.2 g / L magnesium sulfate, 0.05-0.1 g / L manganese sulfate, and 0.05-0.1% v / v Tween-80;

[0018] The pH of the liquid culture medium is 6.2-6.4.

[0019] Furthermore, the liquid culture medium includes components with the following concentrations: 10 g / L peptone, 5 g / L beef extract, 4 g / L yeast extract, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 5 g / L sodium acetate, 2 g / L triammonium citrate, 0.2 g / L magnesium sulfate, 0.05 g / L manganese sulfate, and 0.1% v / v Tween-80.

[0020] Furthermore, the method further includes seed culture before inoculating the probiotics into the liquid culture medium at a ratio of 1-2%;

[0021] The seed culture method comprises inoculating the activated single bacterial colony into a liquid culture medium, and anaerobic culture for 18 to 24 hours to obtain a probiotic seed liquid.

[0022] Furthermore, the drying temperature is 55-75°C;

[0023] The drying time is 10 to 48 hours.

[0024] Furthermore, the fresh ginseng is at least 5 years old.

[0025] Further, when the probiotic fermentation liquid includes Lactobacillus plantarum fermentation liquid and Bifidobacterium longum fermentation liquid, the volume ratio of Lactobacillus plantarum fermentation liquid to Bifidobacterium longum fermentation liquid is 2:1;

[0026] When the probiotic fermentation liquid includes Lactobacillus plantarum fermentation liquid and Lactobacillus gasseri fermentation liquid, the volume ratio of the Lactobacillus plantarum fermentation liquid to the Lactobacillus gasseri fermentation liquid is 2:1;

[0027] When the probiotic fermentation broth includes Lactobacillus plantarum fermentation broth, Bifidobacterium longum fermentation broth and Lactobacillus gasseri fermentation broth, the volume ratio of the Lactobacillus plantarum fermentation broth, the Bifidobacterium longum fermentation broth and the Lactobacillus gasseri fermentation broth is 2:1:1.

[0028] Furthermore, the plant lactobacillus is plant lactobacillus KMHD-1 ( Lactiplantibacillus plantarum KMHD-1), deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, with the deposit number of CGMCC No.28269;

[0029] The Bifidobacterium longum is Bifidobacterium longum KMBGI 01 ( Bifidobacterium longum KMBGI 01), deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with the deposit number of CGMCC No.17175;

[0030] The Lactobacillus gasseri is Lactobacillus gasseri KMBGI 03 ( Lactobacillus gasseriKMBGI 03), deposited in the General Microbiology Center of China Microorganism Culture Collection Administration with the deposit number of CGMCC No.17177.

[0031] The present invention provides a red ginseng processing method for increasing the content of rare ginsenosides and reducing pesticide residues. The method obtains a probiotic fermentation liquid by probiotic fermentation, and uses a fermentation liquid with a pH of 3 to 4.5 as an acid catalyst instead of purified water. During the ginseng steaming process, the probiotic fermentation liquid is atomized for steaming, thereby promoting the conversion of ginsenosides and degrading the organophosphorus pesticide residues in the ginseng. The method is safe, efficient, green and natural, and avoids the loss of nutrients and functional components caused by boiling. The method solves the technical problems of low rare saponin content, poor efficacy and pesticide residues in the red ginseng processed and prepared in the prior art. DETAILED DESCRIPTION

[0032] Unless otherwise defined herein, scientific and technical terms used in conjunction with the present invention shall have the meanings commonly understood by those of ordinary skill in the art. The meaning and scope of the terms should be clear, however, in the case of any potential ambiguity, the definitions provided herein take precedence over any dictionary or external definitions. In this application, unless otherwise stated, the use of "or" means "and / or". In addition, the use of the term "including" and other forms is non-limiting.

[0033] The methods and techniques of the present invention are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated.

[0034] The present invention provides a method for processing red ginseng to increase the content of rare ginsenosides, comprising steaming fresh ginseng with atomized probiotic fermentation liquid, and then drying to obtain red ginseng;

[0035] The pH of the probiotic fermentation liquid is 3.0-4.0;

[0036] The OD of the probiotic fermentation broth 600 The value is 1.2~2.5;

[0037] The probiotics include at least one of Lactobacillus plantarum, Bifidobacterium longum or Lactobacillus gasseri.

[0038] Probiotic fermentation liquid with a pH of 3.0-4.0 is obtained by probiotic fermentation, and the fermentation liquid is used as an acid catalyst instead of purified water. During the ginseng steaming process, the atomized probiotic fermentation liquid is steamed to promote the conversion of ginsenosides while degrading the organophosphorus pesticide residues in ginseng. The method is safe, efficient, green and natural, and avoids the loss of nutrients and functional ingredients caused by boiling. The method solves the technical problems of low rare saponin content, poor efficacy and pesticide residues in the red ginseng processed and prepared in the prior art.

