CN102321540A - Preparation method of lactic acid Amphibacillus fermentation liquor, and preparation method of lactic acid Amphibacillus fermentation liquor composite antiseptic by using fermentation liquor thereof - Google Patents
Preparation method of lactic acid Amphibacillus fermentation liquor, and preparation method of lactic acid Amphibacillus fermentation liquor composite antiseptic by using fermentation liquor thereof Download PDFInfo
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 166
- 239000004310 lactic acid Substances 0.000 title claims abstract description 83
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 83
- 238000002360 preparation method Methods 0.000 title claims abstract description 52
- 238000000855 fermentation Methods 0.000 title claims abstract description 23
- 230000004151 fermentation Effects 0.000 title claims abstract description 23
- 230000002421 anti-septic effect Effects 0.000 title abstract 7
- 241001147782 Amphibacillus Species 0.000 title abstract 5
- 239000002131 composite material Substances 0.000 title abstract 5
- 239000007788 liquid Substances 0.000 claims abstract description 136
- 241000894006 Bacteria Species 0.000 claims abstract description 74
- 238000000034 method Methods 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 31
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 claims abstract description 20
- 229920001661 Chitosan Polymers 0.000 claims abstract description 11
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 claims abstract description 11
- NVNLLIYOARQCIX-MSHCCFNRSA-N Nisin Chemical compound N1C(=O)[C@@H](CC(C)C)NC(=O)C(=C)NC(=O)[C@@H]([C@H](C)CC)NC(=O)[C@@H](NC(=O)C(=C/C)/NC(=O)[C@H](N)[C@H](C)CC)CSC[C@@H]1C(=O)N[C@@H]1C(=O)N2CCC[C@@H]2C(=O)NCC(=O)N[C@@H](C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(NCC(=O)N[C@H](C)C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCSC)C(=O)NCC(=O)N[C@H](CS[C@@H]2C)C(=O)N[C@H](CC(N)=O)C(=O)N[C@H](CCSC)C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(N[C@H](C)C(=O)N[C@@H]3C(=O)N[C@@H](C(N[C@H](CC=4NC=NC=4)C(=O)N[C@H](CS[C@@H]3C)C(=O)N[C@H](CO)C(=O)N[C@H]([C@H](C)CC)C(=O)N[C@H](CC=3NC=NC=3)C(=O)N[C@H](C(C)C)C(=O)NC(=C)C(=O)N[C@H](CCCCN)C(O)=O)=O)CS[C@@H]2C)=O)=O)CS[C@@H]1C NVNLLIYOARQCIX-MSHCCFNRSA-N 0.000 claims abstract description 10
- 108010053775 Nisin Proteins 0.000 claims abstract description 10
- NCXMLFZGDNKEPB-FFPOYIOWSA-N natamycin Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C[C@@H](C)OC(=O)/C=C/[C@H]2O[C@@H]2C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 NCXMLFZGDNKEPB-FFPOYIOWSA-N 0.000 claims abstract description 10
- 229960003255 natamycin Drugs 0.000 claims abstract description 10
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- 235000010297 nisin Nutrition 0.000 claims abstract description 10
- 235000019260 propionic acid Nutrition 0.000 claims abstract description 10
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 claims abstract description 10
- 230000004913 activation Effects 0.000 claims description 33
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 27
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 23
- 150000001875 compounds Chemical class 0.000 claims description 22
- 239000003755 preservative agent Substances 0.000 claims description 22
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- 241000235017 Zygosaccharomyces Species 0.000 claims description 14
- 239000002253 acid Substances 0.000 claims description 14
- 125000002723 alicyclic group Chemical group 0.000 claims description 14
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- 239000011259 mixed solution Substances 0.000 claims description 12
- LXAHHHIGZXPRKQ-UHFFFAOYSA-N 5-fluoro-2-methylpyridine Chemical compound CC1=CC=C(F)C=N1 LXAHHHIGZXPRKQ-UHFFFAOYSA-N 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 9
- 229940041514 candida albicans extract Drugs 0.000 claims description 8
- JEIPFZHSYJVQDO-UHFFFAOYSA-N ferric oxide Chemical compound O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 claims description 8
- 239000012138 yeast extract Substances 0.000 claims description 8
- 239000000284 extract Substances 0.000 claims description 7
- 239000002609 medium Substances 0.000 claims description 7
- 241001037822 Bacillus bacterium Species 0.000 claims description 6
- 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 5
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 3
- 229910019142 PO4 Inorganic materials 0.000 claims description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 3
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- 239000006071 cream Substances 0.000 claims description 3
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims description 3
- 239000008101 lactose Substances 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 3
- 239000010452 phosphate Substances 0.000 claims description 3
- 239000002953 phosphate buffered saline Substances 0.000 claims description 3
- 229910052700 potassium Inorganic materials 0.000 claims description 3
- 239000011591 potassium Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 3
- 229960001763 zinc sulfate Drugs 0.000 claims description 3
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 3
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical class CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 2
- 229940066779 peptones Drugs 0.000 claims description 2
- 241000194035 Lactococcus lactis Species 0.000 claims 4
- 235000013305 food Nutrition 0.000 abstract description 19
- 238000004519 manufacturing process Methods 0.000 abstract description 16
- 238000005516 engineering process Methods 0.000 abstract description 7
- GVJHHUAWPYXKBD-UHFFFAOYSA-N d-alpha-tocopherol Natural products OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 abstract description 3
- 235000017166 Bambusa arundinacea Nutrition 0.000 abstract description 2
- 235000017491 Bambusa tulda Nutrition 0.000 abstract description 2
- 235000015334 Phyllostachys viridis Nutrition 0.000 abstract description 2
- 239000011425 bamboo Substances 0.000 abstract description 2
- 241001330002 Bambuseae Species 0.000 abstract 1
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 abstract 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 abstract 1
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- 235000010298 natamycin Nutrition 0.000 abstract 1
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- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 description 2
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- 241000228153 Penicillium citrinum Species 0.000 description 2
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- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a preparation method of lactic acid Amphibacillus fermentation liquor, and a preparation method of a lactic acid Amphibacillus fermentation liquor composite antiseptic by using the fermentation liquor thereof. The method of the invention aims at solving the problems of present complex preparation technology of lactic acid Amphibacillus as well as high production cost and low output existed in antiseptic preparation. The preparation method for the fermentation liquor comprises the following steps: activating bacteria liquid; extracting thallus from the activated bacteria liquid of four types bacterium; mixing the thallus, inoculating into a fermentation cylinder for fermenting to obtain the fermentation liquor; culturing the fermentation liquor to finish the preparation of the fermentation liquor. The method for preparing the composite antiseptic comprises the following steps: mixing nisin, natamycin, propionic acid, bamboo leaf antioxidant, chitosan, disodium ethylene diamine tetraacetate, ethanol, tocopherol and activated water; mixing the mixed liquor and the fermentation liquor to react to finish the preparation of the composite antiseptic. The method of the invention has the advantages of simple preparation technology, low cost and high output. The composite antiseptic is used for food antiseptic.
