CN105950694B - Novel process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid - Google Patents
Novel process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid Download PDFInfo
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- 239000007788 liquid Substances 0.000 title claims abstract description 99
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 51
- 239000002921 fermentation waste Substances 0.000 title claims abstract description 43
- 150000004676 glycans Chemical class 0.000 title claims abstract description 33
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 33
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 33
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 33
- 238000000034 method Methods 0.000 title claims abstract description 26
- 241000209140 Triticum Species 0.000 title claims abstract description 21
- 235000021307 Triticum Nutrition 0.000 title claims abstract description 21
- 238000000855 fermentation Methods 0.000 claims abstract description 54
- 230000004151 fermentation Effects 0.000 claims abstract description 54
- 239000002699 waste material Substances 0.000 claims abstract description 33
- 241000193752 Bacillus circulans Species 0.000 claims abstract description 6
- 241000186605 Lactobacillus paracasei Species 0.000 claims abstract description 6
- 108091005804 Peptidases Proteins 0.000 claims abstract description 6
- 239000004365 Protease Substances 0.000 claims abstract description 6
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 6
- 239000002244 precipitate Substances 0.000 claims description 37
- 239000000243 solution Substances 0.000 claims description 31
- 238000001914 filtration Methods 0.000 claims description 25
- 239000011259 mixed solution Substances 0.000 claims description 23
- 230000001954 sterilising effect Effects 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 239000000843 powder Substances 0.000 claims description 17
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 15
- 239000000706 filtrate Substances 0.000 claims description 15
- 238000000926 separation method Methods 0.000 claims description 14
- 238000002156 mixing Methods 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 11
- 102000004190 Enzymes Human genes 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 10
- 229940088598 enzyme Drugs 0.000 claims description 10
- 229910052760 oxygen Inorganic materials 0.000 claims description 10
- 239000001301 oxygen Substances 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 10
- 239000012153 distilled water Substances 0.000 claims description 9
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 238000001816 cooling Methods 0.000 claims description 7
- 238000007710 freezing Methods 0.000 claims description 7
- 230000008014 freezing Effects 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 6
- 229920001661 Chitosan Polymers 0.000 claims description 5
- 241000186660 Lactobacillus Species 0.000 claims description 5
- 244000061456 Solanum tuberosum Species 0.000 claims description 5
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 5
- 102000004139 alpha-Amylases Human genes 0.000 claims description 5
- 108090000637 alpha-Amylases Proteins 0.000 claims description 5
- 229940024171 alpha-amylase Drugs 0.000 claims description 5
- 238000009835 boiling Methods 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 125000004122 cyclic group Chemical group 0.000 claims description 5
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 5
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 5
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 5
- 239000001963 growth medium Substances 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 230000000415 inactivating effect Effects 0.000 claims description 5
- 229940039696 lactobacillus Drugs 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- 235000006408 oxalic acid Nutrition 0.000 claims description 5
- 238000010298 pulverizing process Methods 0.000 claims description 5
- 238000007873 sieving Methods 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 5
- 238000000108 ultra-filtration Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- 235000015278 beef Nutrition 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 4
- 238000011218 seed culture Methods 0.000 claims description 4
- 230000001580 bacterial effect Effects 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 3
- 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 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 2
- 230000003698 anagen phase Effects 0.000 claims description 2
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 claims description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims description 2
- 108010009004 proteose-peptone Proteins 0.000 claims description 2
- 239000001632 sodium acetate Substances 0.000 claims description 2
- 235000017281 sodium acetate Nutrition 0.000 claims description 2
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 239000013543 active substance Substances 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 238000004064 recycling Methods 0.000 abstract description 2
- 230000007062 hydrolysis Effects 0.000 abstract 1
- 238000006460 hydrolysis reaction Methods 0.000 abstract 1
- 238000001556 precipitation Methods 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 238000011084 recovery Methods 0.000 description 8
- 239000003337 fertilizer Substances 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 6
- 244000005700 microbiome Species 0.000 description 5
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- 239000000463 material Substances 0.000 description 3