[0039] In some specific embodiments, steaming fresh ginseng includes circulating steaming fresh ginseng using atomized probiotic fermentation liquid;

[0040] The steaming temperature is 100°C;

[0041] The steaming time is 4 to 6 hours.

[0042] By using atomized probiotic fermentation liquid to circulate and steam fresh ginseng, the fresh ginseng is fully in contact with the probiotic fermentation liquid, the transformation of rare saponins in fresh ginseng is promoted, the efficiency of steaming fresh ginseng with probiotic fermentation liquid is improved, and organic phosphorus and other pesticide residues in fresh ginseng are removed; at the same time, the atomized acidic probiotic fermentation liquid droplets can better transform ginsenosides than water vapor, thereby improving the nutritional components and effective components of fresh ginseng to a certain extent.

[0043] In some specific embodiments, the method for preparing the probiotic fermentation broth comprises inoculating probiotics into a liquid culture medium at a ratio of 1 to 2%, anaerobic culture for 24 to 72 hours, sterilizing, and centrifuging to obtain the broth.

[0044] Among them, the sterilization conditions include sterilization at 80~85℃ for 40~50min; the centrifugal speed is 5000~15000r / min; and the centrifugal time is 5~10min.

[0045] In some specific embodiments, the liquid culture medium includes components in the following concentrations: 5~10 g / L peptone, 5~10 g / L beef extract, 2~4 g / L yeast extract, 20~25 g / L glucose, 1~2 g / L dipotassium hydrogen phosphate, 3~5 g / L sodium acetate, 1~2 g / L triammonium citrate, 0.1~0.2 g / L magnesium sulfate, 0.05~0.1 g / L manganese sulfate, and 0.05~0.1% v / v Tween-80.

[0046] Among them, the pH of the liquid culture medium is 6.2~6.4.

[0047] In some specific embodiments, the liquid culture medium includes components in the following concentrations: 10 g / L peptone, 5 g / L beef extract, 4 g / L yeast extract, 20 g / L glucose, 2 g / L dipotassium hydrogen phosphate, 5 g / L sodium acetate, 2 g / L triammonium citrate, 0.2 g / L magnesium sulfate, 0.05 g / L manganese sulfate, and 0.1% v / v Tween-80.

[0048] In some specific embodiments, the step of inoculating the probiotics into the liquid culture medium at a ratio of 1 to 2% further includes seed culture;

[0049] The seed culture method comprises inoculating the activated single bacterial colony into a liquid culture medium, and anaerobic culture for 18 to 24 hours to obtain a probiotic seed liquid.

[0050] In some specific embodiments, the drying temperature is 55-75°C;

[0051] The drying time is 10 to 48 hours.

[0052] In some specific embodiments, the fresh ginseng is at least 5 years old.

[0053] The experiment found that the effect of mixed use of Lactobacillus plantarum fermentation liquid, Bifidobacterium longum fermentation liquid and Lactobacillus gasseri fermentation liquid is better than the effect of each probiotic used alone. In some specific embodiments, when the probiotic fermentation liquid includes Lactobacillus plantarum fermentation liquid and Bifidobacterium longum fermentation liquid, the volume ratio of Lactobacillus plantarum fermentation liquid and Bifidobacterium longum fermentation liquid is 2:1; when the probiotic fermentation liquid includes Lactobacillus plantarum fermentation liquid and Lactobacillus gasseri fermentation liquid, the volume ratio of Lactobacillus plantarum fermentation liquid and Lactobacillus gasseri fermentation liquid is 2:1; when the probiotic fermentation liquid includes Lactobacillus plantarum fermentation liquid, Bifidobacterium longum fermentation liquid and Lactobacillus gasseri fermentation liquid, the volume ratio of Lactobacillus plantarum fermentation liquid, Bifidobacterium longum fermentation liquid and Lactobacillus gasseri fermentation liquid is 2:1:1.

[0054] In some specific embodiments, the Lactobacillus plantarum is Lactobacillus plantarum KMHD-1 ( Lactiplantibacillus plantarum KMHD-1), deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, with the deposit number of CGMCC No.28269;

[0055] The Bifidobacterium longum is Bifidobacterium longum KMBGI 01 ( Bifidobacterium longum KMBGI 01), deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with the deposit number of CGMCC No.17175;

[0056] The Lactobacillus gasseri is Lactobacillus gasseri KMBGI 03 ( Lactobacillus gasseri KMBGI 03), deposited in the General Microbiology Center of China Microorganism Culture Collection Administration with the deposit number of CGMCC No.17177.

[0057] Storage Instructions of Lactobacillus plantarum KMHD-1

[0058] Bacterial strain name: Lactobacillus plantarum KMHD-1;

[0059] Latin name: Lactiplantibacillus plantarum ;

[0060] Depository: General Microbiology Center, China Microbiological Culture Collection Administration;

[0061] Abbreviation of depository institution: CGMCC;

[0062] Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing;

[0063] Deposit date: August 28, 2023;

[0064] Collection Center Collection Number: CGMCC No.28269.

