CN111011707A - Preparation method of flavor detoxified corn steep liquor - Google Patents
Preparation method of flavor detoxified corn steep liquor Download PDFInfo
- Publication number
- CN111011707A CN111011707A CN201911194779.4A CN201911194779A CN111011707A CN 111011707 A CN111011707 A CN 111011707A CN 201911194779 A CN201911194779 A CN 201911194779A CN 111011707 A CN111011707 A CN 111011707A
- Authority
- CN
- China
- Prior art keywords
- steep liquor
- corn steep
- zen
- don
- degrading
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 240000008042 Zea mays Species 0.000 title claims abstract description 207
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 207
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 207
- 235000005822 corn Nutrition 0.000 title claims abstract description 207
- 238000002360 preparation method Methods 0.000 title claims abstract description 38
- 239000000796 flavoring agent Substances 0.000 title claims abstract description 27
- 235000019634 flavors Nutrition 0.000 title claims abstract description 26
- 230000000593 degrading effect Effects 0.000 claims abstract description 118
- 238000000855 fermentation Methods 0.000 claims abstract description 96
- 230000004151 fermentation Effects 0.000 claims abstract description 96
- 241000894006 Bacteria Species 0.000 claims abstract description 70
- 108090000790 Enzymes Proteins 0.000 claims abstract description 63
- 102000004190 Enzymes Human genes 0.000 claims abstract description 63
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 52
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 38
- 239000003053 toxin Substances 0.000 claims abstract description 36
- 231100000765 toxin Toxicity 0.000 claims abstract description 36
- 238000006731 degradation reaction Methods 0.000 claims abstract description 35
- 239000004310 lactic acid Substances 0.000 claims abstract description 27
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 26
- 241000186660 Lactobacillus Species 0.000 claims abstract description 11
- 150000001413 amino acids Chemical class 0.000 claims abstract description 11
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 11
- 238000010170 biological method Methods 0.000 claims abstract description 5
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 35
- 238000000034 method Methods 0.000 claims description 35
- 230000015556 catabolic process Effects 0.000 claims description 34
- 239000007788 liquid Substances 0.000 claims description 34
- 244000063299 Bacillus subtilis Species 0.000 claims description 30
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 30
- 239000001963 growth medium Substances 0.000 claims description 29
- 241000228245 Aspergillus niger Species 0.000 claims description 23
- 238000004321 preservation Methods 0.000 claims description 22
- 238000004519 manufacturing process Methods 0.000 claims description 17
- 238000005507 spraying Methods 0.000 claims description 17
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 12
- 230000004913 activation Effects 0.000 claims description 11
- 238000009629 microbiological culture Methods 0.000 claims description 11
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 9
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 9
- 235000001014 amino acid Nutrition 0.000 claims description 9
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 9
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 4
- 235000013922 glutamic acid Nutrition 0.000 claims description 4
- 239000004220 glutamic acid Substances 0.000 claims description 4
- 239000004471 Glycine Substances 0.000 claims description 3
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 3
- 239000013604 expression vector Substances 0.000 claims description 2
- 235000012054 meals Nutrition 0.000 claims description 2
- 239000006041 probiotic Substances 0.000 claims description 2
- 230000000529 probiotic effect Effects 0.000 claims description 2
- 235000018291 probiotics Nutrition 0.000 claims description 2
- 238000003259 recombinant expression Methods 0.000 claims description 2
- 235000015099 wheat brans Nutrition 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 19
- 230000000050 nutritive effect Effects 0.000 abstract description 2
- MBMQEIFVQACCCH-QBODLPLBSA-N zearalenone Chemical compound O=C1O[C@@H](C)CCCC(=O)CCC\C=C\C2=CC(O)=CC(O)=C21 MBMQEIFVQACCCH-QBODLPLBSA-N 0.000 description 89
- MBMQEIFVQACCCH-UHFFFAOYSA-N trans-Zearalenon Natural products O=C1OC(C)CCCC(=O)CCCC=CC2=CC(O)=CC(O)=C21 MBMQEIFVQACCCH-UHFFFAOYSA-N 0.000 description 86
- LINOMUASTDIRTM-QGRHZQQGSA-N deoxynivalenol Chemical compound C([C@@]12[C@@]3(C[C@@H](O)[C@H]1O[C@@H]1C=C(C([C@@H](O)[C@@]13CO)=O)C)C)O2 LINOMUASTDIRTM-QGRHZQQGSA-N 0.000 description 75
- LINOMUASTDIRTM-UHFFFAOYSA-N vomitoxin hydrate Natural products OCC12C(O)C(=O)C(C)=CC1OC1C(O)CC2(C)C11CO1 LINOMUASTDIRTM-UHFFFAOYSA-N 0.000 description 73
- 229930002954 deoxynivalenol Natural products 0.000 description 71
- 108700012359 toxins Proteins 0.000 description 32
- 230000001580 bacterial effect Effects 0.000 description 24
- 239000002068 microbial inoculum Substances 0.000 description 24
- 230000003213 activating effect Effects 0.000 description 22
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 20
- 239000000126 substance Substances 0.000 description 20
- 231100000678 Mycotoxin Toxicity 0.000 description 19
- 239000002636 mycotoxin Substances 0.000 description 19
- 239000000463 material Substances 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 15
- 229910052760 oxygen Inorganic materials 0.000 description 15
- 239000001301 oxygen Substances 0.000 description 15
- 238000012216 screening Methods 0.000 description 15
- 239000000047 product Substances 0.000 description 13
- 238000012163 sequencing technique Methods 0.000 description 13
- 239000000243 solution Substances 0.000 description 13
- 239000006228 supernatant Substances 0.000 description 13
- 239000001888 Peptone Substances 0.000 description 12
- 108010080698 Peptones Proteins 0.000 description 12
- 235000019319 peptone Nutrition 0.000 description 12
- 230000001954 sterilising effect Effects 0.000 description 12
- 238000012258 culturing Methods 0.000 description 11
- 239000000843 powder Substances 0.000 description 11
- 230000008569 process Effects 0.000 description 11
- 239000011378 shotcrete Substances 0.000 description 11
- 239000002002 slurry Substances 0.000 description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 238000001035 drying Methods 0.000 description 10
- 239000002609 medium Substances 0.000 description 10
- 238000002156 mixing Methods 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 238000003556 assay Methods 0.000 description 9
- 239000010903 husk Substances 0.000 description 9
- 239000012528 membrane Substances 0.000 description 9
- 239000006188 syrup Substances 0.000 description 9
- 235000020357 syrup Nutrition 0.000 description 9
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 8
- 229940024606 amino acid Drugs 0.000 description 8
- 238000004128 high performance liquid chromatography Methods 0.000 description 8
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 8
- 239000010865 sewage Substances 0.000 description 8
- 239000007836 KH2PO4 Substances 0.000 description 7
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 7
- 229960003767 alanine Drugs 0.000 description 7
- 229910052564 epsomite Inorganic materials 0.000 description 7
- 238000001704 evaporation Methods 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 235000013312 flour Nutrition 0.000 description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N D-alpha-Ala Natural products CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 6
- QNAYBMKLOCPYGJ-UWTATZPHSA-N L-Alanine Natural products C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 6
- 230000009286 beneficial effect Effects 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 230000000877 morphologic effect Effects 0.000 description 6
- 229940073490 sodium glutamate Drugs 0.000 description 6
- 238000011144 upstream manufacturing Methods 0.000 description 6
- 238000012512 characterization method Methods 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 239000003085 diluting agent Substances 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 235000013379 molasses Nutrition 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 239000010802 sludge Substances 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 238000003860 storage Methods 0.000 description 5
- 230000004544 DNA amplification Effects 0.000 description 4
- 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 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 241000235342 Saccharomycetes Species 0.000 description 4
- 238000000137 annealing Methods 0.000 description 4
- 239000000919 ceramic Substances 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 238000004925 denaturation Methods 0.000 description 4
- 230000036425 denaturation Effects 0.000 description 4
- 229910000388 diammonium phosphate Inorganic materials 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 239000003337 fertilizer Substances 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 230000007062 hydrolysis Effects 0.000 description 4
- 238000006460 hydrolysis reaction Methods 0.000 description 4
- 235000016709 nutrition Nutrition 0.000 description 4
- 235000019629 palatability Nutrition 0.000 description 4
- 238000012257 pre-denaturation Methods 0.000 description 4
- 238000000108 ultra-filtration Methods 0.000 description 4
- 238000009423 ventilation Methods 0.000 description 4
- NOIIUHRQUVNIDD-UHFFFAOYSA-N 3-[[oxo(pyridin-4-yl)methyl]hydrazo]-N-(phenylmethyl)propanamide Chemical compound C=1C=CC=CC=1CNC(=O)CCNNC(=O)C1=CC=NC=C1 NOIIUHRQUVNIDD-UHFFFAOYSA-N 0.000 description 3
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 3
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- 241000223218 Fusarium Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000003794 Gram staining Methods 0.000 description 3
- 239000013543 active substance Substances 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000007664 blowing Methods 0.000 description 3
- 229940041514 candida albicans extract Drugs 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 239000003205 fragrance Substances 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 238000001694 spray drying Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000012138 yeast extract Substances 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108010068370 Glutens Proteins 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000005913 Maltodextrin Substances 0.000 description 2
- 229920002774 Maltodextrin Polymers 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- RAHZWNYVWXNFOC-UHFFFAOYSA-N Sulphur dioxide Chemical compound O=S=O RAHZWNYVWXNFOC-UHFFFAOYSA-N 0.000 description 2
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940011871 estrogen Drugs 0.000 description 2
- 239000000262 estrogen Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- -1 fusarium graminearum Chemical compound 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 235000021312 gluten Nutrition 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 229910017053 inorganic salt Inorganic materials 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 229940035034 maltodextrin Drugs 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 229930000044 secondary metabolite Natural products 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 239000004065 semiconductor Substances 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- CWERGRDVMFNCDR-UHFFFAOYSA-N thioglycolic acid Chemical compound OC(=O)CS CWERGRDVMFNCDR-UHFFFAOYSA-N 0.000 description 2
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 description 1
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 1
- 108050001049 Extracellular proteins Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000223195 Fusarium graminearum Species 0.000 description 1
- 241000221779 Fusarium sambucinum Species 0.000 description 1
- 108091023242 Internal transcribed spacer Proteins 0.000 description 1
- 241000235058 Komagataella pastoris Species 0.000 description 1
- 241001506991 Komagataella phaffii GS115 Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 206010000210 abortion Diseases 0.000 description 1
- 231100000176 abortion Toxicity 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000002856 computational phylogenetic analysis Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000001054 cortical effect Effects 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 238000005485 electric heating Methods 0.000 description 1
- 231100000049 endocrine disruptor Toxicity 0.000 description 1
- 239000000598 endocrine disruptor Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229930182833 estradiol Natural products 0.000 description 1
- 229960005309 estradiol Drugs 0.000 description 1
- RESSOZOGQXKCKT-UHFFFAOYSA-N ethene;propane-1,2-diol Chemical compound C=C.CC(O)CO RESSOZOGQXKCKT-UHFFFAOYSA-N 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- 231100000206 health hazard Toxicity 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 235000021395 porridge Nutrition 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000013630 prepared media Substances 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000004455 soybean meal Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229960003604 testosterone Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000007738 vacuum evaporation Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
- 239000007222 ypd medium Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
-
- 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
-
- 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
- C12N1/16—Yeasts; Culture media therefor
-
- 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
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Virology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Botany (AREA)
- Medicinal Chemistry (AREA)
- Nutrition Science (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Animal Husbandry (AREA)
- Sustainable Development (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The embodiment of the invention discloses a preparation method of flavor detoxified corn steep liquor, which is characterized in that DON degrading bacteria or degrading enzyme is added into the corn steep liquor, and DON toxin is degraded by a biological method; adding ZEN degrading bacteria or degrading enzyme into the corn steep liquor, and degrading ZEN toxin by a biological method; adding lactobacillus and yeast into the corn steep liquor for fermentation, improving the content of amino acid and lactic acid in the corn steep liquor, and improving the flavor of the corn steep liquor to obtain the flavor-detoxified corn steep liquor. The ZEN content in the detoxified corn steep liquor is 0-100 mug/kg, the DON content is 0-500 mug/kg, and the detoxified effect of the corn steep liquor is good; simultaneously, yeast and lactic acid bacteria are added in the ZEN and DON degradation process, and the content of various amino acids is increased and a certain amount of lactic acid is generated in the fermentation process of the corn steep liquor, so that the corn steep liquor is more palatable, the nutritive value of the corn steep liquor is improved, and the flavor of the corn steep liquor is improved.
