CN110938552A - Method for degrading herb tea residue and preparing aspergillus niger - Google Patents
Method for degrading herb tea residue and preparing aspergillus niger Download PDFInfo
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- 241000228245 Aspergillus niger Species 0.000 title claims abstract description 43
- 241001122767 Theaceae Species 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 25
- 230000000593 degrading effect Effects 0.000 title abstract description 5
- 235000013616 tea Nutrition 0.000 claims abstract description 40
- 238000000855 fermentation Methods 0.000 claims abstract description 28
- 230000004151 fermentation Effects 0.000 claims abstract description 28
- 235000015092 herbal tea Nutrition 0.000 claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000002791 soaking Methods 0.000 claims abstract description 3
- 241000628997 Flos Species 0.000 claims description 15
- 239000000725 suspension Substances 0.000 claims description 15
- 230000001580 bacterial effect Effects 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 230000003213 activating effect Effects 0.000 claims description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 239000002609 medium Substances 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 5
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 3
- 239000002054 inoculum Substances 0.000 claims 1
- 230000015556 catabolic process Effects 0.000 abstract description 9
- 238000006731 degradation reaction Methods 0.000 abstract description 9
- 238000011081 inoculation Methods 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 3
- 238000006065 biodegradation reaction Methods 0.000 abstract description 2
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 6
- 239000000243 solution Substances 0.000 description 4
- 230000028070 sporulation Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 229940023509 aspergillus niger preparation Drugs 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000000643 oven drying Methods 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000205585 Aquilegia canadensis Species 0.000 description 1
- 241000235349 Ascomycota Species 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 235000007516 Chrysanthemum Nutrition 0.000 description 1
- 244000189548 Chrysanthemum x morifolium Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 244000303040 Glycyrrhiza glabra Species 0.000 description 1
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 description 1
- 244000235756 Microcos paniculata Species 0.000 description 1
- 241001646828 Platostoma chinense Species 0.000 description 1
- 244000215777 Plumeria rubra Species 0.000 description 1
- 235000013087 Plumeria rubra Nutrition 0.000 description 1
- 241000222383 Polyporales Species 0.000 description 1
- 244000179560 Prunella vulgaris Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 235000011477 liquorice Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 235000012015 potatoes Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 235000015067 sauces Nutrition 0.000 description 1
- 235000008113 selfheal Nutrition 0.000 description 1
- 108010027322 single cell proteins Proteins 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 235000021419 vinegar Nutrition 0.000 description 1
- 239000000052 vinegar Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 235000014101 wine Nutrition 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
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- Tropical Medicine & Parasitology (AREA)
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Abstract
The invention discloses a method for degrading herbal tea residues and preparing aspergillus niger, wherein when the aspergillus niger is produced by using the herbal tea residues, the concentration, the inoculation amount and the water content of a strain and the pH value, the fermentation duration and the fermentation temperature of a proper soaking solution are set, so that the optimal fermentation effect can be obtained, the aspergillus niger spore yield is optimal, and the degradation rate of the herbal tea residues is higher. The invention discloses a biodegradation process for fermenting cold tea leaves by using aspergillus niger for the first time, discloses a high-efficiency and low-cost method for producing aspergillus niger spores by using the cold tea leaves for the first time, and has important significance in the problem of pollution of the cold tea leaves and the aspergillus niger production process.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to a method for degrading herb tea residues and preparing aspergillus niger.
Background
The herbal tea is prepared by decocting 7 kinds of plant medicines including frangipani, honeysuckle, chrysanthemum, mesona chinensis benth, selfheal, liquorice and microcos paniculata. In order to retain the flavor components in the herbal tea as much as possible, the herbal tea is extracted at a lower temperature mostly, and the extraction times are few, so that the herbal tea residues and the original medicinal materials have similar nutrients and active components. Therefore, the herb tea residue has better nutritional and medicinal values and is a biological resource which is not fully utilized. In recent years, with the rapid development of the herbal tea industry, the discharge of the herbal tea residues is increased year by year, and the daily yield of the herbal tea residues reaches 680 tons. The traditional herb tea dregs are mostly treated by direct stacking, landfill and other modes, which causes serious resource waste and environmental pollution. Due to the limitation of the field, the herb tea residue cannot be processed in time, and even the herb tea residue has great influence on the production of enterprises. The recycling and high-value utilization of herb tea dregs become important problems which must be faced by enterprises and society.