- 235000013379 molasses Nutrition 0.000 description 3
- 150000002894 organic compounds Chemical class 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 241000609240 Ambelania acida Species 0.000 description 2
- 240000003183 Manihot esculenta Species 0.000 description 2
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 241001148470 aerobic bacillus Species 0.000 description 2
- 239000010905 bagasse Substances 0.000 description 2
- 239000001506 calcium phosphate Substances 0.000 description 2
- 229910000389 calcium phosphate Inorganic materials 0.000 description 2
- 235000011010 calcium phosphates Nutrition 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 239000003895 organic fertilizer Substances 0.000 description 2
- 239000010802 sludge Substances 0.000 description 2
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 2
- 239000002351 wastewater Substances 0.000 description 2
- 241000228245 Aspergillus niger Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 1
- 241000221960 Neurospora Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 235000011941 Tilia x europaea Nutrition 0.000 description 1
- 241000223261 Trichoderma viride Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- DNZWLJIKNWYXJP-UHFFFAOYSA-N butan-1-ol;propan-2-one Chemical compound CC(C)=O.CCCCO DNZWLJIKNWYXJP-UHFFFAOYSA-N 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- FVSKHRXBFJPNKK-UHFFFAOYSA-N propionitrile Chemical compound CCC#N FVSKHRXBFJPNKK-UHFFFAOYSA-N 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
Abstract
The invention aims to provide a novel process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid, belonging to the field of waste recycling. The process uses molasses alcohol waste liquid as a raw material, sequentially enriches substances such as polysaccharide, active peptide and the like in the waste liquid through first-stage fermentation of bacillus circulans and second-stage fermentation of lactobacillus paracasei, then obtains the polysaccharide in a mode of respective alcohol precipitation, and obtains the active peptide in a mode of protease hydrolysis. The invention produces active substances by utilizing waste liquor of alcohol fermentation, can realize the reutilization of the waste, and has good environmental protection significance and economic value.
Description
Technical Field
The invention relates to a novel process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid, belonging to the field of waste recycling.
Background
In the process of sugar making and/or combined sugar making and alcohol making (making alcohol by utilizing molasses wastewater generated after sugar making), the largest pollutant generated is molasses alcohol fermentation waste liquid. The molasses alcohol fermentation waste liquid is high-COD waste water with strong corrosivity (acidity), and can cause serious pollution when directly discharged into the environment. The fermentation broth contains very little utilizable material and a large amount of impurities including pigments, colloids, cells themselves, unused medium, other unknown metabolites of the producing bacteria. The presence of these impurities makes it difficult to extract and purify the desired product from the fermentation waste liquid.
The method for treating the molasses alcohol fermentation waste liquid into fertilizer is researched, so that the harm is changed into the treasure. For example, chinese patent 90110379.9 discloses a method for manufacturing organic compound fertilizer, which comprises evaporating and concentrating molasses fermentation waste liquid, adding acid for digestion, and adding calcium phosphate and other aggregates, wherein no microorganism fermentation is used. However, since molasses fermentation waste liquid is acidic, and the pH value is lower after adding acid, aggregates such as calcium phosphate are easily acidified, so that phosphorus is lost, and fertility is affected.
Chinese patent 97112080.3 discloses a method for producing active organic compound fertilizer by using molasses alcohol waste liquid, which comprises mixing alcohol waste liquid, bagasse, filter mud and sludge, fermenting with natural aerobic bacteria carried by air neutralization material, and adding saprophytic microorganisms collected from the vicinity of production site for fermentation. The biggest problem of the method is that the quality of filter mud, sludge, aerobic bacteria and spoilage organisms is difficult to control, and unstable fermentation causes unstable quality of fertilizer products and is difficult to popularize industrially.
Chinese patent 01138281.3 discloses a method for producing organic compound fertilizer by using molasses alcohol lees (alcohol waste liquor), which comprises neutralizing with sodium hydroxide, evaporating and concentrating, adding lime for slaking, and mixing with fertilizer, wherein microorganism is used for fermentation. The method has no step of degrading other harmful wastes (such as alcohol and other volatile products) in the molasses alcohol fermentation waste liquid, directly evaporates and discharges into the atmosphere, and causes environmental pollution around a treatment plant.
Chinese patent 200310111476.4 discloses a method for producing organic fertilizer by fermenting a composition mainly comprising molasses alcohol waste liquid, bagasse and filter mud with Aspergillus niger, Trichoderma viride, Neurospora suberectus, Bacillus subtilis and cerevisiae Fermentum. Wherein, the adding amount of the animal organic fertilizer and the inorganic fertilizer is very small.