Description
Technical Field
The embodiment of the invention relates to the technical field of biology, and particularly relates to a preparation method of flavor-detoxified corn steep liquor.
Background
Deoxynivalenol (DON) is also called vomitoxin, is a toxic secondary metabolite generated by some fusarium, is usually produced by mildew in the links of grain growth, harvesting, storage, processing, transportation, sale and the like, and can cause breeding performance disorders such as abolishment of appetite of livestock and poultry, growth obstruction, metabolic disturbance, immune disorder, female animal sterility, pregnant animal abortion, fetal abnormality and the like, and cause serious damage to the animal husbandry.
Zearalenone (ZEN) also called F-2 toxin is a secondary metabolite of non-steroid estrogen produced by a plurality of fusarium such as fusarium graminearum, fusarium sojae, fusarium roseum and the like, ZEN shows strong estrogen effect on livestock, particularly female pigs, and is very easy to cause serious reproductive disorders; the zearalenone also has an interference effect on the secretion system in a human body, and researches such as Fizzell and the like show that when the zearalenone concentration is 100 mu mol/L, the levels of estradiol, testosterone and cortical hormone are remarkably reduced, and ZEN and metabolites thereof change the production of hormone by influencing receptor signals and are potential endocrine disruptors. In addition, because the corn steep liquor contains a large amount of sulfurous acid, pungent smell can be generated, the palatability is influenced, and the addition amount of the corn steep liquor in daily ration is reduced.
Two toxins, namely DON and ZEN, generally exist in corn steep liquor prepared by the prior art, but the prior art does not have a method for effectively removing the two toxins in the corn steep liquor, so that health hazards are brought to people and animals.
Disclosure of Invention
Therefore, the embodiment of the invention provides a preparation method of a corn steep liquor with a detoxified flavor, which aims to solve the problem that DON and ZEN toxins in the corn steep liquor cannot be effectively detoxified in the prior art and realize the improvement of the flavor of the corn steep liquor by cooperating with lactic acid bacteria and yeast.
In order to achieve the above object, the embodiments of the present invention provide the following technical solutions:
a method for preparing flavor detoxified corn steep liquor comprises adding DON degrading bacteria or degrading enzyme into corn steep liquor, and biologically degrading DON toxin;
adding ZEN degrading bacteria or degrading enzyme into the corn steep liquor, and degrading ZEN toxin by a biological method;
adding lactobacillus and yeast into the corn steep liquor for fermentation, improving the content of amino acid and lactic acid in the corn steep liquor, and improving the flavor of the corn steep liquor to obtain the flavor-detoxified corn steep liquor.
Preferably, the DON degrading bacteria are Bacillus subtilis with the Latin name of Bacillus subtilis, and the Bacillus subtilis is preserved in China general microbiological culture Collection center in 2018, 08 and 13 months with the preservation number of CGMCC NO.16254 and the preservation address of: the institute of microbiology, national academy of sciences No. 3, Xilu No.1, Beijing Korean district, China;
the ZEN degrading bacteria are Aspergillus niger, the Latin name of the ZEN degrading bacteria is Aspergillus niger, the ZEN degrading bacteria are preserved in China general microbiological culture Collection center (CGMCC) in 11-23 th of 2017, the preservation number is CGMCC NO.14639, and the preservation address is as follows: the institute of microbiology, China academy of sciences, No. 3, Xilu No.1, Beijing Korean district, Beichen, China.
Preferably, the Lactobacillus is Lactobacillus plantarum, the latin name of which is Lactobacillus plantarum, the Lactobacillus is deposited in the common microorganism center of the China Committee for culture Collection of microorganisms in 2018, 08 and 14 months, the deposit number is CGMCC NO.16256, and the deposit address is: the institute of microbiology, national academy of sciences No. 3, Xilu No.1, Beijing Korean district, China;
the yeast is Saccharomyces cerevisiae, the Latin name of the Saccharomyces cerevisiae is Saccharomyces cerevisiae, the strain is preserved in China general microbiological culture Collection center in 13.09.2018, the preservation number is CGMCC NO.16469, and the preservation address is as follows: the institute of microbiology, China academy of sciences, No. 3, Xilu No.1, Beijing Korean district, Beichen, China.
Preferably, the step of adding ZEN degrading bacteria or ZEN degrading enzyme into the corn steep liquor and the step of adding DON degrading bacteria or DON enzyme for degradation specifically comprises the following steps:
adding any one of ZEN degrading bacteria and DON degrading bacteria, ZEN degrading bacteria and DON degrading enzyme, ZEN degrading enzyme and DON degrading bacteria, ZEN degrading enzyme and DON degrading enzyme, ZEN degrading bacteria and DON degrading enzyme into the corn steep liquor, wherein ZEN degrading and DON degrading can be carried out step by step or synchronously.
Preferably, the ZEN content in the flavor detoxified corn steep liquor is lower than 100 mug/kg, the DON content is lower than 500 mug/kg, and the probiotic content is more than or equal to 109CFU/ml, increased lactic acid, glutamic acid and glycine content.
Preferably, the ZEN and DON degrading enzymes are in a solid enzyme preparation or a liquid enzyme preparation;
the ZEN and DON degrading bacteria are solid bacteria preparation or liquid activated seed liquid.
Preferably, the ZEN and DON degrading bacteria are activated by respectively inoculating the ZEN and DON degrading bacteria into a fermentation tank containing an activation culture medium and activating for 2-3 hours;
wherein the rotating speed of the fermentation tank is 100-.
Preferably, the degrading enzyme is prepared by fermenting degrading bacteria or is introduced into a host cell to obtain a recombinant expression vector with a ZEN degrading enzyme coding gene, and then the ZEN and DON degrading enzyme is obtained by the expression of the host cell.
The embodiment of the invention also provides a preparation method of the flavor feed product, and the flavor detoxified corn steep liquor prepared by the method is sprayed on the feed product.
Preferably, the feed product comprises corn bran, wheat bran or corn germ meal.
