CN112574905A - 一种德氏乳杆菌及利用德氏乳杆菌发酵得到的豆乳饮料 - Google Patents
一种德氏乳杆菌及利用德氏乳杆菌发酵得到的豆乳饮料 Download PDFInfo
- Publication number
- CN112574905A CN112574905A CN202011205568.9A CN202011205568A CN112574905A CN 112574905 A CN112574905 A CN 112574905A CN 202011205568 A CN202011205568 A CN 202011205568A CN 112574905 A CN112574905 A CN 112574905A
- Authority
- CN
- China
- Prior art keywords
- lactobacillus delbrueckii
- soybean milk
- soybean
- fermentation
- isoflavone
- 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.)
- Granted
Links
- 235000010469 Glycine max Nutrition 0.000 title claims abstract description 80
- 244000068988 Glycine max Species 0.000 title claims abstract description 80
- 241000186673 Lactobacillus delbrueckii Species 0.000 title claims abstract description 71
- 235000020124 milk-based beverage Nutrition 0.000 title claims abstract description 8
- 235000013336 milk Nutrition 0.000 claims abstract description 44
- 239000008267 milk Substances 0.000 claims abstract description 44
- 210000004080 milk Anatomy 0.000 claims abstract description 44
- 239000000796 flavoring agent Substances 0.000 claims abstract description 13
- 235000019634 flavors Nutrition 0.000 claims abstract description 13
- 238000004321 preservation Methods 0.000 claims abstract description 10
- 238000000855 fermentation Methods 0.000 claims description 35
- 230000004151 fermentation Effects 0.000 claims description 35
- ZQSIJRDFPHDXIC-UHFFFAOYSA-N Daidzein Natural products C1=CC(O)=CC=C1C1=COC2=CC(O)=CC=C2C1=O ZQSIJRDFPHDXIC-UHFFFAOYSA-N 0.000 claims description 27
- ZCOLJUOHXJRHDI-CMWLGVBASA-N genistein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 ZCOLJUOHXJRHDI-CMWLGVBASA-N 0.000 claims description 21
- ZCOLJUOHXJRHDI-FZHKGVQDSA-N Genistein 7-O-glucoside Natural products O([C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1)c1cc(O)c2C(=O)C(c3ccc(O)cc3)=COc2c1 ZCOLJUOHXJRHDI-FZHKGVQDSA-N 0.000 claims description 16
- CJPNHKPXZYYCME-UHFFFAOYSA-N Genistin Natural products OCC1OC(Oc2ccc(O)c3OC(=CC(=O)c23)c4ccc(O)cc4)C(O)C(O)C1O CJPNHKPXZYYCME-UHFFFAOYSA-N 0.000 claims description 16
- YCUNGEJJOMKCGZ-UHFFFAOYSA-N Pallidiflorin Natural products C1=CC(OC)=CC=C1C1=COC2=CC=CC(O)=C2C1=O YCUNGEJJOMKCGZ-UHFFFAOYSA-N 0.000 claims description 16
- 102000006995 beta-Glucosidase Human genes 0.000 claims description 16
- 108010047754 beta-Glucosidase Proteins 0.000 claims description 16
- GMTUGPYJRUMVTC-UHFFFAOYSA-N Daidzin Natural products OC(COc1ccc2C(=O)C(=COc2c1)c3ccc(O)cc3)C(O)C(O)C(O)C=O GMTUGPYJRUMVTC-UHFFFAOYSA-N 0.000 claims description 13
- KYQZWONCHDNPDP-UHFFFAOYSA-N Daidzoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 KYQZWONCHDNPDP-UHFFFAOYSA-N 0.000 claims description 13
- KYQZWONCHDNPDP-QNDFHXLGSA-N daidzein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 KYQZWONCHDNPDP-QNDFHXLGSA-N 0.000 claims description 13
- 230000000694 effects Effects 0.000 claims description 10
- 235000008466 glycitein Nutrition 0.000 claims description 8
- NNUVCMKMNCKPKN-UHFFFAOYSA-N glycitein Natural products COc1c(O)ccc2OC=C(C(=O)c12)c3ccc(O)cc3 NNUVCMKMNCKPKN-UHFFFAOYSA-N 0.000 claims description 8
- DXYUAIFZCFRPTH-UHFFFAOYSA-N glycitein Chemical compound C1=C(O)C(OC)=CC(C2=O)=C1OC=C2C1=CC=C(O)C=C1 DXYUAIFZCFRPTH-UHFFFAOYSA-N 0.000 claims description 8
- 235000003687 soy isoflavones Nutrition 0.000 claims description 8
- 229930182470 glycoside Natural products 0.000 claims description 6
- 150000002338 glycosides Chemical class 0.000 claims description 5
- 230000003064 anti-oxidating effect Effects 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 3
- 230000009466 transformation Effects 0.000 claims description 3
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 abstract description 27
- 235000008696 isoflavones Nutrition 0.000 abstract description 27
- 238000006243 chemical reaction Methods 0.000 abstract description 26