Aspergillus niger (Aspergillus niger) belongs to the family of Moniliaceae, the subdivision Ascomycetes, Aphyllophorales, a common, safe and non-toxic fungus of the genus Aspergillus, and is widely distributed in grains, vegetable products and soil all over the world. Aspergillus niger is a main industrial strain for making sauce, wine, vinegar and saccharified feed, is a main industrial strain for producing enzyme preparations and single-cell proteins, and is also an important feed additive, but the high price of the Aspergillus niger preparation limits the wide application of the Aspergillus niger preparation in industry and agriculture.
According to the existing problems, the invention aims to develop a process method for producing aspergillus niger and spores thereof at high efficiency and low cost by microbial degradation of cold tea leaves.
Disclosure of Invention
The invention aims to provide a method for degrading herbal tea residues and producing aspergillus niger spores by aspergillus niger fermentation.
The technical scheme adopted by the invention is as follows:
in a first aspect of the present invention, there is provided a method for preparing aspergillus niger, comprising the steps of: preparing aspergillus niger strains into a bacterial suspension, and inoculating the bacterial suspension to a herbal tea residue culture medium for fermentation; the herb tea residue comprises flos Plumeriae Acutifoliae, flos Lonicerae, flos Chrysanthemi, herba mesonae chinensis, Prunellae Spica, Glycyrrhrizae radix and folium Microcorii Paniculati.
According to the embodiment of the invention, the method further comprises the steps of activating and passaging the aspergillus niger in advance, wherein the activating is to inoculate the aspergillus niger to the PDA solid medium for culture.
According to the embodiment of the invention, the fermentation condition is 50-90% of humidity and 28-37 ℃, and the culture is carried out for 24-240 h;
according to the embodiment of the invention, the passage is 1-3 times.
According to an embodiment of the invention, the bacterial suspension has a concentration of 106～109cfu/mL, preferably at a bacterial suspension concentration of 2X 107cfu/mL Aspergillus niger suspension.
According to the embodiment of the invention, the inoculation amount is 5-25%, the fermentation temperature is 28-37 ℃, and the fermentation time is 1-10 days.
According to the embodiment of the invention, the water content during fermentation is 65-85%, and the pH of the soak solution is 5-10.
According to an embodiment of the present invention, the herbal tea grounds medium comprises: 1-5 parts of cold tea residue powder, 0.04-0.20 part of ammonium sulfate, 0-0.1 part of glucose, 0.005-0.02 part of monopotassium phosphate, 0.002-0.020 part of dipotassium hydrogen phosphate and 5.5-8.5 parts of water.
According to the embodiment of the invention, 2 parts of cold tea leaves powder are added with 0.08 part of ammonium sulfate, 0.04 part of glucose, 0.01 part of monopotassium phosphate, 0.008 part of dipotassium phosphate and 8 parts of water.
According to the embodiment of the invention, the preparation method of the cool tea residue powder comprises the following steps: mixing flos Plumeriae Acutifoliae, flos Lonicerae, flos Chrysanthemi, herba mesonae chinensis, Prunellae Spica, Glycyrrhrizae radix and folium Microcorii Paniculati, decocting to obtain herbal tea residue, oven drying, and pulverizing.