Description
Technical field
The present invention relates to the preparation method of the two bud fermented liquids of a kind of lactic acid and utilize the two bud fermented liquids of this lactic acid to prepare the method for the two bud fermented liquid compound preservatives of lactic acid.
Background technology
The two bud bacterium of lactic acid are biological food sanitass, and its bactericidal ability is extensive, and effect is fine.Antifungal mechanism is to set up enzyme biosynthesizing and active adjusting pattern; On the microbial cell film, form micropore, cause membrane permeability increase, energy system to destroy and food toxin destruction, guarantee not produce sensibiligen in the food; Thereby can kill the mikrobe in the food rapidly; And a small amount of mikrobe in the food suppressed, avoid its breeding and water-soluble sugar protein immunization to react, thereby reach the purpose of anti-corrosive fresh-keeping.
At present, the complicated process of preparation of the two bud bacterium of lactic acid needs through Technologies such as a large amount of blowdown reaction treatment in the production, and often uses the chemosynthesis raw material, is prone to produce peculiar smell.There is production cost high (up to 25000 yuan/ton) in the preparation of its sanitas, yields poorly, and quantitatively serve as to produce to adjust promptly with investment cost, invest 1,000 ten thousand yuan, only can build 5000 tons/year production line.
Summary of the invention
The present invention is the complicated process of preparation that will solve the two bud bacterium of existing lactic acid; There is the problem that production cost is high and yield poorly in the preparation of its sanitas, and the preparation method of the two bud fermented liquids of lactic acid is provided and utilizes this fermented liquid to prepare the method for the two bud fermented liquid compound preservatives of lactic acid.
The preparation method of the two bud fermented liquids of lactic acid of the present invention carries out: one, respectively lactococcus lactis ssp, thermophilic alicyclic acid genus bacillus, spheroblast spore bacillus and two gemma zygosaccharomyceses are carried out activation, get the activation bacterium liquid of four kinds of bacterium according to the following steps; Two, from the activation bacterium liquid of four kinds of bacterium, extract the thalline of four kinds of bacterium; Three, the thalline of four kinds of bacterium that step 2 obtained by thalline quantity than 1: 1: 1: 1 mixes; Be inoculated in then in the fermentor tank that fermention medium is housed; Inoculum size is 8%～12%; At leavening temperature is that 29～31 ℃, shaking table speed are condition bottom fermentation 45～50h of 100～150r/min, fermented liquid; Four, fermented liquid is placed on the shaking table in 100～150r/min cultivate 45～50h, can accomplish the preparation of the two bud fermented liquids of lactic acid.
Utilize the two bud fermented liquids of above-mentioned lactic acid to prepare the method for the two bud fermented liquid compound preservatives of lactic acid; Carry out according to the following steps: one, by volume umber is that 75% ethanol, 0.2～0.6 part Viteolin and 75～85 parts activated water mix with 1～3 part nisin, 1～5 part tennecetin, 1～5 part propionic acid, 2～8 parts AOB, 2～5 parts chitosan, 2～4 parts EDTA Disodium, 0.2～0.8 part concentration of volume percent, obtains mixed solution; Two, by volume per-cent mixes 75～80 parts mixed solution and 20～25 parts the two bud fermented liquids of lactic acid, reacts 1～3h down at 45～55 ℃, can accomplish the preparation of the two bud fermented liquid compound preservatives of lactic acid.
The preparation technology of the two bud fermented liquids of lactic acid of the present invention is simple, need not pass through a large amount of blowdown reaction treatment, does not have the chemosynthesis raw material in process of production; A small amount of substratum residue is arranged during the fermentation, and residue is 0.75% (is the 100kg fermented liquid, the 0.75kg residue is arranged) of fermented liquid; Do not produce the three wastes, not blowdown, free from extraneous odour in process of production; Be closed low temperature fermentation and preparation in the various productions jars, hygienic condition are clean and tidy, the pollution-free source of room air; The purge tank water reclaims, and discharging is zero.