The embodiment of the invention has the following advantages:
the detoxified corn steep liquor utilizes ZEN degrading bacteria and ZEN degrading enzyme to carry out enzymolysis treatment on ZEN and DON mycotoxin existing in the corn steep liquor, and simultaneously, lactobacillus and saccharomycetes are added into the corn steep liquor for fermentation, so that DON and ZEN detoxified corn steep liquor with flavors can be obtained, and the detoxication effect on the corn steep liquor is good; meanwhile, in the fermentation process of the lactic acid bacteria and the saccharomycetes, the amino acid content is obviously increased, and a certain amount of lactic acid is generated, so that the corn steep liquor is more palatable, the nutritive value of the corn steep liquor is improved, the flavor of the corn steep liquor is improved, the corn steep liquor treated by ZEN degrading bacteria and/or ZEN degrading enzymes and DON degrading enzymes can be further prepared after the corn steep liquor is subjected to slurry spraying treatment on the corn bran, and the slurry sprayed corn bran has a good application prospect on preparation of flavor feed.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
FIG. 1 is a photograph showing the characteristics of TD-2 colonies of DON-degrading bacteria according to an embodiment of the present invention;
FIG. 2 is a graph of a tree-evolved analysis of DON-degrading bacteria TD-2 according to an embodiment of the present invention;
FIG. 3 is a photograph showing the colony characteristics of degraded ZEN Aspergillus niger strain FS-7 provided by the embodiment of the present invention;
FIG. 4 is an analysis diagram of an FS-7 evolutionary tree of a strain Aspergillus niger degrading strain provided by the embodiment of the invention;
FIG. 5 is a photograph of a colony of yeast strain TM-3 provided in an embodiment of the present invention;
FIG. 6 is a graph of a phylogenetic tree analysis of the yeast strain TM-3 provided in an embodiment of the present invention;
FIG. 7 is a photographic image of colonies of lactic acid bacteria strain RS-5 provided in the examples of the present invention;
FIG. 8 is a chart of an RS-5 strain evolutionary tree analysis of the lactic acid bacteria strain provided by the embodiment of the present invention.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the examples of the present invention, all percentages, parts and ratios are based on the total weight of the composition of the present invention, unless otherwise specified. All weights as they pertain to listed ingredients are based on the active level and, therefore, do not include solvents or by-products found in commercially available materials, unless otherwise specified.
In the embodiment of the invention, the corn steep liquor is prepared by soaking corn particles in a sulfur dioxide solution in a countercurrent way, finally pumping out a solution with soaking water containing 6-7% of dry substances, and then concentrating the solution to a concentrated solution with a certain amount of dry substances by adopting three-effect vacuum evaporation, wherein the concentrated solution is called the corn steep liquor.
In the present embodiment, the term "dry substance" refers to the dry substance remaining after the corn steep liquor is dried.
In the embodiment of the invention, the spraying is to mix the corn steep liquor and the corn bran, dry the corn steep liquor and the corn bran by using the tube bundle, and then crush the corn steep liquor and the corn bran to prepare the spraying corn bran.
In the embodiment of the invention, the ZEN degrading enzyme can be the ZEN degrading enzyme prepared in the embodiment of the invention, and can also be the ZEN degrading enzyme, and the ZEN degrading enzyme can be in a liquid state or a solid powder state.
In the embodiment of the invention, the bacillus subtilis refers to a strain with the capacity of degrading DON toxin, and particularly can secrete extracellular protein capable of effectively degrading vomitoxin, and has a high degradation rate on the DON toxin.
In the embodiment of the invention, the used equipment comprises 1 seed activation fermentation tank with the specification of 20L; 1 fermentation tank with the specification of 1 t; 1 evaporator with the specification of 1 t; 1 tube bundle dryer; 1, a plurality of fermentation bags of a crusher with the specification of 50 kg; 1 feed mixer.
Example 1 screening of Bacillus subtilis for DON degradation
1 materials and reagents: the sample for screening the bacillus subtilis comes from a sewage treatment pool, sludge, a wheat gluten production workshop, open soil, a fertilizer production workshop, river sludge, a corn processing workshop, sewage of a sewage treatment plant and a lysine production workshop.
1.1 reagents and media: chromatographic grade methanol, acetonitrile: thermal corporation; and (3) preparing a DON standard substance: sigma corporation. Inorganic salt medium (MSM): (NH)4)2SO40.5g，MgSO4·7H2O 0.2g，CaCl20.05g，Na2HPO42.44g，KH2PO41.52g, adding water to 1L, adjusting pH to 6.8, sterilizing at 121 deg.C for 20min, and adding 1.5% agar into solid culture medium. LB culture medium: 10g of peptone, 10g of NaCl, 5g of yeast extract, dissolved in 1L of water, and sterilized at 121 ℃ for 20 min.
1.2 main instruments and equipment: high performance liquid chromatography (e2695), ultraviolet detector (2998): water corporation, usa; a PCR instrument: BIO-RAD, USA; gel imager: BIO-RAD, USA;
2. the experimental method comprises the following steps: the DON content determination method refers to national standard GB5009.111-2016, and the high performance liquid chromatography conditions are as follows, and the chromatographic column comprises: c18(259 mm. times.4.6 mm, 5 μm, xbridge); mobile phase: water/methanol 80/20 (V/V); the flow rate is 0.8 mL/min; the column temperature is 35 ℃; the sample volume is 50 mu L; an ultraviolet detector: the detection wavelength was 218 nm.
2.2 separation and purification of degrading bacteria: enriching and primary screening, namely putting 5g of sample into 50mL of MSM culture medium, culturing for 10 days at 30 ℃ and 220r/min with the DON concentration of 20mg/L, inoculating for 3 times with 10% of inoculum size, detecting DON residue by high performance liquid chromatography, and selecting the sample with the DON degradation effect to carry out the next experiment. And (3) properly diluting and coating a sample with degradation effect obtained by primary screening on an MSM (metal-organic semiconductor) plate, selecting single colonies with different forms, inoculating the single colonies to an EP (EP) tube containing 10 mu g of DON, culturing for 6d at 30 ℃ and 220r/min, detecting DON residues, and selecting a strain with degradation effect to carry out next experiment.
2.3 identification of degrading bacteria: the obtained strain is streaked on an MSM plate, cultured at 30 ℃ for 24-48h, and observed for colony morphology, size, edge, surface bulge shape, transparency and the like. Gram staining of the strain was performed.
2.4 molecular identification: identifying the obtained degrading bacteria by 16SrDNA or 18SrDNA, taking a bacterial liquid or a single colony as a template, and reacting: template 1 uL, Taqmix 12.5 uL, 16S upstream and downstream primers 1 uL, ddH2O10.5. mu.L. Reaction procedure: pre-denaturation at 98 deg.C for 10min, denaturation at 94 deg.C for 50s, annealing at 55 deg.C for 45s, extension at 72 deg.C for 1.5min, 30 cycles, extension at 72 deg.C for 10min, and storage at 4 deg.C. And detecting the PCR product through nucleic acid electrophoresis, sending the PCR product to Invitrogen company for sequencing, and comparing the sequence obtained by sequencing on NCBI. Wherein, the bacterial universal primer SEQ ID NO: 1, 16SF (5'-attgaacgctggcggcaggcct-3'); SEQ ID NO: 2, 16SR (5'-taacaaggtaaccgtagggg a-3'). The general primers of the fungus are as follows: SEQ ID NO: 3, 18SF (5'-gtagtcatatgcttgtctc-3'); SEQ ID NO: 4, 18SR (5'-attccccgttacccgttg-3').
And 3, separating and purifying the degrading bacteria, and screening 9 factory samples according to the method to obtain 1 bacterial strain with the degrading effect on DON, wherein the bacterial strain is named as TD-2.
3.1 adding 500 mu L of the supernatant of the strain fermentation liquor and the crushed supernatant of the fermentation liquor into an EP tube containing 10 mu g of DON, reacting for 12h at 37 ℃ and 220r/min, measuring the DON residual quantity and calculating the degradation rate by using the crushed supernatant of the inactivated strain fermentation liquor as a reference through high performance liquid chromatography, wherein the results are shown in Table 1, and the degradation rate of the active substances of the DON degrading bacteria is shown.
TABLE 1
3.2 morphological characterization of the TD-2DON Strain, as shown in FIG. 1, the strain TD-2 is characterized by: the colony is flat, diffused, irregular in edge, rough and opaque in surface, light yellow and gram-positive.
3.3 molecular characterization of degrading strains: the bacterial liquid is used as a template, 16S universal primers are used for gene amplification, a PCR product is sequenced, and the sequencing result is compared in a GeneBank database, so that the homology between the bacterial strain TD-2 and the Bacillus amyloliquefaciens strain md1-51 is up to 99 percent. The result of the phylogenetic tree constructed by MEGA-5.0 software for TD-2 strain by N-J method is shown in FIG. 2. According to the similarity of the strain 16SrDNA, phylogenetic tree and colony morphology, the TD-2 strain is preliminarily identified as Bacillus subtilis strain. And finally, adding glycerol into the obtained bacterial strain capable of degrading the deoxynivalenol toxin, and storing in a refrigerator at the temperature of 70 ℃ below zero. The Latin of the Bacillus subtilis is named as Bacillus subtilis, and the Bacillus subtilis is preserved in China general microbiological culture Collection center (CGMCC) at 13.08.2018 with the preservation number of CGMCC NO.16254 and the preservation address of China institute of microbiology, China institute of academy of sciences, No. 3, West Lu 1, North Cheng of the Beijing Korean district, China.
3.4 fermenting and culturing the bacillus subtilis for 2-3h, adding 30% of argil filler into the fermentation broth to obtain a bacillus subtilis fermentation broth mixture, and then carrying out spray drying on the bacillus subtilis fermentation broth mixture to obtain the bacillus subtilis microbial inoculum for degrading DON.
Example 2 screening and identification of ZEN degrading Aspergillus niger
1, screening a sample for degrading ZEN Aspergillus niger from a sewage treatment pool, sludge, a wheat gluten production workshop, open land soil, a fertilizer production workshop, river sludge, a corn processing workshop, sewage of a sewage treatment plant, a lysine production workshop, sewage of an aerobic pool of the sewage treatment plant, fertilizer of a fertilizer plant, landscape river sludge, a glutamic acid production workshop and open land soil.
1.1 reagents and cultureAnd (3) nutrient medium: chromatographic grade methanol, acetonitrile: thermal corporation; ZEN standard substance: sigma corporation. Inorganic salt medium (MSM): (NH)4)2SO40.5g，MgSO4·7H2O 0.2g，CaCl20.05g，Na2HPO4，2.44g，KH2PO41.52g, adding water to 1L, adjusting pH to 6.8, sterilizing at 121 deg.C for 20min, and adding 1.5% agar into solid culture medium.