The invention has the beneficial effects that:
the invention fully utilizes the residual value of the cool tea leaves and can effectively degrade a large amount of the cool tea leaves. Meanwhile, the black mold is produced by utilizing the cool tea leaves: the dosage of ammonium sulfate is 4%, the dosage of glucose is 2%, and the strain concentration is 2 × 107cfu/mL, when the inoculation amount is 10%, the water content is 80%, the pH of the soak solution is 8, the fermentation temperature is 31 ℃, and the optimal fermentation effect can be obtained after fermentation is carried out for 7 days. The yield of Aspergillus niger spores can reach (1149.14 +/-345.48) multiplied by 109cfu/g, herb tea residue degradationThe ratio was 19.58% + -0.56%. Provides a thought for the large-scale production of the Aspergillus niger. The invention discloses a biodegradation process for fermenting cold tea leaves by using aspergillus niger for the first time, discloses an efficient and cheap method for producing aspergillus niger spores by using the cold tea leaves for the first time, and has important significance for solving the problem of pollution of the cold tea leaves and efficiently and cheaply producing the aspergillus niger.
Detailed Description
The following embodiments are further illustrated by reference to the following specific examples:
preliminary preparation
1. Activating strains:
(1) activating the strain on a PDA solid culture medium, and culturing at 30 deg.C for 120h at 80% humidity in an electrothermal constant-temperature constant-humidity incubator;
(2) after two successive passages of the Aspergillus niger strain, 2X 10 is made up by using physiological saline7cfu/mL of bacterial suspension;
PDA solid medium: peeling 200g of potatoes, cutting into blocks, boiling for 30min, filtering by using gauze, adding 20g of cane sugar, and supplementing water to 1000mL after dissolving. Adding agar 20g, heating to dissolve, and wet-heat sterilizing at 121 deg.C for 20 min.
2. Preparing a herbal tea residue culture medium:
taking 6-8 parts of water, adding 0.08 part of ammonium sulfate, 0.04 part of glucose, 0.01 part of potassium dihydrogen phosphate and 0.008 part of dipotassium hydrogen phosphate, adjusting the pH to 7.0-9.0 by using potassium hydroxide, adding 2 parts of cold tea leaves, and carrying out damp-heat sterilization at 121 ℃ for 20 min;
cooling tea leaves: mixing 7 kinds of plant medicines including flos Plumeriae Acutifoliae, flos Lonicerae, flos Chrysanthemi, herba mesonae chinensis, Prunellae Spica, Glycyrrhrizae radix and folium Microcoris Paniculatae, and decocting to obtain herb tea residue.
The specific operations of examples 1 to 7 were as follows:
method for preparing aspergillus niger
Inoculating according to the inoculation amount of 10% (namely inoculating 0.1mL of Aspergillus niger spore suspension to 1g of dry herb tea residue) and inoculating aspergillus niger on the herb tea residue culture medium, wherein the specific fermentation conditions are shown in Table 1.
Measuring the number of aspergillus niger spores after fermentation and calculating the degradation of the cool tea leaves:
1. after fermentation, adding 15mL of physiological saline into the fermentation product;
2. shaking, and diluting 20 μ L of the suspension to 1.25 × 105After doubling, taking 50 mu L of coated plate, culturing, counting viable bacteria, and calculating the sporulation amount;
3. centrifuging the rest suspension at 15000g for 10 min, collecting precipitate, oven drying, and weighing;
4. and calculating sporulation quantity and degradation rate, wherein the degradation rate formula is (total weight-residual weight)/total weight.
The settings, spore yields and degradation rates of examples 1-7 are shown in Table 1:
TABLE 1 fermentation factor settings, sporulation yields, degradation rates of examples 1-7
Note: the water content is the percentage content of water when the system only contains the cold tea leaves and the water.
The soaking solution is a solution which is prepared by adding potassium dihydrogen phosphate and dipotassium hydrogen phosphate into a certain volume of water according to the proportion requirement and then adjusting the pH value to a fixed value by using 3mol/L potassium hydroxide.