The cost that the inventive method prepares the two bud fermented liquid compound preservatives of lactic acid is low, is 15000 yuan/ton; Output is high, invests 1,000 ten thousand yuan, can build 6000 tons/year production line.
Embodiment
Technical scheme of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: the preparation method of the two bud fermented liquids of this embodiment lactic acid; Carry out according to the following steps: one, respectively lactococcus lactis ssp, thermophilic alicyclic acid genus bacillus, spheroblast spore bacillus and two gemma zygosaccharomyceses are carried out activation, get the activation bacterium liquid of four kinds of bacterium; Two, from the activation bacterium liquid of four kinds of bacterium, extract the thalline of four kinds of bacterium; Three, the thalline of four kinds of bacterium that step 2 obtained by thalline quantity than 1: 1: 1: 1 mixes; Be inoculated in then in the fermentor tank that fermention medium is housed; Inoculum size is 8%～12% (volume); At leavening temperature is that 29～31 ℃, shaking table speed are condition bottom fermentation 45～50h of 100～150r/min, fermented liquid; Four, fermented liquid is placed on the shaking table in 100～150r/min cultivate 45～50h, can accomplish the preparation of the two bud fermented liquids of lactic acid.
The preparation technology of the two bud fermented liquids of this embodiment lactic acid is simple, need not pass through a large amount of blowdown reaction treatment, does not have the chemosynthesis raw material in process of production; A small amount of substratum residue is arranged during the fermentation, and residue is 0.75% (is the 100kg fermented liquid, the 0.75kg residue is arranged) of fermented liquid; Do not produce the three wastes, not blowdown, free from extraneous odour in process of production; Be closed low temperature fermentation and preparation in the various productions jars, hygienic condition are clean and tidy, the pollution-free source of room air; The purge tank water reclaims, and discharging is zero.
The two bud fermented liquids of the lactic acid of this embodiment preparation are orange-yellow liquid, have a little leaf of bamboo pyramid-shaped dumpling fragrance.
The said lactococcus lactis ssp of this embodiment step 1 is bought from Institute of Microorganism, Academia Sinica common micro-organisms culture presevation administrative center, and culture presevation is numbered ACCC 10535r, and preservation date is June 1 calendar year 2001.
The said thermophilic alicyclic acid genus bacillus of this embodiment step 1 is bought from Institute of Microorganism, Academia Sinica common micro-organisms culture presevation administrative center, and culture presevation is numbered ACCC 1022lr, and preservation date is on June 8th, 2006.
The said spheroblast spore of this embodiment step 1 bacillus is bought from Shanghai Inst. of Biochemistry, Chinese Academy of Sciences, and culture presevation is numbered 1.1202, and preservation date is on May 9th, 2008.
The said two spore zygosaccharomyceses of this embodiment step 1 are bought from Chinese Guangdong province institute of microbiology, and culture presevation is numbered 2.1529, and preservation date is on June 5th, 2006.
The two bud fermented liquids of the lactic acid of this embodiment preparation are carried out toxicity to be detected: the searching (No. 2011020) of Shandong disease control poison, examining report CNSSL0119 (bw).The two bud fermented liquids of lactic acid, rat (magnetic, male) acute oral toxicological test MTD＞10000mg/kg (bw); In 14 days observation period, obvious pathological change is not seen in the sick substantially inspection of animal, does not see tangible acute poisoning reaction, does not also have animal dead, puts to death animal when experiment finishes, and carries out gross anatomy, and each internal organs is not seen obviously unusual.The result shows that this fermented liquid belongs to nontoxic level, quality standard: accord with Q/LYC01-2010.
The two bud fermented liquids of the lactic acid of this embodiment preparation carry out Oranoleptic indicator's detection, and the result is as shown in table 1:
Table 1
The two bud fermented liquids of the lactic acid of this embodiment preparation carry out physical and chemical index and detect, and the result is as shown in table 2:
Table 2
The two bud fermented liquids of the lactic acid of this embodiment preparation carry out the pollutent index and detect, and the result is as shown in table 3:
Table 3
The two bud fermented liquids of the lactic acid of this embodiment preparation carry out microbiological indicator and detect, and the result is as shown in table 4:
Table 4
The two bud fermented liquids of the lactic acid of this embodiment are added to play the effect of killing and suppressing yeast, mould, bacterium in the food.In use, can directly in food, add the two bud fermented liquids of lactic acid, also can the spray-dried pulvis of processing of fermented liquid be joined in the food.
The two bud fermented liquids of this embodiment gained lactic acid are safe and harmless to human body; In digestive tube, be degraded to the normal composition of food, do not influence the effect of medicinal microbiotic, the bactericidal ability is strong; Has the very strong effect of killing and suppress yeast, mould, bacterium (inhibited equally) to fungi; Test of many times shows through China National Academy of Food & Fermentation Industries, under addition and the identical situation of experiment condition, black mold, Penicillium citrinum, yeast saccharomyces cerevisiae, intestinal bacteria, subtilis, streptococcus aureus etc. is tried bacterium; The fungistatic effect of the two bud bacterium of lactic acid generally is higher than POTASSIUM SORBATE GRANULAR WHITE; Especially to causing the yeast of food spoilage, mould, bacteriological action is the strongest.Generally speaking; The mass percent concentration that the two bud fermented liquids of the lactic acid of this embodiment preparation add in the food is 0.1%～0.15%; The virtual mass percentage concentration of its mould fungus inhibition is 0.004%～0.08%, and the virtual mass percentage concentration that suppresses bacterium is 0.0012%～0.15%.