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 abstract description 23
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 abstract description 8
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 abstract description 8
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 abstract description 8
- 238000004519 manufacturing process Methods 0.000 abstract description 8
- 230000003078 antioxidant effect Effects 0.000 abstract description 7
- 239000003963 antioxidant agent Substances 0.000 abstract description 6
- 235000006708 antioxidants Nutrition 0.000 abstract description 6
- 239000003205 fragrance Substances 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 2
- 230000003647 oxidation Effects 0.000 abstract 1
- 238000007254 oxidation reaction Methods 0.000 abstract 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 21
- 239000000243 solution Substances 0.000 description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 19
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 15
- 238000002474 experimental method Methods 0.000 description 12
- 235000013322 soy milk Nutrition 0.000 description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- ZWSNUPOSLDAWJS-QNDFHXLGSA-N 6,7-dihydroxy-3-[4-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]chromen-4-one Chemical compound OC[C@H]1O[C@@H](Oc2ccc(cc2)-c2coc3cc(O)c(O)cc3c2=O)[C@H](O)[C@@H](O)[C@@H]1O ZWSNUPOSLDAWJS-QNDFHXLGSA-N 0.000 description 8
- 238000009630 liquid culture Methods 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 241000186660 Lactobacillus Species 0.000 description 7
- 239000002253 acid Substances 0.000 description 7
- 230000001580 bacterial effect Effects 0.000 description 7
- 235000007240 daidzein Nutrition 0.000 description 7
- 239000012153 distilled water Substances 0.000 description 7
- 229940039696 lactobacillus Drugs 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- OZBAVEKZGSOMOJ-MIUGBVLSSA-N glycitin Chemical compound COC1=CC(C(C(C=2C=CC(O)=CC=2)=CO2)=O)=C2C=C1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O OZBAVEKZGSOMOJ-MIUGBVLSSA-N 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- 238000011156 evaluation Methods 0.000 description 5
- 235000006539 genistein Nutrition 0.000 description 5
- TZBJGXHYKVUXJN-UHFFFAOYSA-N genistein Natural products C1=CC(O)=CC=C1C1=COC2=CC(O)=CC(O)=C2C1=O TZBJGXHYKVUXJN-UHFFFAOYSA-N 0.000 description 5
- 229940045109 genistein Drugs 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000003752 polymerase chain reaction Methods 0.000 description 5
- 230000001953 sensory effect Effects 0.000 description 5
- 239000012086 standard solution Substances 0.000 description 5
- 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 4
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- JARKCYVAAOWBJS-UHFFFAOYSA-N hexanal Chemical compound CCCCCC=O JARKCYVAAOWBJS-UHFFFAOYSA-N 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- VSMOENVRRABVKN-UHFFFAOYSA-N oct-1-en-3-ol Chemical compound CCCCCC(O)C=C VSMOENVRRABVKN-UHFFFAOYSA-N 0.000 description 4
- KJFMBFZCATUALV-UHFFFAOYSA-N phenolphthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2C(=O)O1 KJFMBFZCATUALV-UHFFFAOYSA-N 0.000 description 4
- 239000006041 probiotic Substances 0.000 description 4
- 235000018291 probiotics Nutrition 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- -1 superoxide radicals Chemical class 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- XJTZHGNBKZYODI-UHFFFAOYSA-N Glycitin Natural products OCC1OC(Oc2ccc3OC=C(C(=O)c3c2CO)c4ccc(O)cc4)C(O)C(O)C1O XJTZHGNBKZYODI-UHFFFAOYSA-N 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 235000013365 dairy product Nutrition 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 239000000262 estrogen Substances 0.000 description 3
- 229940011871 estrogen Drugs 0.000 description 3
- 229930182478 glucoside Natural products 0.000 description 3