According to the research on butanol fermentation of ethanol fermentation waste liquid, the ethanol fermentation waste liquid is used as process water, a proper amount of corn flour or cassava is added to be used as a culture medium of acetone-butanol strains, acetone and butanol are produced through biological fermentation, waste materials are changed into valuable materials, the using amount of the corn flour and the cassava obtained through the propionitrile fermentation is saved, the cost is reduced, and the water consumption is also saved.
In the prior art, no report that wheat polysaccharide and active peptide can be co-produced from alcohol fermentation waste liquid is found. The invention aims to provide a process for carrying out resource treatment on waste liquid generated in the fermentation process of grain wine and simultaneously obtaining wheat polysaccharide and active peptide.
Disclosure of Invention
The invention aims to provide a novel process for coproducing wheat polysaccharide and active peptide by utilizing alcohol fermentation waste liquid. The invention produces active substances by utilizing waste liquor of alcohol fermentation, can realize the reutilization of the waste, and has good environmental protection significance and economic value.
The invention realizes the technical purpose through the following technical scheme:
novel process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid
1) Pretreatment of fermentation waste liquid: filtering the molasses alcohol waste liquid by using a coarse grating, filtering by using an ultrafiltration membrane with the diameter of 0.25 mu m, adding 0.1 percent of chitosan into the filtrate, strongly stirring, standing for 20-24h, carrying out ultrasonic treatment on the fermentation waste liquid for 5min, and filtering to obtain the pretreated molasses alcohol waste liquid;
2) primary fermentation of fermentation waste liquid: adding 0.5-2% of potato powder, 1-2% of yeast extract and 0.005% of magnesium sulfate into the pretreated molasses alcohol waste liquid, adjusting the pH value to 6.2-7.5 after mixing, sterilizing for 20min at 121 ℃, cooling the mixed solution to room temperature, inoculating 5-8% of cyclic bacillus seed liquid into the mixed solution, controlling the fermentation temperature to be 28-32 ℃, controlling the dissolved oxygen to be 50-70% for batch fermentation, sterilizing for 30-35 hours after fermentation, filtering, removing filter residues, centrifuging the filtrate to obtain a precipitate A and a separating liquid B, and continuing secondary fermentation of the centrifuged separating liquid;
3) two-stage fermentation of fermentation waste liquid: adding 0.5-2% of peptone powder, 1-2% of bran powder and 0.005% of monopotassium phosphate into the centrifuged separation solution, adjusting the pH value of the mixture to 5.2-6.5, sterilizing at 121 ℃ for 20min, cooling the mixed solution to room temperature, inoculating 8-10% of lactobacillus casei-like seed solution into the mixed solution, controlling the fermentation temperature to be 32-34 ℃, controlling the dissolved oxygen to be 50-70% for batch fermentation, sterilizing for 24-28 hours, filtering, removing filter residues, and centrifuging the filtrate to obtain a precipitate C and a separation solution D;
4) and (3) extracting polysaccharide: concentrating the separation liquid D to 1/6-1/8 of the original volume through a rotary evaporator, adding 0.1-0.3% of alpha-amylase, performing enzymolysis for 20-30 min under the assistance of ultrasonic waves at the temperature of 30 ℃ and at the pH value of 6.2, boiling to inactivate the enzyme after the enzymolysis is finished, centrifuging the enzymolysis liquid, adding 90% of ethanol water solution into the centrifuged separation liquid to obtain precipitate, and washing and drying the precipitate to obtain the wheat polysaccharide;
5) extraction of active peptide: mixing the precipitate A and the precipitate C, drying, pulverizing, sieving with 80 mesh sieve, adding 10 times of water, adjusting pH to 9.5-11.0 with sodium hydroxide, stirring in a constant temperature water bath at 60-75 deg.C for 1-2 hr, freeze centrifuging for 5min, adding oxalic acid into the supernatant to adjust pH to 4.2-4.8; freezing and centrifuging for 10min, removing supernatant, collecting precipitate, adding 6 times of distilled water into the precipitate, controlling the temperature of the solution at 35-40 deg.C, adding 0.05% protease composition, performing enzymatic hydrolysis for 1.5-3 hr, heating to 80 deg.C, and inactivating enzyme to obtain solution containing active peptide.