It is 0.3% can kill following Pseudomonas that the two bud fermented liquids of this embodiment lactic acid add mass percent concentration in the food to, and it is 0.1% can suppress following Pseudomonas that the two bud fermented liquids of lactic acid add mass percent concentration in the food to: Rhodopseudomonas, micrococcus sp, bacillus, Vibrio; Halobacterium, mould, lactobacillus, black mold, flavus; Penicillium citrinum, intestinal bacteria, proteus vulgaris, lactobacterium casei, Lactobacillus plantarum; Aerobic spore-bearing bacilli, Gram-negative bacteria, gram-positive microorganism, clostridium botulinum; Shigellae, Salmonellas, streptococcus aureus, Hemolytic streptococcus etc.; It is thus clear that the scope of restraining fungi of the two bud fermented liquids of this embodiment gained lactic acid is wide.
The two bud fermented liquids of this embodiment gained lactic acid are stable to the acid-basicity of food, adapt to the food of various potential of hydrogen, like pork, veal prods, fishery products; Fish product, pickling sauerkraut, pickles, the Tian jin cabbage pickled in sweet and sour of not pickling; Salting cucumber, Radix Raphani etc., jam product (soya sauce, northeast salty sauce, farm household sauce, broad bean paste etc.), greengrocery; Seasonings (soy sauce, light soy sauce, dark soy sauce, cooking wine, butter, rice wine etc.), dried aromatizing agent class, candied fruit filling material; Drink class, soybean product, metric system category and starch cake class, the mass percent concentration that in milk preparation, adds the two bud fermented liquids of lactic acid must not surpass 1%; It is thus clear that the usage quantity of the two bud fermented liquids of this embodiment gained lactic acid is little, applied range.
Embodiment two: what this embodiment and embodiment one were different is: respectively lactococcus lactis ssp, thermophilic alicyclic acid genus bacillus, spheroblast spore bacillus and two gemma zygosaccharomyceses being carried out the activatory method in the step 1 is: the lactococcus lactis ssp bacterium liquid that with concentration is 0.05～0.15g/kg is inoculated in the culture medium A by the inoculum size of 8%～12% (volume); The thermophilic alicyclic acid genus bacillus bacterium liquid that with concentration is 0.1～0.2g/kg is inoculated among the substratum B by the inoculum size of 8%～12% (volume); The spheroblast spore bacillus bacterium liquid that with concentration is 0.05～0.15g/kg is inoculated in the culture medium C by the inoculum size of 8%～12% (volume); The two gemma zygosaccharomyces bacterium liquid that with concentration are 0.2～0.4g/kg are inoculated among the substratum D by the inoculum size of 8%～12% (volume); In 28～32 ℃ of constant incubators, cultivate 45～50h then, promptly accomplish activation; Wherein every 200mL culture medium A is made up of the zero(ppm) water of 2g glucose, 1g Carnis Bovis seu Bubali cream, 1g lactose, 2g peptone, 1g yeast extract and surplus; Every 5000mL substratum B is made up of 2000mL solution a, 2000mL solution b and 1000mL zero(ppm) water; Said 2000mL solution a is made up of the zero(ppm) water of 6g potassium primary phosphate, 4g yeast extract paste and surplus, and said 2000mL solution b is made up of the zero(ppm) water of 0.2g zinc sulfate, 0.4g NSC 51149,0.6g boric acid, 0.06g Sodium orthomolybdate and surplus; Every 1000mL culture medium C is made up of the zero(ppm) water of the smart powder of 10g cottonseed, 0.6g beef extract and surplus; Substratum D is a wort.Other is identical with embodiment one.
Solution a and solution b among the said substratum B of this embodiment sterilize separately.
Embodiment three: what this embodiment was different with embodiment one or two is: the method for from the activation bacterium liquid of four kinds of bacterium, extracting the thalline of four kinds of bacterium in the step 2 is: the activation bacterium liquid of four kinds of bacterium is centrifugal respectively; Lactococcus lactis ssp activation bacterium liquid, thermophilic alicyclic acid genus bacillus activation bacterium liquid and spheroblast spore bacillus activation bacterium liquid are in the centrifugal 25～35min of 2500～3500r/min; Two gemma zygosaccharomyces activation bacterium liquid are in the centrifugal 10～20min of 2500～3500r/min; Collect four kinds of thalline then respectively; Adopting 0.4mol/L, pH value again is four kinds of thalline of 7.5 phosphate buffered saline buffer washing 3～5 times, and centrifugal collection thalline promptly obtains the thalline of four kinds of bacterium.Other is identical with embodiment one or two.
Embodiment four: what this embodiment was different with one of embodiment one to three is: inoculum size is 8% in the step 3.Other is identical with one of embodiment one to three.
Embodiment five: what this embodiment was different with one of embodiment one to three is: inoculum size is 12% in the step 3.Other is identical with one of embodiment one to three.
Embodiment six: what this embodiment was different with one of embodiment one to three is: inoculum size is 9%～11% in the step 3.Other is identical with one of embodiment one to three.
Embodiment seven: what this embodiment was different with one of embodiment one to three is: inoculum size is 10% in the step 3.Other is identical with one of embodiment one to three.