[0065] Bifidobacterium longum KMBGI 01 Storage Instructions

[0066] Bacterial species name: Bifidobacterium longum KMBGI 01;

[0067] Latin name: Bifidobacterium longum ;

[0068] Depository: General Microbiology Center, China Microbiological Culture Collection Administration;

[0069] Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing;

[0070] Deposit date: January 14, 2019;

[0071] Abbreviation of depository institution: CGMCC;

[0072] Collection Center Collection Number: CGMCC No.17175.

[0073] Storage Instructions of Lactobacillus gasseri KMBGI 03

[0074] Bacterial species name: Lactobacillus gasseri KMBGI 03;

[0075] Latin name: Lactobacillus gasseri ;

[0076] Depository: General Microbiology Center, China Microbiological Culture Collection Administration;

[0077] Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing;

[0078] Deposit date: January 14, 2019;

[0079] Abbreviation of depository institution: CGMCC;

[0080] Collection Center Collection Number: CGMCC No.17177.

[0081] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0082] Example 1

[0083] A novel red ginseng processing method for increasing the content of rare ginsenosides is specifically carried out according to the following steps:

[0084] 1) Prepare probiotic fermentation broth.

[0085] Lactobacillus plantarum KMHD-1 was selected as the strain.

[0086] Preparation of MRS liquid culture medium: peptone 10 g / L, beef extract 5 g / L, yeast extract powder 4 g / L, glucose 20 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 5 g / L, triammonium citrate 2 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, Tween-80 1 mL, pH 6.2, make up to 1000 mL with distilled water, and sterilize at 121°C for 20 min.

[0087] Activation of strains: After taking out the glycerol tube of probiotic strains from the refrigerator, immediately put it in a 37℃ water tank for rapid thawing, gently shake it to dissolve it within 1 minute, disinfect the surface with 75% alcohol, and put it in a sterile clean bench. Use a sterile pipette to draw about 30µL of the bacterial solution in the glycerol tube onto the plate culture medium, and use a sterile inoculation loop to inoculate the strains according to the regional division method. Place the inoculated culture medium at 37℃ for anaerobic culture for 48 hours.

[0088] Seed culture: Use a sterile inoculating loop to inoculate a single colony into a test tube containing liquid culture medium and culture anaerobically at 37°C for 24 hours.

[0089] Preparation of fermentation broth: Inoculate the liquid culture medium at a 2% inoculum volume, and culture anaerobically at 37°C for 72 hours to obtain the fermentation broth; sterilize by pasteurization at 85°C for 40 minutes. Centrifugation: Place the fermentation broth in a centrifuge, centrifuge at 10,000 r / min for 10 minutes, filter, and take the filtrate to obtain the probiotic fermentation broth, with a pH of 3.5 and an OD of 4.5. 600 is 2.3.

[0090] 2) Use atomized probiotic fermentation liquid to steam fresh ginseng.

[0091] The mixture was steamed in a 100°C environment for 4 hours using an atomizing nozzle.

[0092] 3) Steamed fresh ginseng

[0093] Five-year-old fresh ginseng is dug up, and dirt and other impurities are removed from the fresh ginseng by a ginseng cleaning machine, and the surface moisture of the fresh ginseng is drained; the cleaned fresh ginseng is placed in a red ginseng steaming cabinet, and steamed with the probiotic fermentation liquid prepared in step 1); after steaming, the steamed ginseng is dried at 55° C. for 48 hours.

[0094] Example 2

[0095] A novel red ginseng processing method for increasing the content of rare ginsenosides is specifically carried out according to the following steps:

[0096] 1) Prepare probiotic fermentation broth.

[0097] Bifidobacterium longum KMBGI 01 was selected as the bacterial strain.

[0098] Preparation of MRS liquid culture medium: peptone 10 g / L, beef extract 5 g / L, yeast extract powder 4 g / L, glucose 20 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 5 g / L, triammonium citrate 2 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, Tween-80 1 mL, pH 6.2, make up to 1000 mL with distilled water, and sterilize at 121°C for 20 min.

[0099] Activation of strains: After taking out the glycerol tube of probiotic strains from the refrigerator, immediately put it in a 37℃ water tank for rapid thawing, gently shake it to dissolve it within 1 minute, disinfect the surface with 75% alcohol, and put it in a sterile clean bench. Use a sterile pipette to draw about 30µL of the bacterial solution in the glycerol tube onto the plate culture medium, and use a sterile inoculation loop to inoculate the strains according to the regional division method. Place the inoculated culture medium at 37℃ for anaerobic culture for 48 hours.

[0100] Seed culture: Use a sterile inoculating loop to inoculate a single colony into a test tube containing liquid culture medium and culture anaerobically at 37°C for 24 hours.