In the new process for co-producing the wheat polysaccharide and the active peptide by using the alcohol fermentation waste liquid, the bacillus circulans is used in the step 2Bacillus circulansIs preserved in China center for culture Collection of industrial microorganisms, and is numbered CICC 23053, and the composition of the seed culture medium comprises peptone 5.0g, beef extract 3.0g, NaCl5.0g, MnSO 4. H2O 5mg, distilled water 1.0L, and pH7.0.
The lactobacillus paracasei in the step 3Lactobacillus paracasei）Is preserved in China center for culture collection of industrial microorganisms with the number of CICC 20245, and the seed culture medium comprises casein peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, Tween 801.0 g, K2HPO42.0g，MgSO4.7H2O 0.2g，MnSO4.H2O0.05g, distilled water 1.0L, pH6.8.
In the new process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid, the lactobacillus paracasei and the bacillus circulans are addedThe access is in logarithmic growth phase. The number of bacterial colonies in the lactobacillus paracasei seed liquid is not less than 2 multiplied by 109The number of bacterial colonies in the bacillus circulans seed liquid is not less than 5 multiplied by 109One per ml.
Compared with the prior art, the invention has the following beneficial effects:
the method utilizes the molasses alcohol waste liquid as a raw material to finally prepare the polysaccharide and the active peptide, both have higher nutritive values, realize the effective utilization of waste resources, and have good environmental protection significance and economic value. Meanwhile, the production process is simple and easy to implement, strong in operability and good in application prospect. The recovery rate of the polysaccharide prepared by the process is 30-40%, and the recovery rate of the active peptide is 45-60%.
Detailed Description
The invention is further described below by means of specific examples.
Example 1
A new process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid comprises the following steps:
1) pretreatment of fermentation waste liquid: filtering the molasses alcohol waste liquid by using a coarse grating, filtering by using an ultrafiltration membrane with the diameter of 0.25 mu m, adding 0.1 percent of chitosan into the filtrate, strongly stirring, standing for 20 hours, carrying out ultrasonic treatment on the fermentation waste liquid for 5min, and filtering to obtain the pretreated molasses alcohol waste liquid;
2) primary fermentation of fermentation waste liquid: adding 0.5% of potato powder, 1% of yeast extract and 0.005% of magnesium sulfate into the pretreated molasses alcohol waste liquid, adjusting the pH value to 6.2 after mixing, sterilizing for 20min at 121 ℃, inoculating 5% of cyclic bacillus seed liquid into the mixed solution after the mixed solution is cooled to room temperature, controlling the fermentation temperature to 28 ℃, controlling the dissolved oxygen to be 50% for batch fermentation, sterilizing for 30 hours after fermentation, removing filter residues after filtration, centrifuging the filtrate to obtain a precipitate A and a separating liquid B, and continuously performing secondary fermentation on the centrifuged separating liquid;
3) two-stage fermentation of fermentation waste liquid: adding 0.5% of peptone powder, 1% of bran powder and 0.005% of monopotassium phosphate into the centrifuged separation solution, adjusting the pH value of the mixed solution to 5.2, sterilizing the mixed solution at 121 ℃ for 20min, cooling the mixed solution to room temperature, inoculating 8% of lactobacillus casei-like seed solution into the mixed solution, controlling the fermentation temperature to be 32 ℃, controlling the dissolved oxygen to be 50% for batch fermentation, sterilizing the mixed solution for 24 hours, filtering, removing filter residues, and centrifuging the filtrate to obtain a precipitate C and a separation solution D;
4) and (3) extracting polysaccharide: concentrating the separated liquid D to 1/6 of the original volume by a rotary evaporator, adding 0.1% alpha-amylase, performing enzymolysis for 20min under the assistance of ultrasonic waves at the temperature of 30 ℃ and the pH value of 6.2, boiling to inactivate the enzyme after the enzymolysis is finished, centrifuging the enzymolysis liquid, adding 90% ethanol water solution into the centrifuged separated liquid to obtain a precipitate, and washing and drying the precipitate to obtain the wheat polysaccharide;
5) extraction of active peptide: mixing the precipitate A and the precipitate C, drying, pulverizing, sieving with 80 mesh sieve, adding 10 times of water, adjusting pH to 9.5 with sodium hydroxide, stirring at 60 deg.C for 1 hr, freeze centrifuging for 5min, and adding oxalic acid into the supernatant to adjust pH to 4.2; freezing and centrifuging for 10min, removing supernatant, collecting precipitate, adding 6 times of distilled water, controlling solution temperature at 35 deg.C, adding 0.05% protease composition, performing enzymatic hydrolysis for 1.5 hr, heating to 80 deg.C, and inactivating enzyme to obtain solution containing active peptide.