PDA culture medium: adding 20g of glucose into water to reach a constant volume of 200ml, and sterilizing at 121 ℃ for 20 min. Dicing 200g potato, adding water, boiling, filtering with eight layers of gauze, adding water to 800ml, sterilizing at 115 deg.C for 20min, and adding sterilized glucose solution.
1.2 main instruments and equipment: high performance liquid chromatography (e2695), fluorescence detector (2475): water corporation, usa; a PCR instrument: BIO-RAD, USA; gel imager: BIO-RAD, USA; agilent 1200-6510Q-TofLC/MS liquid chromatography-mass spectrometer: agilent, USA; a pulverizer: tianjin Tester instruments, Inc.; a pneumatic control operation frame: beijing Huaan Maike Biotechnology Ltd.
1.3 Experimental methods: the ZEN determination method refers to a national standard method GB5009.209-2016, and the high performance liquid chromatography conditions are as follows, wherein the method comprises the following steps: c18(259 mm. times.4.6 mm, 5 μm, xbridge); mobile phase: water/acetonitrile/methanol-46/46/8 (V/V); the flow rate is 1.0 mL/min; the column temperature is 25 ℃; the sample volume is 10 mu L; a fluorescence detector: the excitation wavelength (Ex) is 274nm and the emission wavelength (EM) is 440 nm. The ZEN degradation rate is ZEN content in the degraded medium/ZEN content in the original medium multiplied by 100%.
1.3.1 enrichment and Primary screening: taking 5g of sample to 50mL of MSM culture medium, culturing the sample at 30 ℃ and 220r/min for 10d with the ZEN concentration of 20mg/L, transferring for 3 times with the inoculation amount of 10%, detecting the ZEN residue by high performance liquid chromatography, and selecting the sample with the ZEN degradation effect to carry out the next experiment. Re-screening: and (3) coating a sample with degradation effect obtained by primary screening on an MSM (Metal-oxide-semiconductor) plate through proper dilution, selecting single colonies with different forms, inoculating the single colonies into an EP (ethylene propylene glycol) tube containing 10 mu g of ZEN, culturing for 6d under the conditions of 30 ℃ and 220r/min, detecting ZEN residue, and selecting a strain with degradation effect to carry out next experiment.
1.3.2 morphological characterization of degrading bacterial strains: the obtained strain is streaked on an MSM plate, cultured at 30 ℃ for 24-48h, and observed for colony morphology, size, edge, surface bulge shape, transparency and the like. Gram staining of the strain was performed.
And (3) molecular identification: identifying the obtained degrading bacteria by 16SrDNA or 18SrDNA, taking a bacterial liquid or a single colony as a template, and reacting: template 1 uL, Taqmix 12.5 uL, 16S upstream and downstream primers 1 uL, ddH2O10.5. mu.L. Reaction procedure: pre-denaturation at 98 deg.C for 10min, denaturation at 94 deg.C for 50s, annealing at 55 deg.C for 45s, extension at 72 deg.C for 1.5min, 30 cycles, extension at 72 deg.C for 10min, and storage at 4 deg.C. And detecting the PCR product through nucleic acid electrophoresis, sending the PCR product to Invitrogen company for sequencing, and comparing the sequence obtained by sequencing on NCBI. Wherein the sequence of the bacterial universal primer is shown as SEQ ID NO: 1. SEQ ID NO: 2 is shown in the specification; the fungal universal primer sequence is shown as SEQ ID NO: 3, SEQ ID NO: 4, respectively.
2, separating and purifying degrading bacteria, and screening 18 factory samples according to the method of 1.3.1 to obtain 1 strain FS-7 with the degrading effect on ZEN.
2.1 taking 500 mu L of the strain fermentation liquor supernatant and the fermentation liquor crushed supernatant, adding the strain fermentation liquor supernatant and the fermentation liquor crushed supernatant into an EP tube containing 10 mu g of ZEN, reacting for 12h at 30 ℃ and 220r/min, taking the inactivated strain fermentation liquor crushed supernatant as a reference, measuring the ZEN residual quantity by high performance liquid chromatography and calculating the degradation rate, wherein the results are shown in a table 2, and the secretion characteristic of the strain active substances is researched. As can be seen from Table 2, the degradation rate of the broken supernatant of the FS-7 fermentation broth is more than 70%, while the degradation rate of the fermentation supernatant is only 24%, which indicates that the active substances are mainly distributed in cells.
TABLE 2
2.2 morphological identification of FS-7 degrading bacteria, wherein hyphae are developed, black brown and thick velvet and are distributed on the whole dish, black spores are attached to the surface, and colonies are not easy to pick up, as shown in figure 3.
2.2.1 molecular characterization: and (3) performing gene amplification by using the bacterial liquid as a template and using 18S universal primers, sequencing the PCR product, and comparing the sequencing result in a GeneBank database, wherein the homology between the FS-7 strain and Aspergillus niger is up to 99 percent. The result of constructing a phylogenetic tree of the FS-7 strain by the N-J method using MEGA-5.0 software is shown in FIG. 4. According to the similarity of the strain 18SrDNA, phylogenetic tree and colony morphology, the FS-7 strain is preliminarily identified as Aspergillus niger, the Latin article name of the Aspergillus niger is Aspergillus niger, the strain is preserved in China general microbiological culture Collection center at 11 months and 23 days in 2017, the preservation number is CGMCC NO.14639, and the preservation address is China academy of sciences microbial research institute No. 3, North Cheng West Lu No.1 institute of China, North Jing rising district, China.
Example 3 screening of yeasts for flavor-enhancing substances
1, materials and methods: the sample sources were food mills, flour mills, breweries and wineries.
1.1 reagents and media: amino acid standards, Wako corporation, japan; hydrochloric acid (guaranteed purity), tianjin reagent; thioglycolic acid, Sigma, usa; mobile phase (ninhydrin solution and buffer) for amino acid analyzer, Wako corporation, japan. The enrichment culture medium comprises 20g/L of glucose, 20g/L of peptone, 10g/L of yeast extract and 100mg/L of penicillin. YPD medium comprises peptone 20g/L, yeast extract 10g/L, and glucose 20 g/L.
1.2 main instruments and equipment: model L-8900 amino acid analyzer, Hitachi, Japan; electric heating constant temperature drying oven, shanghai Senxin; concentrator, tokyo physical & chemical company; nitrogen blowing apparatus, Organomation, USA
1.3 Experimental methods: separating and purifying the yeast, and enriching: 1g of the sample was placed in 50mL of enrichment medium and cultured with shaking at 25 ℃ for 16 hours. Separation and purification: diluting the enrichment solution to 10-3, 10-4 and 10-5 in a gradient manner, taking 0.1mL of dilution solution in a separation culture medium, coating a flat plate, culturing at 30 ℃, observing the growth condition of bacterial colonies, and after the bacterial colonies grow out, carrying out streak purification on the bacterial colonies of which the thalli are saccharomycetes.
1.3.1 sample treatment: evaporating 5ml of fermentation liquor to dryness, dissolving the fermentation liquor by using 1ml of ultrapure water, adding 2.0ml of concentrated hydrochloric acid, uniformly mixing 2-3 drops of phenol, vacuumizing, blowing nitrogen by using a nitrogen blowing instrument, carrying out glass hot melting sealing on a glass hydrolysis tube, placing the sealed hydrolysis tube in a 110 ℃ oven, hydrolyzing for 24 hours, taking out the hydrolysis tube, cooling to room temperature, transferring the hydrolyzed sample into a 25ml volumetric flask, repeatedly cleaning the hydrolysis tube by using pure water, and fixing the volume of the volumetric flask to a scale. And (3) putting the sample after volume fixing into a sample injection small bottle to be detected after passing through a 0.22 mu m microporous filter membrane.
1.3.2 morphological identification, observing colony characteristics: size, texture, color, whether the surface is smooth, whether the edges are neat, cell shape, whether the arrangement is regular, division pattern, etc.
Molecular biology identification, namely identifying 5.8SrDNA of the obtained saccharomycetes, taking a bacterial liquid or a single colony as a template, and reacting: template 1 uL, Taqmix 12.5 uL, 16S upstream and downstream primers 1 uL, ddH2O10.5. mu.L. Reaction procedure: pre-denaturation at 98 deg.C for 10min, denaturation at 94 deg.C for 50s, annealing at 55 deg.C for 45s, extension at 72 deg.C for 1min, 30 cycles, extension at 72 deg.C for 10min, and storage at 4 deg.C. And (3) carrying out PCR band purification on the single band of the target gene obtained by amplification, carrying out gel cutting purification on the band with a non-specific amplification band. Carrying out Sanger sequencing on the obtained purified 5.8S rDNA, bringing the sequencing result into GeneBank for comparison, and determining the screened yeast species, wherein the upstream and downstream primers are as follows: upstream primer ITS1, SEQ ID NO: 5, 5'-tccgtaggtgaacctgcgg-3'; downstream primer ITS4, SEQ ID NO: 6, 5'-tcctccgcttattgatatgc-3'.
1.3.3 measurement of amino acids were measured according to the national standard method (GB 5009.124-2016).
2, separating and purifying the degrading bacteria. The 9 plant samples were screened according to the method of 1.3.1 to obtain 3 yeasts.
2.1 study of flavor-producing substances: three separated and purified yeasts are respectively inoculated into a 50mLYPD culture medium, cultured at 200r/min and 30 ℃ for 20h, fermentation liquor is taken to analyze glutamic acid, alanine and glycine, the variation of amino acid before and after fermentation is shown in table 3, and the variation condition of amino acid before and after fermentation is shown in table 3.
TABLE 3
2.2 the colony morphology identification of TM-3, the colony characteristics are as follows: the colony protrusions are white, the edges are regular, and the surface is smooth and wet, as shown in FIG. 5.