[0101] Preparation of fermentation broth: Inoculate the liquid culture medium at a 2% inoculation rate, and culture anaerobically at 37°C for 96 hours to obtain the fermentation broth; sterilize by pasteurization at 85°C for 40 minutes. Centrifugation: Place the fermentation broth in a centrifuge, centrifuge at 10,000 r / min for 10 minutes, filter, and take the filtrate to obtain the probiotic fermentation broth, with a pH of 3.0 and an OD of 4.5. 600 is 1.9.

[0102] 2) Use atomized probiotic fermentation liquid to steam fresh ginseng.

[0103] The mixture was steamed in a 100°C environment for 4 hours using an atomizing nozzle.

[0104] 3) Steamed fresh ginseng

[0105] Five-year-old fresh ginseng is dug up, and dirt and other impurities are removed from the fresh ginseng by a ginseng cleaning machine, and the surface moisture of the fresh ginseng is drained; the cleaned fresh ginseng is placed in a red ginseng steaming cabinet, and steamed with the probiotic fermentation liquid prepared in step 1); after steaming, the steamed ginseng is dried at 55° C. for 48 hours.

[0106] Example 3

[0107] A novel red ginseng processing method for increasing the content of rare ginsenosides is specifically carried out according to the following steps:

[0108] 1) Prepare probiotic fermentation broth.

[0109] Lactobacillus gasseri KMBGI 03 was selected as the bacterial strain.

[0110] Preparation of MRS liquid culture medium: peptone 10 g / L, beef extract 5 g / L, yeast extract powder 4 g / L, glucose 20 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 5 g / L, triammonium citrate 2 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, Tween-80 1 mL, pH 6.2, make up to 1000 mL with distilled water, and sterilize at 121°C for 20 min.

[0111] Activation of strains: After taking out the glycerol tube of probiotic strains from the refrigerator, immediately put it in a 37℃ water tank for rapid thawing, gently shake it to dissolve it within 1 minute, disinfect the surface with 75% alcohol, and put it in a sterile clean bench. Use a sterile pipette to draw about 30µL of the bacterial solution in the glycerol tube onto the plate culture medium, and use a sterile inoculation loop to inoculate the strains according to the regional division method. Place the inoculated culture medium at 37℃ for anaerobic culture for 48 hours.

[0112] Seed culture: Use a sterile inoculating loop to inoculate a single colony into a test tube containing liquid culture medium and culture anaerobically at 37°C for 24 hours.

[0113] Preparation of fermentation broth: Inoculate the liquid culture medium at a 2% inoculum volume, and culture anaerobically at 37°C for 48 hours to obtain the fermentation broth; sterilize by pasteurization at 85°C for 40 minutes. Centrifugation: Place the fermentation broth in a centrifuge, centrifuge at 10,000 r / min for 10 minutes, filter, and take the filtrate to obtain the probiotic fermentation broth, with a pH of 4.0 and an OD of 4.0. 600 is 1.5.

[0114] 2) Use atomized probiotic fermentation liquid to steam fresh ginseng.

[0115] The mixture was steamed in a 100°C environment for 4 hours using an atomizing nozzle.

[0116] 3) Steamed fresh ginseng

[0117] Five-year-old fresh ginseng is dug up, and dirt and other impurities are removed from the fresh ginseng by a ginseng cleaning machine, and the surface moisture of the fresh ginseng is drained; the cleaned fresh ginseng is placed in a red ginseng steaming cabinet, and steamed with the probiotic fermentation liquid prepared in step 1); after steaming, the steamed ginseng is dried at 55° C. for 48 hours.

[0118] Example 4

[0119] A novel red ginseng processing method for increasing the content of rare ginsenosides is specifically carried out according to the following steps:

[0120] 1) Prepare probiotic fermentation broth.

[0121] Lactobacillus plantarum KMHD-1, Bifidobacterium longum KMBGI 01 and Lactobacillus gasseri KMBGI 03 were selected as bacterial strains.

[0122] Preparation of MRS liquid culture medium: peptone 10 g / L, beef extract 5 g / L, yeast extract powder 4 g / L, glucose 20 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 5 g / L, triammonium citrate 2 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, Tween-80 1 mL, pH 6.2, make up to 1000 mL with distilled water, and sterilize at 121°C for 20 min.

[0123] Activation of strains: After taking out the glycerol tube of probiotic strains from the refrigerator, immediately put it in a 37℃ water tank for rapid thawing, gently shake it to dissolve it within 1 minute, disinfect the surface with 75% alcohol, and put it in a sterile clean bench. Use a sterile pipette to draw about 30µL of the bacterial solution in the glycerol tube onto the plate culture medium, and use a sterile inoculation loop to inoculate the strains according to the regional division method. Place the inoculated culture medium at 37℃ for anaerobic culture for 48 hours.

[0124] Seed culture: Use a sterile inoculating loop to inoculate a single colony into a test tube containing liquid culture medium and culture anaerobically at 37°C for 24 hours.