Embodiment eight: what this embodiment was different with one of embodiment one to seven is: leavening temperature is 29 ℃ in the step 3.Other is identical with one of embodiment one to seven.
Embodiment nine: what this embodiment was different with one of embodiment one to seven is: leavening temperature is 31 ℃ in the step 3.Other is identical with one of embodiment one to seven.
Embodiment ten: what this embodiment was different with one of embodiment one to seven is: leavening temperature is 30 ℃ in the step 3.Other is identical with one of embodiment one to seven.
Embodiment 11: what this embodiment was different with one of embodiment one to ten is: shaking table speed is 110～140r/min in the step 3.Other is identical with one of embodiment one to ten.
Embodiment 12: what this embodiment was different with one of embodiment one to ten is: shaking table speed is 120r/min in the step 3.Other is identical with one of embodiment one to ten.
Embodiment 13: what this embodiment was different with one of embodiment one to 12 is: 46～49h ferments in the step 3.Other is identical with one of embodiment one to 12.
Embodiment 14: what this embodiment was different with one of embodiment one to 12 is: 48h ferments in the step 3.Other is identical with one of embodiment one to 12.
Embodiment 15: what this embodiment was different with one of embodiment one to 14 is: every 2000mL fermention medium is by 20g glucose, 2g potassium hydrogenphosphate, 4g NH in the step 3
4NO
3, 0.4g MgSO
47H
2O, 0.01g MgSO
4, 0.01g FeSO
47H
2O, 0.2g CaCl
22H
2The zero(ppm) water of O, 6g yeast extract paste, the many peptones of 0.6g and surplus is formed.Other is identical with one of embodiment one to 14.
After the sterilization of the said fermention medium of this embodiment, be adjusted to pH value to 9.7 with the half yellow soda ash sterile solution of 1mol/L.
Embodiment 16: what this embodiment was different with one of embodiment one to 15 is: in the step 4 fermented liquid is placed on the shaking table in 110～140r/min cultivation, 46～49h.Other is identical with one of embodiment one to 15.
Embodiment 17: what this embodiment was different with one of embodiment one to 15 is: in the step 4 fermented liquid is placed on the shaking table in 120r/min cultivation 48h.Other is identical with one of embodiment one to 15.
Embodiment 18: the preparation method of the two bud fermented liquids of this embodiment lactic acid; Carry out according to the following steps: one, respectively lactococcus lactis ssp, thermophilic alicyclic acid genus bacillus, spheroblast spore bacillus and two gemma zygosaccharomyceses are carried out activation, get the activation bacterium liquid of four kinds of bacterium; Two, from the activation bacterium liquid of four kinds of bacterium, extract the thalline of four kinds of bacterium; Three, the thalline of four kinds of bacterium that step 2 obtained by thalline quantity than 1: 1: 1: 1 mixes; Be inoculated in then in the fermentor tank that fermention medium is housed; Inoculum size is 10% (volume), is that 30 ℃, shaking table speed are the condition bottom fermentation 48h of 120r/min at leavening temperature, fermented liquid; Four, fermented liquid is placed on the shaking table in 120r/min cultivate 48h, can accomplish the preparation of the two bud fermented liquids of lactic acid.
Respectively lactococcus lactis ssp, thermophilic alicyclic acid genus bacillus, spheroblast spore bacillus and two gemma zygosaccharomyceses being carried out the activatory method in this embodiment step 1 is: the lactococcus lactis ssp bacterium liquid that with concentration is 0.1g/kg is inoculated in the culture medium A by the inoculum size of 10% (volume); The thermophilic alicyclic acid genus bacillus bacterium liquid that with concentration is 0.15g/kg is inoculated among the substratum B by the inoculum size of 10% (volume); The spheroblast spore bacillus bacterium liquid that with concentration is 0.1g/kg is inoculated in the culture medium C by the inoculum size of 10% (volume); The two gemma zygosaccharomyces bacterium liquid that with concentration are 0.3g/kg are inoculated among the substratum D by the inoculum size of 10% (volume); In 30 ℃ of constant incubators, cultivate 48h then, promptly accomplish activation; Wherein every 200mL culture medium A is made up of the zero(ppm) water of 2g glucose, 1g Carnis Bovis seu Bubali cream, 1g lactose, 2g peptone, 1g yeast extract and surplus; Every 5000mL substratum B is made up of 2000mL solution a, 2000mL solution b and 1000mL zero(ppm) water; Said 2000mL solution a is made up of the zero(ppm) water of 6g potassium primary phosphate, 4g yeast extract paste and surplus, and said 2000mL solution b is made up of the zero(ppm) water of 0.2g zinc sulfate, 0.4g NSC 51149,0.6g boric acid, 0.06g Sodium orthomolybdate and surplus; Every 1000mL culture medium C is made up of the zero(ppm) water of the smart powder of 10g cottonseed, 0.6g beef extract and surplus; Substratum D is a wort.
The method of from the activation bacterium liquid of four kinds of bacterium, extracting the thalline of four kinds of bacterium in this embodiment step 2 is: the activation bacterium liquid of four kinds of bacterium is centrifugal respectively; Lactococcus lactis ssp activation bacterium liquid, thermophilic alicyclic acid genus bacillus activation bacterium liquid and spheroblast spore bacillus activation bacterium liquid are in the centrifugal 30min of 3000r/min; Two gemma zygosaccharomyces activation bacterium liquid are in the centrifugal 15min of 3000r/min; Collect four kinds of thalline then respectively; Adopting 0.4mol/L, pH value again is four kinds of thalline of 7.5 phosphate buffered saline buffer washing 3 times, and centrifugal collection thalline promptly obtains the thalline of four kinds of bacterium.