2.3.1 molecular characterization: the bacterial liquid is used as a template, 5.8S universal primers are used for gene amplification, PCR products are sequenced, and the sequencing results are compared in a GeneBank database, so that the homology between the strain TM-3 and the Saccharomyces cerevisiae STY2-1 is up to 100 percent. The result of the phylogenetic tree constructed by the MEGA-5.0 software for the TM-3 strain by the N-J method is shown in FIG. 6. According to the similarity, phylogenetic tree and colony morphology of the strain 5.8SrDNA, the TM-3 strain is preliminarily identified to be Saccharomyces cerevisiae (Saccharomyces cerevisiae), the Latin name of the Saccharomyces cerevisiae is Saccharomyces cerevisiae, the strain is preserved in China general microbiological culture Collection center of China Committee for culture Collection of microorganisms in 13 days 09 and 13 months in 2018, the preservation number is CGMCC NO.16469, and the preservation address is China institute of microbiology, Ministry of sciences, China, No. 3, North Kyowa Yang district, N.R. 1 of North Chen, China.
Example 4 screening of lactic acid bacteria with flavor-enhancing substances
1, materials and methods: the samples are respectively derived from a sour porridge sample, a pickled vegetable sample, a yoghurt sample and a corn by-product sample.
1.1 reagents and media: MRS culture medium: MRS pre-prepared medium 48g/L, solid medium added with 1.5% agar.
1.2 main instruments and equipment: SBA-40D type biosensing analyzer; a PCR instrument: BIO-RAD, USA; gel imager: BIO-RAD, USA;
1.3L-lactic acid determination method, taking 2mL of lactic acid fermentation liquor, centrifuging to remove calcium carbonate precipitate in the fermentation liquor, diluting the sample to a linear range, and waiting for determination. First, the instrument was cleaned automatically 2 times, after the electrodes were equilibrated, 25 μ L of the standard sample was aspirated for calibration, and the sample was measured after calibration was completed. And sucking 25 mu L of the supernatant liquid diluted by corresponding times by using a sample injection needle, injecting the supernatant liquid into the reaction cell, and reading on an instrument screen to obtain the determined yield of the L-lactic acid.
1.3.1 enrichment: 5g of the sample is taken and placed in 50mL of sterile MRS medium for static culture for 2d at 37 ℃. Screening: adding 10mL of each sample fermentation liquid obtained by enrichment into a triangular flask filled with 90mL of sterile water to obtain 10-1 diluent, shaking and uniformly mixing, sucking 1mL of the mixture, transferring the mixture into a test tube filled with 9mL of sterile water to prepare 10-2 diluent, sequentially preparing 10-3, 10-4, 10-5 and 10-6 diluents, sucking 0.2mL of 10-4, 10-5 and 10-6 diluents respectively, coating the diluents on an MRS solid culture medium, inverting the plate at 37 ℃ for culturing for 48 hours, and picking the lactic acid bacteria strain with the dominant growth quantity on the plate for further research.
1.3.2 morphological identification of lactic acid bacteria: the obtained strain is streaked on an MRS plate, cultured for 24-48h at 37 ℃, and observed for colony morphology, size, edge, surface bulge shape, transparency and the like. Gram staining of the strain was performed.
And (3) molecular identification: 16SrDNA identification is carried out on the obtained degradation bacteria, bacteria liquid or single colony is used as a template, and a reaction system is as follows: template 1 uL, Taqmix 12.5 uL, 16S upstream and downstream primers 1 uL, ddH2O10.5. mu.L. Reaction procedure: pre-denaturation at 98 deg.C for 10min, denaturation at 94 deg.C for 50s, annealing at 55 deg.C for 45s, extension at 72 deg.C for 1.5min, 30 cycles, extension at 72 deg.C for 10min, and storage at 4 deg.C. And detecting the PCR product through nucleic acid electrophoresis, sending the PCR product to Invitrogen company for sequencing, and comparing the sequence obtained by sequencing on NCBI. The sequence of the bacterial universal primer is shown as SEQ ID NO: 1 and SEQ ID NO: 2, respectively.
2, separating and purifying the degrading bacteria, and screening 7 samples according to the method of 1.3.1 to obtain 8 lactobacillus strains.
2.1 taking the 8 strains, selecting a single strain to be dropped into a 50mLMRS fermentation culture medium, taking a fermentation supernatant after anaerobic standing for 2d, and measuring the content of the lactic acid bacteria by the method in 1.3.1, wherein the results are shown in Table 4, the lactic acid producing capability of the lactic acid bacteria is shown in Table 4, and the lactic acid producing capability of the RS-5 strain is the highest. Therefore, RS-5 strain is selected for identification.
TABLE 4
2.2 morphological identification of RS-5 degrading bacteria, wherein the colony characteristics are as follows: the colonies were flat, regular in edge, smooth in surface, milky white, gram-positive, and the picture of the colonies is shown in FIG. 7.
2.2.1 molecular identification, using bacterial liquid as a template, using 16S universal primers to carry out gene amplification, sequencing PCR products, and comparing sequencing results in a GeneBank database, wherein results show that the homology of the strain RS-5 and the Lactobacillus plantarum Uruma-SU11 is as high as 99%. The result of constructing a phylogenetic tree of the RS-5 strain by the N-J method using MEGA-5.0 software is shown in FIG. 8. According to the similarity of the strain 16SrDNA, phylogenetic tree and colony morphology, the RS-5 strain is preliminarily identified as Lactobacillus plantarum, the Latin name of the Lactobacillus plantarum is Lactobacillus plantarum, the strain is preserved in China general microbiological culture Collection center in 2018, 08 and 14 days, the preservation number is CGMCC NO.16256, and the preservation address is China institute of sciences institute of microbiology, No. 3, North Chen Lu No.1 institute of China, Beijing Korean district, China.
Example 5 production of ZEN degrading enzyme by Pichia pastoris
The preparation method and process of the ZEN degrading enzyme and the verification content of the results of each process in the embodiment are disclosed in patent application No. 201110082679. X.
And adding 100L of water into 100L of fermentation liquor of the pichia pastoris GS115 expression strain for dilution, introducing the diluted fermentation liquor into a disc centrifuge for centrifugal sterilization, and rotating speed of 5000 r/min. And (3) introducing the sterilized fermentation liquor into a ceramic membrane filter for fine filtration and impurity removal, wherein the aperture of the ceramic membrane is 50nm, and the pressure is less than 0.6 Mpa. And (3) concentrating the liquid after fine filtration to 30L by an ultrafiltration membrane concentration device and an ultrafiltration membrane with the aperture of 50KDa to obtain yellow clear ZEN degrading enzyme liquid. Adding 10-20% of maltodextrin into the yellow ZEN degrading enzyme concentrated solution, fully stirring, and then carrying out spray drying at the air inlet temperature of 180-
Example 6 production of DON-degrading enzyme by Bacillus subtilis
1.1 Experimental materials: seed culture medium: 10g/L of peptone, 5g/L of yeast powder and 10g/L of NaCl; fermentation medium: 20g/L of corn flour, 20g/L of corn steep liquor and 20g/L of soybean meal.
1.2 Experimental flow:
(1) first-stage seed liquid preparation, selecting activated degraded DON bacillus subtilis single colony to 3.5L of sterile seed liquid, rotating speed of 220rpm, temperature of 37 ℃, and culturing for 16 h.
(2) Preparing a secondary seed liquid: adding 70L sterile seed culture medium into 100L fermentation tank, rotating speed 500r/min, temperature 37 deg.C, ventilating 0.5vvm, dissolved oxygen greater than 20%, and culturing for 12 h.
(3) Preparing for fermentation: 700L of corn steep liquor was added to a 1t fermenter, sterilized, and adjusted to pH7.0 with liquid ammonia.
(4) Fermentation: introducing the activated bacteria liquid into a 1t fermentation tank, rotating at a speed of 500r/min, at a temperature of 37 deg.C, ventilating at a pressure of 0.5vvm, and dissolving oxygen more than 20%
Preparation of 2DON degrading enzyme preparation: and adding 700L of water into the fermentation broth for dilution, and introducing the diluted fermentation broth into a disc centrifuge for centrifugal sterilization at the rotation speed of 5000 r/min. And (3) introducing the sterilized fermentation liquor into a ceramic membrane filter for fine filtration and impurity removal, wherein the aperture of the ceramic membrane is 50nm, and the pressure is less than 0.6 Mpa. And (3) concentrating the liquid after fine filtration to 30L by an ultrafiltration membrane concentration device and an ultrafiltration membrane with the aperture of 50KDa to obtain yellow clear DON degrading enzyme liquid. Adding 10-20% of maltodextrin into the yellow DON degrading enzyme concentrated solution, fully stirring, and then carrying out spray drying at the air inlet temperature of 180-.
Example 7 degradation of ZEN toxin in corn steep liquor by ZEN enzyme preparation
1. Experimental materials: corn bran: 300 kg; corn thick syrup (40% dry matter content): 400L; liquid ammonia; LB culture medium: 5g/L of yeast powder, 10g/L of peptone and 10g/L of NaCl; degrading ZEN enzyme preparation.
2. The experimental process comprises the following steps: preparing: adding 400L of concentrated corn pulp into a 1t fermentation tank, regulating the pH value to 7.0 by liquid ammonia at the rotation speed of 500r/min and the temperature of 37 ℃; enzymolysis: and (3) adding 220g of ZEN enzyme preparation powder into a fermentation tank, carrying out enzymolysis for 2h at the rotation speed of 500r/min and the pH value of 7.0 and at the temperature of 37 ℃ to obtain the ZEN detoxified corn steep liquor. And (3) spraying slurry: mixing concentrated detoxicated corn steep liquor with corn bran, drying in tube bundle drier and crushing; and bagging the crushed corn husks subjected to pulp spraying.