[0125] Preparation of fermentation broth: Inoculate the liquid culture medium at a 2% inoculation rate, and culture anaerobically at 37°C for 72 hours to obtain the fermentation broth of the strain; sterilize by pasteurization at 85°C for 40 minutes. Centrifugation: Place the fermentation broth of the strain in a centrifuge, centrifuge at 10,000 r / min for 10 minutes, filter, take the filtrate, and mix the fermentation broth of Lactobacillus plantarum KMHD-1, the fermentation broth of Bifidobacterium longum KMBGI 01, and the fermentation broth of Lactobacillus gasseri KMBGI 03 at a volume ratio of 2:1:1 to obtain a mixed probiotic fermentation broth with a pH of 3.5 and an OD of 1. 600 is 2.2.

[0126] 2) Use atomized probiotic fermentation liquid to steam fresh ginseng.

[0127] The mixture was steamed in a 100°C environment for 4 hours using an atomizing nozzle.

[0128] 3) Steamed fresh ginseng

[0129] Five-year-old fresh ginseng is dug up, and dirt and other impurities are removed from the fresh ginseng by a ginseng cleaning machine, and the surface moisture of the fresh ginseng is drained; the cleaned fresh ginseng is placed in a red ginseng steaming cabinet, and steamed with the probiotic fermentation liquid prepared in step 1); after steaming, the steamed ginseng is dried at 55° C. for 48 hours.

[0130] Example 5

[0131] A novel red ginseng processing method for increasing the content of rare ginsenosides is specifically carried out according to the following steps:

[0132] 1) Prepare probiotic fermentation broth.

[0133] Lactobacillus plantarum KMHD-1 and Lactobacillus gasseri KMBGI 03 were selected as bacterial strains.

[0134] Preparation of MRS liquid culture medium: peptone 10 g / L, beef extract 5 g / L, yeast extract powder 4 g / L, glucose 20 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 5 g / L, triammonium citrate 2 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, Tween-80 1 mL, pH 6.2, make up to 1000 mL with distilled water, and sterilize at 121°C for 20 min.

[0135] Activation of strains: After taking out the glycerol tube of probiotic strains from the refrigerator, immediately put it in a 37℃ water tank for rapid thawing, gently shake it to dissolve it within 1 minute, disinfect the surface with 75% alcohol, and put it in a sterile clean bench. Use a sterile pipette to draw about 30µL of the bacterial solution in the glycerol tube onto the plate culture medium, and use a sterile inoculation loop to inoculate the strains according to the regional division method. Place the inoculated culture medium at 37℃ for anaerobic culture for 48 hours.

[0136] Seed culture: Use a sterile inoculating loop to inoculate a single colony into a test tube containing liquid culture medium and culture anaerobically at 37°C for 24 hours.

[0137] Preparation of fermentation broth: Inoculate the liquid culture medium at a 2% inoculation rate, and culture anaerobically at 37°C for 72 hours to obtain the fermentation broth of the strain; sterilize by pasteurization at 85°C for 40 minutes. Centrifugation: Place the fermentation broth of the strain in a centrifuge, centrifuge at 10,000 r / min for 10 minutes, filter, take the filtrate, and mix the fermentation broth of Lactobacillus plantarum KMHD-1 and the fermentation broth of Lactobacillus gasseri KMBGI 03 at a volume ratio of 2:1 to obtain a mixed probiotic fermentation broth with a pH of 3.5 and an OD of 1. 600 is 2.0.

[0138] 2) Use atomized probiotic fermentation liquid to steam fresh ginseng.

[0139] The mixture was steamed in a 100°C environment for 4 hours using an atomizing nozzle.

[0140] 3) Steamed fresh ginseng

[0141] Five-year-old fresh ginseng is dug up, and dirt and other impurities are removed from the fresh ginseng by a ginseng cleaning machine, and the surface moisture of the fresh ginseng is drained; the cleaned fresh ginseng is placed in a red ginseng steaming cabinet, and steamed with the probiotic fermentation liquid prepared in step 1); after steaming, the steamed ginseng is dried at 55° C. for 48 hours.

[0142] Example 6

[0143] A novel red ginseng processing method for increasing the content of rare ginsenosides is specifically carried out according to the following steps:

[0144] 1) Prepare probiotic fermentation broth.

[0145] Lactobacillus plantarum KMHD-1 and Bifidobacterium longum KMBGI 01 were selected as bacterial strains.

[0146] Preparation of MRS liquid culture medium: peptone 10 g / L, beef extract 5 g / L, yeast extract powder 4 g / L, glucose 20 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 5 g / L, triammonium citrate 2 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, Tween-80 1 mL, pH 6.2, make up to 1000 mL with distilled water, and sterilize at 121°C for 20 min.