The preparation technology of the two bud fermented liquids of this embodiment lactic acid is simple, need not pass through a large amount of blowdown reaction treatment, does not have the chemosynthesis raw material in process of production; A small amount of substratum residue is arranged during the fermentation, and residue is 0.75% (is the 100kg fermented liquid, the 0.75kg residue is arranged) of fermented liquid; Do not produce the three wastes, not blowdown, free from extraneous odour in process of production; Be closed low temperature fermentation and preparation in the various productions jars, hygienic condition are clean and tidy, the pollution-free source of room air; The purge tank water reclaims, and discharging is zero.
Embodiment 19: this embodiment utilizes the two bud fermented liquids of lactic acid to prepare the method for the two bud fermented liquid compound preservatives of lactic acid; Carry out according to the following steps: one, by volume umber is that 75% ethanol, 0.2～0.6 part Viteolin and 75～85 parts activated water mix with 1～3 part nisin, 1～5 part tennecetin, 1～5 part propionic acid, 2～8 parts AOB, 2～5 parts chitosan, 2～4 parts EDTA Disodium, 0.2～0.8 part concentration of volume percent, obtains mixed solution; Two, by volume per-cent mixes 75～80 parts mixed solution and 20～25 parts the two bud fermented liquids of lactic acid, reacts 1～3h down at 45～55 ℃, can accomplish the preparation of the two bud fermented liquid compound preservatives of lactic acid.
The compound preservative of this embodiment preparation directly adds in the food in use, and addition is 0.1g/kg.
The said activated water of step 1 is to be made by the two-pass reverse osmosis water treating equipment.
Ethanol is the auxiliary agent of in GB2760-2007, having stipulated to allow in the food-processing use.
Chitosan is the thickening material of in the GB2760-2007 function, stipulating as the use of fruit glaze agent, fruit glaze agent.
Viteolin uses as inhibitor, is the inhibitor of in the GB2760-2007 function, stipulating.
The two bud fermented liquid compound preservatives of the lactic acid of this embodiment preparation, it is higher 10 times than effective quality guaranteed period of Chemical Preservation Sodium Benzoate in food, to add same dosage, higher 6 times than effective quality guaranteed period of POTASSIUM SORBATE GRANULAR WHITE.
The two bud fermented liquid compound preservatives of the lactic acid of this embodiment preparation are carried out toxicity to be detected: Shandong disease control poison searching (No. 2011020) examining report CNASL0119.The two bud fermented liquid compound preservatives of lactic acid, rat acute per os toxicity test MTD＞10000mL/kg (bw).In 14 days observation period, animal disease inspection does not substantially see obvious pathological change (do not see tangible acute poisoning reaction, different no animal dead is put to death animal during off-test, carry out gross anatomy, and each device is dirty not to be seen obviously unusually).The result shows that this compound preservative belongs to actual nontoxic level.
The CNS of nisin number is 17.019; The CNS of tennecetin number is 10.030, and the CNS of propionic acid number is 17.029,17.005 or 17.006, and the CNS of AOB number is 04.019; The CNS of chitosan number is 20.026; EDTA Disodium (EDTA) is a C.1 process for processing auxiliary agent of GB2760-2007 standardization appendix C table, and ethanol is C.1 process for processing auxiliary agent of GB2760-2007 standardization appendix C table, and the CNS of Viteolin (Dl-a-Viteolin, vitamin E) number 04.016.
Embodiment 20: what this embodiment and embodiment 19 were different is: in the step 1 by volume umber be that 75% ethanol, 0.3～0.5 part Viteolin and 78～82 parts activated water mix with 2 parts nisin, 2～4 parts tennecetin, 2～4 parts propionic acid, 4～6 parts AOB, 3～4 parts chitosan, 3 parts EDTA Disodium, 0.4～0.6 part concentration of volume percent.Other is identical with embodiment 19.
Embodiment 21: what this embodiment and embodiment 19 were different is: in the step 1 by volume umber be that 75% ethanol, 0.4 part Viteolin and 80 parts activated water mix with 2 parts nisin, 3 parts tennecetin, 3 parts propionic acid, 5 parts AOB, 3.5 parts chitosan, 3 parts EDTA Disodium, 0.5 part concentration of volume percent.Other is identical with embodiment 19.
Embodiment 22: what this embodiment and embodiment 19 were different is: in the step 1 by volume umber be that 75% ethanol, 0.5 part Viteolin and 80 parts activated water mix with 2.5 parts nisin, 2 parts tennecetin, 3 parts propionic acid, 5 parts AOB, 3.5 parts chitosan, 3 parts EDTA Disodium, 0.5 part concentration of volume percent.Other is identical with embodiment 19.
Embodiment 23: what this embodiment was different with one of embodiment 19 to 22 is: in the step 2 by volume per-cent the two bud fermented liquids of lactic acid of 76～79 parts mixed solution and 21～24 parts are mixed.Other is identical with one of embodiment 19 to 22.
Embodiment 24: what this embodiment was different with one of embodiment 19 to 22 is: in the step 2 by volume per-cent the two bud fermented liquids of lactic acid of 78 parts mixed solution and 22 parts are mixed.Other is identical with one of embodiment 19 to 22.