3. The mycotoxin assay in corn steep liquor is shown in table 5.
TABLE 5
As can be seen from Table 5, the ZEN toxin is degraded from initial 5704.52 mug/kg to 47.93 mug/kg, the degradation effect is good, and the standard of the mycotoxin in national feed is met.
Example 8 degradation of DON toxin in corn steep liquor by DON enzyme preparation
1. Experimental materials: corn bran: 300 kg; corn thick syrup (40% dry matter content): 400L; liquid ammonia; degrading DON enzyme preparation.
2. The experimental process comprises the following steps: preparing: adding 400L of concentrated corn pulp into a 1t fermentation tank, regulating the pH value to 7.0 by liquid ammonia at the rotation speed of 500r/min and the temperature of 37 ℃; enzymolysis: adding 220g of enzyme preparation powder into a fermentation tank, performing enzymolysis at a rotation speed of 500r/min and a pH value of 7.0 at a temperature of 37 ℃ for 2h to obtain DON detoxified corn steep liquor; and (3) spraying slurry: mixing concentrated detoxicated corn steep liquor with corn bran, drying in tube bundle drier and crushing; and bagging the crushed corn husks subjected to pulp spraying.
3. Mycotoxin assay in corn steep liquor is shown in Table 6
TABLE 6
As can be seen from the graph 6, before and after the sample treatment, DON toxin is degraded from initial 6834.9 mug/kg to 83.43 mug/kg, the degradation effect is good, and the DON toxin meets the mycotoxin standard in national feed.
Example 9 degradation of ZEN toxin in corn steep liquor by Aspergillus niger ZEN fermentation
1. Experimental materials: corn bran: 300 kg; corn thick syrup (20% dry matter content): 700L; liquid ammonia; activating a culture medium: molasses 20g/L, peptone 20g/L, KH2PO40.2g/L，MgSO4·7H2O 0.2g/L，(NH4)2HPO40.4g/L, tween-6015 g/L and corn flour 5 g/L; aspergillus niger.
2. The experimental process comprises the following steps: activating a microbial inoculum, adding 10L of activation culture medium into a 20L fermentation tank, adding 70g of Aspergillus niger spore reduction microbial inoculum, rotating at a speed of 500r/min, at a temperature of 37 ℃, ventilating at a speed of 0.5vvm, and activating for 2h when dissolved oxygen is more than 20%; preparing for fermentation: adding 700L of corn steep liquor into a 1t fermentation tank, sterilizing, and adjusting the pH value to 7.0 by using liquid ammonia; fermentation: introducing the activated bacterial liquid into a 1t fermentation tank, rotating at the speed of 500r/min, ventilating at the temperature of 37 ℃, and ventilating for 0.5vvm, wherein the dissolved oxygen is more than 20 percent, so as to obtain the ZEN detoxified corn steep liquor; concentrating corn steep liquor: putting the detoxified corn steep liquor into an evaporator, and evaporating and concentrating to 400L; and (3) spraying slurry: mixing concentrated detoxicated corn steep liquor with corn bran, drying in tube bundle drier and crushing; and bagging the crushed corn husks subjected to pulp spraying.
3. The mycotoxin assay in corn steep liquor is shown in table 7.
TABLE 7
As can be seen from Table 7, the ZEN toxin degraded from the initial 5629.43 μ g/kg to 145.61 μ g/kg. The degradation effect is good, and the mycotoxin standard in the national feed is met.
Example 10 degradation of DON toxin in corn steep liquor by fermentation of Bacillus subtilis DON
1. Experimental materials: corn bran: 300 kg; corn thick syrup (20% dry matter content): 700L; liquid ammonia; activating a culture medium: 5g/L of yeast powder, 10g/L of peptone and 10g/L of NaCl; a bacillus subtilis agent; saccharomyces cerevisiae.
2. The experimental process comprises the following steps: activating a microbial inoculum, adding 10L of activation culture medium into a 20L fermentation tank, adding 70g of DON-reducing bacillus subtilis microbial inoculum, rotating at the speed of 500r/min, at the temperature of 37 ℃, ventilating at 0.5vvm, and activating for 2h when the dissolved oxygen is more than 20%; preparing for fermentation: adding 700L of corn steep liquor into a 1t fermentation tank, sterilizing, and adjusting the pH value to 7.0 by using liquid ammonia; fermentation: introducing the activated bacterial liquid into a 1t fermentation tank, rotating at a speed of 500r/min, ventilating at a temperature of 37 ℃, and ventilating at a pressure of 0.5vvm, wherein the dissolved oxygen is more than 20 percent, so as to obtain DON detoxified corn steep liquor; concentrating corn steep liquor: putting the detoxified corn steep liquor into an evaporator, and evaporating and concentrating to 400L; and (3) spraying slurry: mixing concentrated detoxicated corn steep liquor with corn bran, drying in tube bundle drier and crushing; and bagging the crushed corn husks subjected to pulp spraying.
3. The mycotoxin assay in corn steep liquor is shown in table 8.
TABLE 8
As can be seen from Table 8, the DON toxin degraded from the initial 6734.9 μ g/kg to 107.11 μ g/kg. The degradation effect is good, and the mycotoxin standard in the national feed is met.
Example 11 Aspergillus niger ZEN fermentation degrades ZEN toxin in corn steep liquor and DON enzyme preparation degrades DON toxin
1. Experimental materials: corn bran: 300 kg; corn thick syrup (20% dry matter content): 700L; liquid ammonia; activating a culture medium: molasses 20g/L, peptone 20g/L, KH2PO40.2g/L，MgSO4·7H2O 0.2g/L，(NH4)2HPO40.4g/L, Tween-6015 g/L, corn flour 5 g/L; degrading DON enzyme preparation; degrading ZEN Aspergillus niger microbial inoculum.
2. The experimental process comprises the following steps: activating a microbial inoculum, adding 10L of activation medium into a 20L fermentation tank, adding 70g of ZEN-reducing Aspergillus niger microbial inoculum, rotating at a speed of 500r/min, at a temperature of 37 ℃, ventilating at 0.5vvm, and activating for 2h when the dissolved oxygen is more than 20%; preparing for fermentation, adding 700L of corn steep liquor into a 1t fermentation tank, sterilizing, and adjusting pH to 7.0 with liquid ammonia; fermentation: introducing the activated bacterial liquid into a 1t fermentation tank, wherein the rotating speed is 500r/min, the temperature is 37 ℃, the ventilation is 0.5vvm, and the dissolved oxygen is more than 20%; carrying out enzymolysis, namely adding 220g of DON degrading enzyme preparation powder into a fermentation tank after fermenting and culturing for 18h, and carrying out enzymolysis for 2h at the rotation speed of 500r/min, the pH value of 7.0 and the temperature of 37 ℃ to obtain ZEN and DON detoxified corn steep liquor; concentrating corn steep liquor: putting the detoxified corn steep liquor into an evaporator, and evaporating and concentrating to 400L; and (3) spraying slurry: mixing concentrated detoxicated corn steep liquor with corn bran, drying in tube bundle drier and crushing; bagging the crushed corn bran which is sprayed with slurry
3. Mycotoxin assay in corn steep liquor as shown in Table 9
TABLE 9
As can be seen from Table 9, before and after sample treatment, the ZEN toxin degraded from the initial 5131.28 μ g/kg to 74.15 μ g/kg, and the DON toxin degraded from the initial 6362.42 μ g/kg to 42.94 μ g/kg. The degradation effect is good, and the mycotoxin standard in the national feed is met.
Example 12 degradation of ZEN toxin by ZEN enzyme preparation and degradation of DON toxin in corn steep liquor by fermentation of Bacillus subtilis
1. Experimental materials: corn bran: 300 kg; corn thick syrup (20% dry matter content): 700L; liquid ammonia; activating a culture medium: 5g/L of yeast powder, 10g/L of peptone and 10g/L of NaCl; degrading the ZEN enzyme preparation; degrading DON bacillus subtilis microbial inoculum.
2. The experimental process comprises the following steps: activating a microbial inoculum, adding 10L of activation culture medium into a 20L fermentation tank, adding 70g of DON-reducing bacillus subtilis microbial inoculum, rotating at the speed of 500r/min, at the temperature of 37 ℃, ventilating at 0.5vvm, and activating for 2h when the dissolved oxygen is more than 20%; preparing for fermentation, adding 700L of corn steep liquor into a 1t fermentation tank, sterilizing, and adjusting pH to 7.0 with liquid ammonia; introducing the activated bacterial liquid into a 1t fermentation tank, wherein the rotating speed is 500r/min, the temperature is 37 ℃, the ventilation is 0.5vvm, and the dissolved oxygen is more than 20%; carrying out enzymolysis, namely adding 220g of enzyme preparation powder into a fermentation tank after fermenting and culturing for 18h, and carrying out enzymolysis for 2h at the rotation speed of 500r/min, the pH value of 7.0 and the temperature of 37 ℃ to obtain ZEN and DON detoxified corn steep liquor; concentrating the corn steep liquor, namely putting the detoxified corn steep liquor into an evaporator, and evaporating and concentrating to 400L; mixing concentrated detoxicated corn steep liquor with corn bran, drying in tube bundle drier and crushing; and bagging the crushed corn husks subjected to pulp spraying.
3. Mycotoxin assay in corn steep liquor as shown in Table 10
Watch 10
From Table 10, before and after sample treatment, the ZEN toxin degraded from the initial 5289.74 μ g/kg to 22.42 μ g/kg and the DON toxin degraded from the initial 6062.21 μ g/kg to 72.34 μ g/kg. The degradation effect is good, and the mycotoxin standard in the national feed is met.