The recovery rate of polysaccharide in the process is 30 percent, and the recovery rate of active peptide is 45 percent.
Example 2
A new process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid comprises the following steps:
1) pretreatment of fermentation waste liquid: filtering the molasses alcohol waste liquid by using a coarse grating, filtering by using an ultrafiltration membrane with the diameter of 0.25 mu m, adding 0.1 percent of chitosan into the filtrate, strongly stirring, standing for 22 hours, carrying out ultrasonic treatment on the fermented waste liquid for 5min, and filtering to obtain the pretreated molasses alcohol waste liquid;
2) primary fermentation of fermentation waste liquid: adding 1.5% of potato powder, 1.5% of yeast extract and 0.005% of magnesium sulfate into the pretreated molasses alcohol waste liquid, adjusting the pH value to 6.8 after mixing, sterilizing at 121 ℃ for 20min, inoculating 5% -8% of cyclic bacillus seed liquid into the mixed solution after the mixed solution is cooled to room temperature, controlling the fermentation temperature to be 30 ℃, controlling the dissolved oxygen to be 60% for batch fermentation, sterilizing for 33 hours after fermentation, removing filter residues after filtration, centrifuging the filtrate to obtain precipitate A and separating liquid B, and continuing secondary fermentation of the centrifuged separating liquid;
3) two-stage fermentation of fermentation waste liquid: adding 1.5% of peptone powder, 1.5% of bran powder and 0.005% of monopotassium phosphate into the centrifuged separation solution, adjusting the pH value of the mixture to 5.2-6.5, sterilizing at 121 ℃ for 20min, cooling the mixed solution to room temperature, inoculating 9% of lactobacillus casei-like seed solution into the mixed solution, controlling the fermentation temperature to be 33 ℃, controlling the dissolved oxygen to be 60%, performing batch fermentation, sterilizing for 26 hours, filtering, removing filter residues, centrifuging the filtrate to obtain a precipitate C and a separation solution D;
4) and (3) extracting polysaccharide: concentrating the separated liquid D to 1/7 of the original volume by a rotary evaporator, adding 0.2% alpha-amylase, carrying out enzymolysis for 25 min under the assistance of ultrasonic waves at the temperature of 30 ℃ and the pH value of 6.2, boiling to inactivate the enzyme after the enzymolysis is finished, centrifuging the enzymolysis liquid, adding 90% ethanol water solution into the centrifuged separated liquid to obtain a precipitate, and washing and drying the precipitate to obtain the wheat polysaccharide;
5) extraction of active peptide: mixing the precipitate A and the precipitate C, drying, pulverizing, sieving with 80 mesh sieve, adding 10 times of water, adjusting pH to 10.4 with sodium hydroxide, stirring at 60-75 deg.C for 1.5 hr, freezing, centrifuging for 5min, and adding oxalic acid to the supernatant to adjust pH to 4.5; freezing and centrifuging for 10min, removing supernatant, collecting precipitate, adding 6 times of distilled water, controlling solution temperature at 38 deg.C, adding 0.05% protease composition, performing enzymatic hydrolysis for 2.5 hr, heating to 80 deg.C, and inactivating enzyme to obtain solution containing active peptide.
The recovery rate of polysaccharide in the process is 40%, and the recovery rate of active peptide is 60%.