Embodiment 25: what this embodiment was different with one of embodiment 19 to 22 is: in the step 2 by volume per-cent the two bud fermented liquids of lactic acid of 77 parts mixed solution and 23 parts are mixed.Other is identical with one of embodiment 19 to 22.
Embodiment 26: what this embodiment was different with one of embodiment 19 to 25 is: react 1h down at 45 ℃ in the step 2.Other is identical with one of embodiment 19 to 25.
Embodiment 27: what this embodiment was different with one of embodiment 19 to 25 is: react 3h down at 55 ℃ in the step 2.Other is identical with one of embodiment 19 to 25.
Embodiment 28: what this embodiment was different with one of embodiment 19 to 25 is: react 2h down at 50 ℃ in the step 2.Other is identical with one of embodiment 19 to 25.
Embodiment 29: this embodiment utilizes the two bud fermented liquids of lactic acid to prepare the method for the two bud fermented liquid compound preservatives of lactic acid; Carry out according to the following steps: one, by volume umber is that 75% ethanol, 0.5 part Viteolin and 80 parts activated water mix with 2.5 parts nisin, 2 parts tennecetin, 3 parts propionic acid, 5 parts AOB, 3.5 parts chitosan, 3 parts EDTA Disodium, 0.5 part concentration of volume percent, obtains mixed solution; Two, by volume per-cent mixes 78 parts mixed solution and 22 parts the two bud fermented liquids of lactic acid, reacts 2h down at 48 ℃, can accomplish the preparation of the two bud fermented liquid compound preservatives of lactic acid.
The method cost that this embodiment prepares the two bud fermented liquid compound preservatives of lactic acid is low, is 15000 yuan/ton; Output is high, invests 1,000 ten thousand yuan, can build 6000 tons/year production line.
Claims (10)
1. the preparation method of the two bud fermented liquids of lactic acid; The preparation method who it is characterized in that the two bud fermented liquids of lactic acid; Carry out according to the following steps: one, respectively lactococcus lactis ssp, thermophilic alicyclic acid genus bacillus, spheroblast spore bacillus and two gemma zygosaccharomyceses are carried out activation, get the activation bacterium liquid of four kinds of bacterium; Two, from the activation bacterium liquid of four kinds of bacterium, extract the thalline of four kinds of bacterium; Three, the thalline of four kinds of bacterium that step 2 obtained by thalline quantity than 1: 1: 1: 1 mixes; Be inoculated in then in the fermentor tank that fermention medium is housed; Inoculum size is 8%～12%; At leavening temperature is that 29～31 ℃, shaking table speed are condition bottom fermentation 45～50h of 100～150r/min, fermented liquid; Four, fermented liquid is placed on the shaking table in 100～150r/min cultivate 45～50h, can accomplish the preparation of the two bud fermented liquids of lactic acid.
2. the preparation method of the two bud fermented liquids of lactic acid according to claim 1; It is characterized in that respectively lactococcus lactis ssp, thermophilic alicyclic acid genus bacillus, spheroblast spore bacillus and two gemma zygosaccharomyceses being carried out the activatory method in the step 1 is: the lactococcus lactis ssp bacterium liquid that with concentration is 0.05～0.15g/kg is inoculated in the culture medium A by 8%～12% inoculum size; The thermophilic alicyclic acid genus bacillus bacterium liquid that with concentration is 0.1～0.2g/kg is inoculated among the substratum B by 8%～12% inoculum size; The spheroblast spore bacillus bacterium liquid that with concentration is 0.05～0.15g/kg is inoculated in the culture medium C by 8%～12% inoculum size; The two gemma zygosaccharomyces bacterium liquid that with concentration are 0.2～0.4g/kg are inoculated among the substratum D by 8%～12% inoculum size; In 28～32 ℃ of constant incubators, cultivate 45～50h then, promptly accomplish activation; Wherein every 200mL culture medium A is made up of the zero(ppm) water of 2g glucose, 1g Carnis Bovis seu Bubali cream, 1g lactose, 2g peptone, 1g yeast extract and surplus; Every 5000mL substratum B is made up of 2000mL solution a, 2000mL solution b and 1000mL zero(ppm) water; Said 2000mL solution a is made up of the zero(ppm) water of 6g potassium primary phosphate, 4g yeast extract paste and surplus, and said 2000mL solution b is made up of the zero(ppm) water of 0.2g zinc sulfate, 0.4g NSC 51149,0.6g boric acid, 0.06g Sodium orthomolybdate and surplus; Every 1000mL culture medium C is made up of the zero(ppm) water of the smart powder of 10g cottonseed, 0.6g beef extract and surplus; Substratum D is a wort.
3. the preparation method of the two bud fermented liquids of lactic acid according to claim 1 and 2; It is characterized in that the method for from the activation bacterium liquid of four kinds of bacterium, extracting the thalline of four kinds of bacterium in the step 2 is: the activation bacterium liquid of four kinds of bacterium is centrifugal respectively; Lactococcus lactis ssp activation bacterium liquid, thermophilic alicyclic acid genus bacillus activation bacterium liquid and spheroblast spore bacillus activation bacterium liquid are in the centrifugal 25～35min of 2500～3500r/min; Two gemma zygosaccharomyces activation bacterium liquid are in the centrifugal 10～20min of 2500～3500r/min; Collect four kinds of thalline then respectively; Adopting 0.4mol/L, pH value again is four kinds of thalline of 7.5 phosphate buffered saline buffer washing 3～5 times, and centrifugal collection thalline promptly obtains the thalline of four kinds of bacterium.