Example 13 degradation of ZEN and DON, Yeast and lactic acid bacteria flavor enhancement in corn steep liquor by fermentation of Bacillus subtilis and Aspergillus niger
1. Experimental materials: corn bran: 300 kg; corn thick syrup (20% dry matter content): 700L; liquid ammonia; activating a culture medium: molasses 20g/L, peptone 20g/L, KH2PO40.2g/L，MgSO4·7H2O 0.2g/L，(NH4)2HPO40. L, Tween-6015 g/L, corn flour 5 g/L; aspergillus niger, Bacillus subtilis, Saccharomyces cerevisiae and lactobacillus.
2. The experimental process comprises the following steps: activating a microbial inoculum, adding 10L of an activation culture medium into a 20L fermentation tank, adding 70g of Aspergillus niger spore reduction microbial inoculum, 70g of Bacillus subtilis microbial inoculum and 100g of Saccharomyces cerevisiae microbial inoculum, rotating at 500r/min, ventilating at 37 ℃, ventilating at 0.5vvm, and activating for 2h when the dissolved oxygen is more than 20%; preparing for fermentation, adding 700L of corn steep liquor into a 1t fermentation tank, sterilizing, and adjusting pH to 7.0 with liquid ammonia; introducing the activated bacterial liquid into a 1t fermentation tank, rotating at the speed of 500r/min, ventilating at the temperature of 37 ℃, and ventilating at 0.5vvm, wherein the dissolved oxygen is more than 20%, so as to obtain flavor ZEN and DON detoxified corn steep liquor; concentrating the corn steep liquor, namely putting the detoxified corn steep liquor into an evaporator, and evaporating and concentrating to 400L; mixing concentrated detoxicated corn steep liquor with corn bran, drying in tube bundle drier and crushing; bagging the crushed corn bran which is sprayed with slurry
3. Mycotoxin assay in corn steep liquor is shown in table 11.
TABLE 11
The flavor substances in the corn steep liquor were measured as shown in table 12.
TABLE 12
As can be seen from Table 11, the ZEN toxin degraded from the initial 5421.36 μ g/kg to 121.82 μ g/kg, and the DON toxin degraded from the initial 6346.92 μ g/kg to 163.82 μ g/kg. The degradation effect is good, and the mycotoxin standard in the national feed is met. As shown in Table 12, the water-soluble protein, L-lactic acid, L-sodium glutamate, L-alanine and L-glycine of the gunite corn before and after fermentation were detected, and the indexes were significantly increased. The addition of the water-soluble protein enables the sprayed corn bran to be used as animal feed and is more beneficial to the absorption of nutrition, the obvious addition of the lactic acid increases sour substances in the sprayed corn bran and improves the palatability of the sprayed corn bran, and the L-sodium glutamate, the L-alanine and the L-glycine can relieve the alkaline taste in the sprayed corn bran and improve the fragrance in the sprayed corn bran, so that the fermented sprayed corn bran has better feeding attraction and is also beneficial to the absorption of nutrient substances such as protein.
Example 14 Yeast, Lactobacillus-flavor enhancing substance, ZEN, DON enzyme preparation degradation of ZEN and DON toxins
1. Experimental materials: corn bran: 300 kg; corn thick syrup (20% dry matter content): 700L; liquid ammonia; activating a culture medium: molasses 20g/L, peptone 20g/L, KH2PO40.2g/L，MgSO4·7H2O 0.2g/L，(NH4)2HPO40.4g/L, tween-6015 g/L and corn flour 5 g/L; saccharomyces cerevisiae microbial inoculum, lactobacillus microbial inoculum, and enzyme preparation for degrading ZEN and DON.
2. The experimental process comprises the following steps: activating a microbial inoculum, adding 10L of activation medium into a 20L fermentation tank, adding 100g of saccharomyces cerevisiae microbial inoculum, rotating speed of 100g of lactic acid microbial inoculum is 500r/min, temperature is 37 ℃, ventilation is 0.5vvm, dissolved oxygen is more than 20%, and activating for 2 h; preparing for fermentation: adding 700L of corn steep liquor into a 1t fermentation tank, sterilizing, and adjusting the pH value to 7.0 by using liquid ammonia; fermenting, namely introducing the activated bacterium liquid into a 1t fermentation tank, wherein the rotating speed is 500r/min, the temperature is 37 ℃, the ventilation is 0.5vvm, and the dissolved oxygen is more than 20%; concentrating the corn steep liquor, namely putting the detoxified corn steep liquor into an evaporator, and evaporating and concentrating to 400L; carrying out enzymolysis, namely adding 220g of degraded DON enzyme preparation powder and 220g of degraded ZEN enzyme preparation powder into a fermentation tank, and carrying out enzymolysis for 2 hours at the rotating speed of 500r/min, the pH value of 7.0 and the temperature of 37 ℃ to obtain flavored ZEN and DON detoxified corn steep liquor; spraying slurry, namely adding concentrated detoxified corn slurry into a flow to mix with corn bran, drying by a tube bundle dryer and then crushing; and bagging the crushed corn husks subjected to pulp spraying.
3. Mycotoxin assay in corn steep liquor is shown in table 13.
Watch 13
The detection of flavor substances in the corn steep liquor is shown in Table 14.
TABLE 14
As shown in Table 13, the ZEN toxin degraded from 5441.78 μ g/kg initially to 70.02 μ g/kg, and the DON toxin degraded from 6306.22 μ g/kg initially to 34.12 μ g/kg. The degradation effect is good, and the mycotoxin standard in the national feed is met. As shown in Table 14, the water-soluble protein, L-lactic acid, L-sodium glutamate, L-alanine and L-glycine of the shotcrete corn before and after fermentation were detected, and the indexes were significantly increased. The addition of the water-soluble protein enables the sprayed corn bran to be used as animal feed and is more beneficial to the absorption of nutrition, the obvious addition of the lactic acid increases sour substances in the sprayed corn bran and improves the palatability of the sprayed corn bran, and the L-sodium glutamate, the L-alanine and the L-glycine can relieve the alkaline taste in the sprayed corn bran and improve the fragrance in the sprayed corn bran, so that the fermented sprayed corn bran has better feeding attraction and is also beneficial to the absorption of nutrient substances such as protein.
Example 15 preparation of solid fermentation flavored jet corn bran
1. Experimental materials: corn bran: 300 kg; corn thick syrup (40% dry matter content): 400L; liquid ammonia; activating a culture medium: molasses 20g/L, peptone 20g/L, KH2PO40.2g/L，MgSO4·7H2O 0.2g/L，(NH4)2HPO40.4g/L, tween-6015 g/L and corn flour 5 g/L; aspergillus niger agent, Bacillus subtilis agent; a saccharomyces cerevisiae agent; a lactic acid bacteria agent.
2. The experimental process comprises the following steps: activating a microbial inoculum, adding 10L of an activation culture medium into a 20L fermentation tank, adding 70g of Aspergillus niger spore reduction microbial inoculum, 70g of Bacillus subtilis microbial inoculum and 100g of Saccharomyces cerevisiae microbial inoculum, rotating at 500r/min, ventilating at 37 ℃, ventilating at 0.5vvm, and activating for 2h when the dissolved oxygen is more than 20%; preparing for fermentation: mixing 300kg of corn husks and 400L of corn steep liquor in a feed mixer, and adjusting the pH value to 7.0 by using liquid ammonia; inoculation: introducing the activated bacterial liquid into a feed mixer, and fully stirring the materials; fermentation: putting the inoculated gunite corn bran into a fermentation bag, sealing the fermentation bag, and standing for fermentation for 4 days; drying and crushing the fermented gunite corn husk tube bundle dryer; and bagging the crushed corn husks subjected to pulp spraying.
3. The mycotoxin assay in corn steep liquor is shown in table 15.
Watch 15
The detection of flavor substances in the corn steep liquor is shown in table 16.
TABLE 16
As shown in Table 15, the ZEN toxin degraded from the initial 5221.36 μ g/kg to 820.32 μ g/kg and the DON toxin degraded from the initial 6246.81 μ g/kg to 1063.23 μ g/kg. The degradation effect is good, and the mycotoxin standard in the national feed is met. As shown in table 16, by detecting L-lactic acid, L-sodium glutamate, L-alanine and L-glycine in the gunite corn bran before and after fermentation, each index is significantly increased, so that the gunite corn bran is used as an animal feed and is more beneficial to the absorption of nutrition, the lactic acid is significantly increased, sour substances in the gunite corn bran are increased, the palatability of the gunite corn bran is improved, and the L-sodium glutamate, the L-alanine and the L-glycine can all relieve the alkaline taste in the gunite corn bran, improve the fragrance in the gunite corn bran, make the fermented gunite corn bran more attractive, and are also beneficial to the absorption of nutritional substances such as protein.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Sequence listing
<110> national institute of food and Material Reserve science
<120> preparation method of flavor detoxified corn steep liquor
<130>GG19624424A
<160>6
<170>SIPOSequenceListing 1.0
<210>1
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>1
attgaacgct ggcggcaggc ct 22
<210>2
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>2
taacaaggta accgtagggg a 21
<210>3
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>3
gtagtcatat gcttgtctc 19
<210>4
<211>18
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>4
attccccgtt acccgttg 18
<210>5
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>5
tccgtaggtg aacctgcgg 19
<210>6
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>6
tcctccgctt attgatatgc 20
Claims (10)
1. A preparation method of flavor detoxified corn steep liquor, which is characterized in that,
adding DON degrading bacteria or degrading enzyme into the corn steep liquor, and degrading DON toxin by a biological method;
adding ZEN degrading bacteria or degrading enzyme into the corn steep liquor, and degrading ZEN toxin by a biological method;
adding lactobacillus and yeast into the corn steep liquor for fermentation, improving the content of amino acid and lactic acid in the corn steep liquor, and improving the flavor of the corn steep liquor to obtain the flavor-detoxified corn steep liquor.