[0147] Activation of strains: After taking out the glycerol tube of probiotic strains from the refrigerator, immediately put it in a 37℃ water tank for rapid thawing, gently shake it to dissolve it within 1 minute, disinfect the surface with 75% alcohol, and put it in a sterile clean bench. Use a sterile pipette to draw about 30µL of the bacterial solution in the glycerol tube onto the plate culture medium, and use a sterile inoculation loop to inoculate the strains according to the regional division method. Place the inoculated culture medium at 37℃ for anaerobic culture for 48 hours.

[0148] Seed culture: Use a sterile inoculating loop to inoculate a single colony into a test tube containing liquid culture medium and culture anaerobically at 37°C for 24 hours.

[0149] Preparation of fermentation broth: Inoculate the liquid culture medium at a 2% inoculation rate, and culture anaerobically at 37°C for 72 hours to obtain the fermentation broth of the strain; sterilize by pasteurization at 85°C for 40 minutes. Centrifugation: Place the fermentation broth of the strain in a centrifuge, centrifuge at 10,000 r / min for 10 minutes, filter, take the filtrate, and mix the fermentation broth of Lactobacillus plantarum KMHD-1 and the fermentation broth of Bifidobacterium longum KMBGI 01 at a volume ratio of 2:1 to obtain a mixed probiotic fermentation broth with a pH of 3.5 and an OD of 1. 600 is 1.8.

[0150] 2) Use atomized probiotic fermentation liquid to steam fresh ginseng.

[0151] The mixture was steamed in a 100°C environment for 4 hours using an atomizing nozzle.

[0152] 3) Steamed fresh ginseng

[0153] Five-year-old fresh ginseng is dug up, and dirt and other impurities are removed from the fresh ginseng by a ginseng cleaning machine, and the surface moisture of the fresh ginseng is drained; the cleaned fresh ginseng is placed in a red ginseng steaming cabinet, and steamed with the probiotic fermentation liquid prepared in step 1); after steaming, the steamed ginseng is dried at 55° C. for 48 hours.

[0154] Comparative Example 1

[0155] The difference from Example 1 is that the steam is carried out in an environment of 90° C. and by using an atomizing nozzle for atomization circulation for 4 hours.

[0156] Comparative Example 2

[0157] The difference from Example 1 is that atomized pure water is used to steam fresh ginseng.

[0158] Comparative Example 3

[0159] Different from Example 1, fresh ginseng is steamed using atomized citric acid solution, wherein the concentration of the citric acid solution is 0.25 g / L and the pH is 3.5.

[0160] Comparative Example 4

[0161] Different from Example 1, fresh ginseng is steamed using atomized acetic acid solution, wherein the concentration of the acetic acid solution is 0.01 mol / L and the pH is 3.5.

[0162] Comparative Example 5

[0163] The difference from Example 1 is that 2) fresh ginseng is steamed with atomized pure water, and before steaming the fresh ginseng, the probiotic fermentation liquid obtained in step 1) is used to soak the fresh ginseng at 25° C. for 24 hours, and then the fresh ginseng is steamed with atomized pure water.

[0164] Comparative Example 6

[0165] Different from Example 1, the pH of the probiotic fermentation liquid is 5.0.

[0166] Comparative Example 7

[0167] Different from Example 1, the pH of the probiotic fermentation liquid is 2.5.

[0168] Comparative Example 8

[0169] Different from Example 1, Lactobacillus plantarum S58 (with a deposit number of CCTCC No: M2019595) in the prior art is selected as the bacterial strain.

[0170] Comparative Example 9

[0171] The difference from Example 1 is that Lactobacillus casei CCFM1127 (with a deposit number of GDMCC No. 61497) in the prior art is selected as the bacterial strain.

[0172] Test 1 Detection of rare ginsenoside content

[0173] The saponin content of the samples was detected by ultra-high liquid chromatography analysis. The samples were selected from the red ginsengs processed in Examples 1 to 7 and Comparative Examples 1 to 8, and the saponin content of each sample was detected according to the following steps:

[0174] 1. Extraction of saponin: Take about 1g of sample powder (passed through No. 4 sieve), weigh accurately, place in Soxhlet extractor, add appropriate amount of chloroform, heat and reflux for 3 hours, discard chloroform solution, evaporate the solvent from the residue; move into a stoppered conical bottle together with the filter paper tube, accurately add 50ml of water-saturated n-butanol, stopper, leave overnight, ultrasonically treat (power 250W, frequency 50kHz) for 30 minutes, filter. Accurately measure 25ml of the filtrate, evaporate to dryness in an evaporating dish, dissolve the residue in methanol, transfer to a 5ml volumetric bottle, add methanol to the mark, shake well, filter, and take the filtrate.

[0175] 2. Ultra-high performance liquid chromatography detection: acetonitrile containing 0.1% formic acid was used as mobile phase A, and water containing 0.1% formic acid was used as mobile phase B for gradient elution; the chromatographic column was ACQUITY UPLC BEH Shield RP18 (2.1 mm×100 mm, 1.7 μm); the flow rate was 0.4 mL / min; the column temperature was 30 °C; the injection volume was 3 μL; and the autosampler temperature was 4 °C.