Example 8
Method for preparing aspergillus niger
Inoculating according to the inoculation amount of 10% (namely inoculating 0.1mL of Aspergillus niger spore suspension to 1g of dry herb tea residue) and inoculating Aspergillus niger on the herb tea residue culture medium, wherein the water content in a fermentation system is 80%, the pH value of an immersion liquid is 8.0, the fermentation temperature is 31 ℃, and the fermentation time is 7 days.
As a result: the results of the analysis and treatment experiments showed that the method of example 8 gave a Aspergillus niger spore yield of (1149.14. + -. 345.48). times.109cfu/g; the degradation rate of the cool tea leaves is 19.58% +/-0.56%.
Example 9
Method for preparing aspergillus niger
Inoculating according to the inoculation amount of 10% (namely inoculating 0.1mL of Aspergillus niger spore suspension to 1g of dry herb tea residue) and inoculating Aspergillus niger on the herb tea residue culture medium, wherein the water content in a fermentation system is 60%, the pH value of an immersion liquid is 8.0, the fermentation temperature is 37 ℃, and the fermentation time is 5 days.
As a result: the results of the analysis treatment showed that the spore yield was (478.55. + -. 152.37). times.10 using the method of example 99cfu/g; the degradation rate of the cool tea leaves is 24.49% +/-1.29%.
According to the arrangement of the above embodiment, the embodiment 8 and the embodiment 9 are found to have the best effect, and can effectively degrade the cool tea leaves and produce aspergillus niger spores.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A method for preparing Aspergillus niger, comprising the following steps: preparing aspergillus niger into a bacterial suspension, and inoculating the bacterial suspension to a herbal tea residue culture medium for fermentation; the herb tea residue comprises flos Plumeriae Acutifoliae, flos Lonicerae, flos Chrysanthemi, herba mesonae chinensis, Prunellae Spica, Glycyrrhrizae radix and folium Microcorii Paniculati.
2. The method as claimed in claim 1, further comprising activating and passaging Aspergillus niger in advance, wherein the activating step comprises inoculating Aspergillus niger to PDA solid medium culture.
3. The method according to claim 2, wherein the fermentation conditions are 50-90% humidity and 28-37 ℃ for 24-240 h.
4. The method of claim 1, wherein said bacterial suspension is at a concentration of 105～109cfu/mL, preferably at a bacterial suspension concentration of 2X 107cfu/mL。
5. The method of claim 1, wherein the aspergillus niger inoculum size is 5% to 25%.
6. The method according to claim 1, wherein the fermentation temperature is 28 ℃ to 37 ℃ and the fermentation time is 1 to 10 days.
7. The method according to claim 1, wherein the water content during fermentation is 65-85%, and the pH of the soaking solution is 5-10.
8. The method of claim 1, wherein the herbal tea pomace medium comprises: 1-5 parts of cold tea residue powder, 0.04-0.20 part of ammonium sulfate, 0-0.1 part of glucose, 0.005-0.02 part of monopotassium phosphate, 0.002-0.020 part of dipotassium hydrogen phosphate and 5.5-8.5 parts of water.
9. The method of claim 1, wherein the herbal tea pomace medium comprises: 2 parts of cold tea residue powder, 0.08 part of ammonium sulfate, 0.04 part of glucose, 0.01 part of monopotassium phosphate, 0.008 part of dipotassium hydrogen phosphate and 8 parts of water.
10. The method according to claim 8 or 9, wherein the cold tea leaf powder is: mixing flos Plumeriae Acutifoliae, flos Lonicerae, flos Chrysanthemi, herba mesonae chinensis, Prunellae Spica, Glycyrrhrizae radix and folium Microcorii Paniculati, and decocting to obtain herbal tea residue.
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| CN113061537A (en) * | 2021-04-02 | 2021-07-02 | 深高蓝德环保科技集团股份有限公司 | Method for preparing aspergillus niger microbial inoculum |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113061537A (en) * | 2021-04-02 | 2021-07-02 | 深高蓝德环保科技集团股份有限公司 | Method for preparing aspergillus niger microbial inoculum |
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