4. the preparation method of the two bud fermented liquids of lactic acid according to claim 3 is characterized in that inoculum size is 10% in the step 3.
5. the preparation method of the two bud fermented liquids of lactic acid according to claim 4 is characterized in that every 2000mL fermention medium is by 20g glucose, 2g potassium hydrogenphosphate, 4g NH in the step 3
4NO
3, 0.4g MgSO
47H
2O, 0.01gMgSO
4, 0.01gFeSO
47H
2O, 0.2g CaCl
22H
2The zero(ppm) water of O, 6g yeast extract paste, the many peptones of 0.6g and surplus is formed.
6. the preparation method of the two bud fermented liquids of lactic acid according to claim 5 is characterized in that in the step 4 fermented liquid being placed on the shaking table in 120r/min cultivation 48h.
7. the two bud fermented liquids of the lactic acid that utilizes claim 1 to prepare prepare the method for the two bud fermented liquid compound preservatives of lactic acid; It is characterized in that utilizing the two bud fermented liquids of lactic acid to prepare the method for the two bud fermented liquid compound preservatives of lactic acid; Carry out according to the following steps: one, by volume umber is that 75% ethanol, 0.2～0.6 part Viteolin and 75～85 parts activated water mix with 1～3 part nisin, 1～5 part tennecetin, 1～5 part propionic acid, 2～8 parts AOB, 2～5 parts chitosan, 2～4 parts EDTA Disodium, 0.2～0.8 part concentration of volume percent, obtains mixed solution; Two, by volume per-cent mixes 75～80 parts mixed solution and 20～25 parts the two bud fermented liquids of lactic acid, reacts 1～3h down at 45～55 ℃, can accomplish the preparation of the two bud fermented liquid compound preservatives of lactic acid.
8. the method for utilizing the two bud fermented liquids of lactic acid to prepare the two bud fermented liquid compound preservatives of lactic acid according to claim 7 is characterized in that in the step 1 that by volume umber is that 75% ethanol, 0.5 part Viteolin and 80 parts activated water mix with 2.5 parts nisin, 2 parts tennecetin, 3 parts propionic acid, 5 parts AOB, 3.5 parts chitosan, 3 parts EDTA Disodium, 0.5 part concentration of volume percent.
9. according to claim 7 or the 8 described methods of utilizing the two bud fermented liquids of lactic acid to prepare the two bud fermented liquid compound preservatives of lactic acid, it is characterized in that in the step 2 that by volume per-cent mixes 78 parts mixed solution and 22 parts the two bud fermented liquids of lactic acid.
10. the method for utilizing the two bud fermented liquids of lactic acid to prepare the two bud fermented liquid compound preservatives of lactic acid according to claim 9 is characterized in that reacting 2h down at 50 ℃ in the step 2.
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|---|---|---|---|---|
| CN102860457A (en) * | 2012-09-29 | 2013-01-09 | 上海韬鸿投资管理咨询有限公司 | Laxative detox nutrition powder |
| CN102860566A (en) * | 2012-09-29 | 2013-01-09 | 上海韬鸿投资管理咨询有限公司 | Biological food preservative |
| CN102972610A (en) * | 2012-11-27 | 2013-03-20 | 重庆市中药研究院 | Special composite preservative for walnut cake and application thereof |
| CN103355730A (en) * | 2013-08-06 | 2013-10-23 | 山东省蓝源生物工程有限公司 | Nisin composite biological preservative and preparation method thereof |
| CN103749663A (en) * | 2014-01-27 | 2014-04-30 | 四川赤健中药科技有限公司 | Fresh rhizoma gastrodiae preservative and preparation method thereof as well as fresh keeping method of fresh rhizoma gastrodiae |
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2011
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| MICHAEL E. STILES，ET AL.: "Biopreservation by lactic acid bacteria", 《ANTONIE VAN LEEUWENHOEK》 * |
| 潘恩霖: "微生物作为天然防腐剂的应用和展望", 《南平师专学报》 * |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102860457A (en) * | 2012-09-29 | 2013-01-09 | 上海韬鸿投资管理咨询有限公司 | Laxative detox nutrition powder |
| CN102860566A (en) * | 2012-09-29 | 2013-01-09 | 上海韬鸿投资管理咨询有限公司 | Biological food preservative |
| CN102860566B (en) * | 2012-09-29 | 2013-06-26 | 上海韬鸿投资管理咨询有限公司 | Biological food preservative |
| CN102972610A (en) * | 2012-11-27 | 2013-03-20 | 重庆市中药研究院 | Special composite preservative for walnut cake and application thereof |
| CN102972610B (en) * | 2012-11-27 | 2015-07-08 | 重庆市中药研究院 | Special composite preservative for walnut cake and application thereof |
| CN103355730A (en) * | 2013-08-06 | 2013-10-23 | 山东省蓝源生物工程有限公司 | Nisin composite biological preservative and preparation method thereof |
| CN103355730B (en) * | 2013-08-06 | 2015-07-15 | 山东省蓝源生物工程有限公司 | Nisin composite biological preservative and preparation method thereof |
| CN103749663A (en) * | 2014-01-27 | 2014-04-30 | 四川赤健中药科技有限公司 | Fresh rhizoma gastrodiae preservative and preparation method thereof as well as fresh keeping method of fresh rhizoma gastrodiae |
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| Publication number | Publication date |
|---|---|
| CN102321540B (en) | 2013-03-13 |
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