[0176] UPLC condition optimization: According to the spectrum, the proportion of mobile phase at different times was adjusted, and the influence of different proportions on separation and column efficiency was analyzed. After multiple adjustments and optimizations, the gradient elution program was as follows: 0-5 min, 20%-30% A; 5-12 min, 30%-33% A; 12-13 min, 33%-40% A; 13-23 min, 40%-47% A; 23-24 min, 47%-60% A; 24-26 min, 60%-65% A; 26-33 min, 65%-78% A; 33-35 min, 78%-80% A; 35-36 min, 80%-20% A; 36-41 min, 20% A. The specific elution program is shown in Table 1.

[0177] Table 1 Ultra-high performance liquid chromatography conditions

[0178]

[0179] 3. Gas chromatography detection.

[0180] 1) Sample extraction and purification.

[0181] Extraction: Weigh (5.0±0.01) g of sample into a 250 mL beaker, add 10 mL of water, let stand for 10 minutes, add 30 mL of acetonitrile and 10 mL of water after the sample is fully moistened, high-speed tissue homogenization (20,000 rpm) for 2 minutes, wash the residue with 10 mL of acetonitrile, filter, and collect the filtrate.

[0182] Purification: Transfer the filtrate to a separatory funnel, add 8g of sodium chloride, shake for 3 min, and then let stand for 10 min. The upper acetonitrile is treated with anhydrous sodium sulfate, concentrated to near dryness by rotary evaporator under reduced pressure, blown to dryness with nitrogen, and the acetone is made up to 4mL, and 2mL is transferred to a sample vial for determination of organophosphorus pesticide residues.

[0183] 2) Gas chromatograph (flame photometer) is used to determine organophosphorus pesticides.

[0184] Chromatographic column: (14%-cyanopropyl-phenyl)-methylpolysiloxane capillary column (30m×0.25mm×0.25μm), or equivalent chromatographic column;

[0185] Inlet: splitless injection, 250°C;

[0186] Carrier gas: nitrogen (>99.999%), flow rate 1.0 mL / min;

[0187] Make-up gas: hydrogen 75 mL / min, air 100 mL / min;

[0188] Column temperature: 60°C for 1 min, then increase to 200°C at 15°C / min, hold for 2 min, then increase to 250°C at 20°C / min, hold for 13 min;

[0189] Detector temperature: 250°C.

[0190] 1.0 μL of the standard mixed solution and the purified sample solution were respectively injected into the chromatograph for qualitative analysis by retention time and quantitative analysis by comparing the peak area of ​​the sample solution with that of the standard solution.

[0191] 4. Test results

[0192] 1) Calculate the contents of Rg3 (R&S), Rg5, and Rk1, as shown in Table 2.

[0193] Table 2 Contents of red ginseng saponins obtained by different methods

[0194]

[0195] Results: By comparing the rare ginsenoside content in the red ginseng processed by different embodiments and comparative examples, it was found that the rare ginsenoside content in the red ginseng processed by embodiments 1 to 6 was better than that in comparative examples 2 to 5, and embodiments 4 to 6 were better than those in embodiments 1 to 3. It can be seen that the combined use of two or more multi-species fermentation broths has a further promoting effect on the conversion of rare ginsenosides. Specifically, compared with comparative example 2, the rare ginsenoside content in embodiment 1 increased by nearly 10 times, and the use of acidic probiotic fermentation broth to steam and process red ginseng can significantly increase the rare ginsenoside content; compared with comparative examples 3 and 4, the rare ginsenoside content in embodiment 1 was improved, and the effect of steaming and processing red ginseng with acidic probiotic fermentation broth was better than that of steaming with citric acid or acetic acid solution; compared with comparative example 5, the rare ginsenoside content in embodiment 1 was significantly improved, and soaking fresh ginseng in probiotic fermentation broth could not achieve the effect of using probiotic fermentation broth to steam fresh ginseng to increase the rare ginsenoside content. Compared with Example 1, the content of rare ginsenosides in Comparative Example 6 was further increased by pH, but decreased. The content of rare ginsenosides in Comparative Example 7 was further decreased by pH, and began to show a decreasing trend. A certain pH environment can promote the increase of rare ginsenosides content, but the change of rare ginsenosides content and pH did not show a single linear change. Compared with Comparative Examples 8 and 9, the content of rare ginsenosides in Examples 1 to 5 was increased, and the effect of the probiotic fermentation broth provided by the present invention was better than that of the probiotic fermentation broth in Comparative Examples 8 and 9. Compared with Comparative Example 1, lowering the steaming temperature reduced the content of rare ginsenosides.

[0196] 2) The gas chromatograph test results are shown in Table 3.