Example 3
A new process for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid comprises the following steps:
1) pretreatment of fermentation waste liquid: filtering the molasses alcohol waste liquid by using a coarse grating, filtering by using an ultrafiltration membrane with the diameter of 0.25 mu m, adding 0.1 percent of chitosan into the filtrate, strongly stirring, standing for 24 hours, carrying out ultrasonic treatment on the fermentation waste liquid for 5min, and filtering to obtain the pretreated molasses alcohol waste liquid;
2) primary fermentation of fermentation waste liquid: adding 0.5-2% of potato powder, 2% of yeast extract and 0.005% of magnesium sulfate into the pretreated molasses alcohol waste liquid, adjusting the pH value to 7.5 after mixing, sterilizing at 121 ℃ for 20min, inoculating 8% of cyclic bacillus seed liquid into the mixed solution after the mixed solution is cooled to room temperature, controlling the fermentation temperature to 32 ℃, controlling the dissolved oxygen to be 70% for batch fermentation, sterilizing for 35 hours after fermentation, removing filter residues after filtration, centrifuging the filtrate to obtain a precipitate A and a separating liquid B, and continuing secondary fermentation of the centrifuged separating liquid;
3) two-stage fermentation of fermentation waste liquid: adding 2% of peptone powder, 2% of bran powder and 0.005% of potassium dihydrogen phosphate into the centrifuged separation solution, adjusting the pH value of the mixture to 5.2-6.5, sterilizing at 121 ℃ for 20min, cooling the mixed solution to room temperature, inoculating 10% of lactobacillus casei-like seed solution into the mixed solution, controlling the fermentation temperature to be 34 ℃, controlling dissolved oxygen to be 70% for batch fermentation, sterilizing for 28 hours after fermentation, filtering, removing filter residues, and centrifuging the filtrate to obtain a precipitate C and a separation solution D;
4) and (3) extracting polysaccharide: concentrating the separated liquid D to 1/8 of the original volume by a rotary evaporator, adding 0.3% alpha-amylase, carrying out enzymolysis for 30 min under the assistance of ultrasonic waves at the temperature of 30 ℃ and at the pH value of 6.2, boiling to inactivate the enzyme after the enzymolysis is finished, centrifuging the enzymolysis liquid, adding 90% ethanol water solution into the centrifuged separated liquid to obtain a precipitate, and washing and drying the precipitate to obtain the wheat polysaccharide;
5) extraction of active peptide: mixing the precipitate A and the precipitate C, drying, pulverizing, sieving with 80 mesh sieve, adding 10 times of water, adjusting pH to 11.0 with sodium hydroxide, stirring in 75 deg.C water bath for 2 hr, freezing, centrifuging for 5min, and adding oxalic acid to the supernatant to adjust pH to 4.8; freezing and centrifuging for 10min, removing supernatant, collecting precipitate, adding 6 times of distilled water, controlling solution temperature at 40 deg.C, adding 0.05% protease composition, performing enzymatic hydrolysis for 3 hr, heating to 80 deg.C, and inactivating enzyme to obtain solution containing active peptide.
The recovery rate of polysaccharide is 35% and the recovery rate of active peptide is 53%
The above description is made in detail for the preferred embodiments of the present invention, but the above description is only for the preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention. All equivalent changes and modifications made within the scope of the present invention shall fall within the scope of the present invention.
Claims (3)
1. A method for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid is characterized by comprising the following steps:
1) pretreatment of fermentation waste liquid: filtering the molasses alcohol waste liquid by using a coarse grating, filtering by using an ultrafiltration membrane with the diameter of 0.25 mu m, adding 0.1 percent of chitosan into the filtrate, strongly stirring, standing for 20-24h, carrying out ultrasonic treatment on the fermentation waste liquid for 5min, and filtering to obtain the pretreated molasses alcohol waste liquid;
2) primary fermentation of fermentation waste liquid: adding 0.5-2% of potato powder, 1-2% of yeast extract and 0.005% of magnesium sulfate into the pretreated molasses alcohol waste liquid, adjusting the pH value to 6.2-7.5 after mixing, sterilizing for 20min at 121 ℃, cooling the mixed solution to room temperature, inoculating 5-8% of cyclic bacillus seed liquid into the mixed solution, controlling the fermentation temperature to be 28-32 ℃, controlling the dissolved oxygen to be 50-70% for batch fermentation, sterilizing for 30-35 hours after fermentation, filtering, removing filter residues, centrifuging the filtrate to obtain a precipitate A and a separating liquid B, and continuing secondary fermentation of the centrifuged separating liquid;