2. The method of making flavor-detoxified corn steep liquor of claim 1,
the DON degrading bacteria are Bacillus subtilis with the Latin name of Bacillus subtilis, the Bacillus subtilis is preserved in China general microbiological culture Collection center in 2018, 08 and 13 months, the preservation number is CGMCC NO.16254, and the preservation address is as follows: the institute of microbiology, national academy of sciences No. 3, Xilu No.1, Beijing Korean district, China;
the ZEN degrading bacteria are Aspergillus niger, the Latin name of the ZEN degrading bacteria is Aspergillus niger, the ZEN degrading bacteria are preserved in China general microbiological culture Collection center (CGMCC) in 11-23 th of 2017, the preservation number is CGMCC NO.14639, and the preservation address is as follows: the institute of microbiology, China academy of sciences, No. 3, Xilu No.1, Beijing Korean district, Beichen, China.
3. The method of making flavor-detoxified corn steep liquor of claim 1,
the lactobacillus is lactobacillus plantarum, the Latin name of the lactobacillus plantarum is Lactobacillus plantarum, the lactobacillus is preserved in the China general microbiological culture Collection center in 2018, 08 and 14 months, the preservation number is CGMCC No.16256, and the preservation address is as follows: the institute of microbiology, national academy of sciences No. 3, Xilu No.1, Beijing Korean district, China;
the yeast is Saccharomyces cerevisiae, the Latin name of the Saccharomyces cerevisiae is Saccharomyces cerevisiae, the strain is preserved in China general microbiological culture Collection center in 13.09.2018, the preservation number is CGMCC No.16469, and the preservation address is as follows: the institute of microbiology, China academy of sciences, No. 3, Xilu No.1, Beijing Korean district, Beichen, China.
4. The method of making flavor-detoxified corn steep liquor of claim 1,
adding ZEN degrading bacteria or ZEN degrading enzyme into the corn steep liquor, and adding DON degrading bacteria or DON enzyme for degradation specifically comprises the following steps:
adding any one of ZEN degrading bacteria and DON degrading bacteria, ZEN degrading bacteria and DON degrading enzyme, ZEN degrading enzyme and DON degrading bacteria, ZEN degrading enzyme and DON degrading enzyme, ZEN degrading bacteria and DON degrading enzyme into the corn steep liquor, wherein ZEN degrading and DON degrading can be carried out step by step or synchronously.
5. The method of making flavor-detoxified corn steep liquor of claim 1,
the ZEN content in the flavor detoxified corn steep liquor is lower than 100 mug/kg, the DON content is lower than 500 mug/kg, and the probiotic content is more than or equal to 109CFU/ml, increased lactic acid, glutamic acid and glycine content.
6. The method of making flavor-detoxified corn steep liquor of claim 1,
the ZEN and DON degrading enzyme is a solid enzyme preparation or a liquid enzyme preparation;
the ZEN and DON degrading bacteria are solid bacteria preparation or liquid activated seed liquid.
7. The method of making flavor-detoxified corn steep liquor of claim 6,
the activation method of the ZEN and DON degrading bacteria is characterized in that the ZEN and DON degrading bacteria are respectively inoculated into a fermentation tank containing an activation culture medium and activated for 2-3 hours to prepare the ZEN and DON degrading bacteria;
wherein the rotating speed of the fermentation tank is 100-.
8. The method of making flavor-detoxified corn steep liquor of claim 1,
the degrading enzyme is prepared by fermenting degrading bacteria or is introduced into host cells to be a recombinant expression vector with ZEN degrading enzyme coding genes, and then the ZEN and DON degrading enzyme is obtained by host cell expression.
9. A preparation method of a flavor feed product is characterized in that,
spraying the flavored detoxified corn steep liquor prepared by the method of claim 1 onto a feed product.
10. The method of claim 9, wherein the step of,
the feed product comprises corn bran, wheat bran or corn germ meal.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911194779.4A CN111011707A (en) | 2019-11-28 | 2019-11-28 | Preparation method of flavor detoxified corn steep liquor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911194779.4A CN111011707A (en) | 2019-11-28 | 2019-11-28 | Preparation method of flavor detoxified corn steep liquor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111011707A true CN111011707A (en) | 2020-04-17 |
Family
ID=70203262
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911194779.4A Pending CN111011707A (en) | 2019-11-28 | 2019-11-28 | Preparation method of flavor detoxified corn steep liquor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111011707A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117243296A (en) * | 2023-10-13 | 2023-12-19 | 山东星光首创生物科技有限公司 | Method for preparing gunite corn husks by recycling waste saccharomycetes in erythritol production |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009167138A (en) * | 2008-01-18 | 2009-07-30 | Univ Of Tokyo | Mycotoxin production inhibitor |
| CN102827881A (en) * | 2012-09-11 | 2012-12-19 | 国家粮食局科学研究院 | Biological degradation method for fungaltoxin in grain and/or grain byproducts |
| CN103829036A (en) * | 2014-02-17 | 2014-06-04 | 郑州市中食农产品加工研究院 | Preparation method of microecological fermented feed by utilizing byproducts of corn deep-processing as raw materials |
| CN104099251A (en) * | 2014-04-11 | 2014-10-15 | 江南大学 | New aspergillus niger strain and application thereof in degradation of a plurality of kinds of fungaltoxin |
| CN106509590A (en) * | 2016-10-26 | 2017-03-22 | 北京科润生科技发展有限公司 | Detoxification agent for vomitoxin in biodegradable feed and preparation technology thereof |
| CN109043299A (en) * | 2018-07-03 | 2018-12-21 | 山东寿光巨能金玉米开发有限公司 | A method of utilizing toxin in compound microorganism ferments method removing corn byproducts |
-
2019
- 2019-11-28 CN CN201911194779.4A patent/CN111011707A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009167138A (en) * | 2008-01-18 | 2009-07-30 | Univ Of Tokyo | Mycotoxin production inhibitor |
| CN102827881A (en) * | 2012-09-11 | 2012-12-19 | 国家粮食局科学研究院 | Biological degradation method for fungaltoxin in grain and/or grain byproducts |
| CN103829036A (en) * | 2014-02-17 | 2014-06-04 | 郑州市中食农产品加工研究院 | Preparation method of microecological fermented feed by utilizing byproducts of corn deep-processing as raw materials |
| CN104099251A (en) * | 2014-04-11 | 2014-10-15 | 江南大学 | New aspergillus niger strain and application thereof in degradation of a plurality of kinds of fungaltoxin |
| CN106509590A (en) * | 2016-10-26 | 2017-03-22 | 北京科润生科技发展有限公司 | Detoxification agent for vomitoxin in biodegradable feed and preparation technology thereof |
| CN109043299A (en) * | 2018-07-03 | 2018-12-21 | 山东寿光巨能金玉米开发有限公司 | A method of utilizing toxin in compound microorganism ferments method removing corn byproducts |
Non-Patent Citations (2)
| Title |
|---|
| 典姣姣等: "喷浆玉米皮混菌固态发酵饲料的研究", 《饲料研究》 * |
| 呙于明: "《动物营养研究进展2016》", 31 October 2016, 北京：中国农业大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117243296A (en) * | 2023-10-13 | 2023-12-19 | 山东星光首创生物科技有限公司 | Method for preparing gunite corn husks by recycling waste saccharomycetes in erythritol production |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104388514B (en) | The method that gamma aminobutyric acid is prepared using composite bacteria fermentation | |
| CN111826295B (en) | Bacillus belgii strain for fermenting soybean meal | |
| US8202710B2 (en) | Lactic acid bacterium capable of producing Y-aminobutyric acid | |
| CN114703096B (en) | Bacillus bailii strain, fermented feed degradation microbial toxin thereof and application | |
| CN110591943B (en) | Bacillus subtilis capable of producing complex enzyme, composition and application thereof | |
| CN113549574B (en) | Bacillus coagulans and application thereof | |
| CN110042072B (en) | Aflatoxin B degradation1And application thereof | |
| CN115948305B (en) | Bacillus subtilis capable of efficiently degrading various mycotoxins and application thereof | |
| CN110607251A (en) | Salt-tolerant marinobacter capable of degrading vomitoxin and application thereof | |
| CN116478891B (en) | Bacillus subtilis, microbial inoculum, fermented feed and preparation method thereof | |
| CN108034615A (en) | A kind of fermentation process of bacillus flow feeding | |
| CN113897319A (en) | Bacillus amyloliquefaciens and application thereof | |
| CN110878265A (en) | Bacillus subtilis for degrading aflatoxin and application thereof | |
| CN114107113B (en) | Method for reducing ethyl carbamate in fermented food by using synthetic starter | |
| CN106754507B (en) | Compound flavor microbial inoculum, preparation method thereof and direct-throwing application thereof in soy sauce flavoring | |
| CN113892556B (en) | Biological protein feed prepared from waste distillers' grains of Jimo old wine and preparation method thereof | |
| CN109423466B (en) | Compound fermentation inoculant and application thereof | |
| CN111011707A (en) | Preparation method of flavor detoxified corn steep liquor | |
| CN116064281B (en) | Bacillus bailii and application thereof in low-salt fermentation of food | |
| CN111040969B (en) | Compound lactobacillus agent and application thereof in buffalo silage | |
| CN115838643A (en) | Tea tree endophytic fungus and application thereof in promoting growth of dendrobium officinale | |
| CN115927032A (en) | Bacillus subtilis, leaven, preparation method and application thereof | |
| CN116004423A (en) | Bacillus bailii and application thereof | |
| CN114456990A (en) | Preparation method of DDGS (distillers dried grains with soluble) as well as fermentation strain and culture medium thereof | |
| CN110140810B (en) | Preparation method of fermented yellow wine lees for feed and fermented yellow wine lees |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200417 |
|
| RJ01 | Rejection of invention patent application after publication |