[0197] Table 3 Gas chromatograph test results

[0198]

[0199] Results: By comparing the degradation rates of organophosphorus pesticides in red ginseng prepared from different embodiments and comparative examples, it was found that the processing methods of embodiments 1 to 6 not only increased the content of rare ginsenosides, but also increased the degradation rate of organophosphorus pesticides and reduced the pesticide residues in red ginseng.

[0200] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for processing red ginseng to increase the content of rare ginsenosides and reduce pesticide residues, characterized in that: The method comprises steaming fresh ginseng using atomized probiotic fermentation liquid, and then drying the fresh ginseng to obtain red ginseng; The pH of the probiotic fermentation liquid is 3.0-4.0; The OD of the probiotic fermentation broth 600 The value is 1.2~2.5; The probiotics include at least one of Lactobacillus plantarum, Bifidobacterium longum or Lactobacillus gasseri; The preparation method of the probiotic fermentation liquid comprises the following steps: inoculating the probiotics into a liquid culture medium at a ratio of 1 to 2%, anaerobic culturing for 24 to 72 hours, sterilizing, and centrifuging to obtain the probiotic fermentation liquid.

2. The red ginseng processing method according to claim 1, characterized in that: Steaming fresh ginseng includes utilizing atomized probiotic fermentation liquid to circulate and steam the fresh ginseng; The steaming temperature is 100°C; The steaming time is 4 to 6 hours.

3. The red ginseng processing method according to claim 1, characterized in that: The sterilization conditions include sterilization at 80-85°C for 40-50 minutes; The centrifugal speed is 5000-15000 r / min; The centrifugation time is 5 to 10 minutes.

4. The red ginseng processing method according to claim 3, characterized in that: The liquid culture medium comprises the following components in concentrations: 5-10 g / L of peptone, 5-10 g / L of beef extract, 2-4 g / L of yeast extract, 20-25 g / L of glucose, 1-2 g / L of dipotassium hydrogen phosphate, 3-5 g / L of sodium acetate, 1-2 g / L of triammonium citrate, 0.1-0.2 g / L of magnesium sulfate, 0.05-0.1 g / L of manganese sulfate, and 0.05-0.1% v / v Tween-80; The pH of the liquid culture medium is 6.2-6.

4.

5. The red ginseng processing method according to claim 4, characterized in that: The liquid culture medium includes components with the following concentrations: 10 g / L of peptone, 5 g / L of beef extract, 4 g / L of yeast extract powder, 20 g / L of glucose, 2 g / L of dipotassium hydrogen phosphate, 5 g / L of sodium acetate, 2 g / L of triammonium citrate, 0.2 g / L of magnesium sulfate, 0.05 g / L of manganese sulfate, and 0.1% v / v Tween-80.

6. The red ginseng processing method according to claim 3, characterized in that: The method further includes seed culture before inoculating the probiotics into the liquid culture medium at a ratio of 1-2%; The seed culture method comprises inoculating the activated single bacterial colony into a liquid culture medium, and anaerobic culture for 18 to 24 hours to obtain a probiotic seed liquid.

7. The red ginseng processing method according to claim 1, characterized in that: The drying temperature is 55-75°C; The drying time is 10 to 48 hours.

8. The red ginseng processing method according to claim 7, characterized in that: The fresh ginseng is fresh ginseng that is at least 5 years old.

9. The red ginseng processing method according to any one of claims 1 to 8, characterized in that: When the probiotic fermentation liquid includes Lactobacillus plantarum fermentation liquid and Bifidobacterium longum fermentation liquid, the volume ratio of the Lactobacillus plantarum fermentation liquid to the Bifidobacterium longum fermentation liquid is 2:1; When the probiotic fermentation liquid includes Lactobacillus plantarum fermentation liquid and Lactobacillus gasseri fermentation liquid, the volume ratio of the Lactobacillus plantarum fermentation liquid to the Lactobacillus gasseri fermentation liquid is 2:1; When the probiotic fermentation broth includes Lactobacillus plantarum fermentation broth, Bifidobacterium longum fermentation broth and Lactobacillus gasseri fermentation broth, the volume ratio of the Lactobacillus plantarum fermentation broth, the Bifidobacterium longum fermentation broth and the Lactobacillus gasseri fermentation broth is 2:1:

1.

10. The red ginseng processing method according to claim 9, characterized in that: The plant lactobacillus is plant lactobacillus KMHD-1 ( Lactiplantibacillus plantarum KMHD-1), deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, with the deposit number of CGMCC No.28269; The Bifidobacterium longum is Bifidobacterium longum KMBGI 01 ( Bifidobacterium longum KMBGI 01), deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with the deposit number of CGMCC No.17175; The Lactobacillus gasseri is Lactobacillus gasseri KMBGI 03 ( Lactobacillus gasseri KMBGI 03), deposited in the General Microbiology Center of China Microorganism Culture Collection Administration with the deposit number of CGMCC No.17177.

Images