3) two-stage fermentation of fermentation waste liquid: adding 0.5-2% of peptone powder, 1-2% of bran powder and 0.005% of monopotassium phosphate into the centrifuged separation solution, adjusting the pH value of the mixture to 5.2-6.5, sterilizing at 121 ℃ for 20min, cooling the mixed solution to room temperature, inoculating 8-10% of lactobacillus casei-like seed solution into the mixed solution, controlling the fermentation temperature to be 32-34 ℃, controlling the dissolved oxygen to be 50-70% for batch fermentation, sterilizing for 24-28 hours, filtering, removing filter residues, and centrifuging the filtrate to obtain a precipitate C and a separation solution D;
4) and (3) extracting polysaccharide: concentrating the separation liquid D to 1/6-1/8 of the original volume through a rotary evaporator, adding 0.1-0.3% of alpha-amylase, performing enzymolysis for 20-30 min under the assistance of ultrasonic waves at the temperature of 30 ℃ and at the pH value of 6.2, boiling to inactivate the enzyme after the enzymolysis is finished, centrifuging the enzymolysis liquid, adding 90% of ethanol water solution into the centrifuged separation liquid to obtain precipitate, and washing and drying the precipitate to obtain the wheat polysaccharide;
5) extraction of active peptide: mixing the precipitate A and the precipitate C, drying, pulverizing, sieving with 80 mesh sieve, adding 10 times of water, adjusting pH to 9.5-11.0 with sodium hydroxide, stirring in a constant temperature water bath at 60-75 deg.C for 1-2 hr, freeze centrifuging for 5min, adding oxalic acid into the supernatant to adjust pH to 4.2-4.8; freezing and centrifuging for 10min, removing supernatant, collecting precipitate, adding 6 times of distilled water into the precipitate, controlling the temperature of the solution at 35-40 deg.C, adding 0.05% protease composition, performing enzymatic hydrolysis for 1.5-3 hr, heating to 80 deg.C, and inactivating enzyme to obtain solution containing active peptide;
the lactobacillus paracasei and the ring bacillus are in logarithmic growth phase when being accessed, and the colony number in the lactobacillus paracasei seed solution is not less than 2 multiplied by 109The number of bacterial colonies in the bacillus circulans seed liquid is not less than 5 multiplied by 109One per ml.
2. The method for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid as claimed in claim 1, wherein the seed culture medium of the bacillus circulans comprises: peptone 5.0g, beef extract 3.0g, NaCl5.0g, MnSO4·H2O5 mg, distilled water 1.0L, pH 7.0.
3. The method for co-producing wheat polysaccharide and active peptide by using alcohol fermentation waste liquid as claimed in claim 1, wherein the seed culture medium comprises casein peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, Tween 801.0 g, K2HPO42.0g，MgSO4.7H2O 0.2g，MnSO4.H2O0.05g, distilled water 1.0L, pH6.8.
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| CN1297994A (en) * | 2000-12-08 | 2001-06-06 | 曾文伟 | Production process of fungus powder and fungus polysaccharide for food and medicine |
| CN101709308A (en) * | 2009-10-22 | 2010-05-19 | 河南健古生物工程有限公司 | Method for producing fungus powder and fungus polysaccharide by using wheat alcohol waste lees solution |
| CN105580977A (en) * | 2014-10-24 | 2016-05-18 | 洛阳碧博生物科技有限公司 | Process for producing high protein feed from fermentation waste liquid |
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| CN1297994A (en) * | 2000-12-08 | 2001-06-06 | 曾文伟 | Production process of fungus powder and fungus polysaccharide for food and medicine |
| CN101709308A (en) * | 2009-10-22 | 2010-05-19 | 河南健古生物工程有限公司 | Method for producing fungus powder and fungus polysaccharide by using wheat alcohol waste lees solution |
| CN105580977A (en) * | 2014-10-24 | 2016-05-18 | 洛阳碧博生物科技有限公司 | Process for producing high protein feed from fermentation waste liquid |
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Denomination of invention: A new process for co producing wheat polysaccharides and active peptides using alcohol fermentation waste liquid Effective date of registration: 20231124 Granted publication date: 20200121 Pledgee: Binzhou Bincheng Branch of China Construction Bank Co.,Ltd. Pledgor: BINZHOU ZHONGYU FOOD Co.,Ltd. Registration number: Y2023370000133 |
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