- 150000008131 glucosides Chemical class 0.000 description 3
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 3
- 150000002515 isoflavone derivatives Chemical class 0.000 description 3
- 230000031700 light absorption Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000008055 phosphate buffer solution Substances 0.000 description 3
- BUCIWTBCUUHRHZ-UHFFFAOYSA-K potassium;disodium;dihydrogen phosphate;hydrogen phosphate Chemical compound [Na+].[Na+].[K+].OP(O)([O-])=O.OP([O-])([O-])=O BUCIWTBCUUHRHZ-UHFFFAOYSA-K 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 description 3
- 239000008223 sterile water Substances 0.000 description 3
- 238000004448 titration Methods 0.000 description 3
- 235000013618 yogurt Nutrition 0.000 description 3
- MBDOYVRWFFCFHM-SNAWJCMRSA-N (2E)-hexenal Chemical compound CCC\C=C\C=O MBDOYVRWFFCFHM-SNAWJCMRSA-N 0.000 description 2
- JZQKTMZYLHNFPL-BLHCBFLLSA-N (2E,4E)-deca-2,4-dienal Chemical compound CCCCC\C=C\C=C\C=O JZQKTMZYLHNFPL-BLHCBFLLSA-N 0.000 description 2
- VSMOENVRRABVKN-MRVPVSSYSA-N 1-Octen-3-ol Natural products CCCCC[C@H](O)C=C VSMOENVRRABVKN-MRVPVSSYSA-N 0.000 description 2
- JZQKTMZYLHNFPL-UHFFFAOYSA-N 2-trans-4-trans-decadienal Natural products CCCCCC=CC=CC=O JZQKTMZYLHNFPL-UHFFFAOYSA-N 0.000 description 2
- YDXQPTHHAPCTPP-UHFFFAOYSA-N 3-Octen-1-ol Natural products CCCCC=CCCO YDXQPTHHAPCTPP-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 102000016938 Catalase Human genes 0.000 description 2
- 108010053835 Catalase Proteins 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- QSJXEFYPDANLFS-UHFFFAOYSA-N Diacetyl Chemical compound CC(=O)C(C)=O QSJXEFYPDANLFS-UHFFFAOYSA-N 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- AMQJEAYHLZJPGS-UHFFFAOYSA-N N-Pentanol Chemical compound CCCCCO AMQJEAYHLZJPGS-UHFFFAOYSA-N 0.000 description 2
- RQBBFKINEJYDOB-UHFFFAOYSA-N acetic acid;acetonitrile Chemical compound CC#N.CC(O)=O RQBBFKINEJYDOB-UHFFFAOYSA-N 0.000 description 2
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 229960005309 estradiol Drugs 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical compound CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000003301 hydrolyzing effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- TZMFJUDUGYTVRY-UHFFFAOYSA-N pentane-2,3-dione Chemical compound CCC(=O)C(C)=O TZMFJUDUGYTVRY-UHFFFAOYSA-N 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- IFBHRQDFSNCLOZ-RMPHRYRLSA-N 4-nitrophenyl beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C([N+]([O-])=O)C=C1 IFBHRQDFSNCLOZ-RMPHRYRLSA-N 0.000 description 1
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- WNBCMONIPIJTSB-BGNCJLHMSA-N Cichoriin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1)c1c(O)cc2c(OC(=O)C=C2)c1 WNBCMONIPIJTSB-BGNCJLHMSA-N 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 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 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 1
- 206010027304 Menopausal symptoms Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 1
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- CSCPPACGZOOCGX-UHFFFAOYSA-N acetone Substances CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 235000019631 acid taste sensations Nutrition 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 235000020244 animal milk Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 235000013527 bean curd Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 125000001721 carboxyacetyl group Chemical group 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- MEYVLGVRTYSQHI-UHFFFAOYSA-L cobalt(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Co+2].[O-]S([O-])(=O)=O MEYVLGVRTYSQHI-UHFFFAOYSA-L 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 235000015140 cultured milk Nutrition 0.000 description 1
- AJEHNBIPLQJTNU-UHFFFAOYSA-N cyanomethyl acetate Chemical compound CC(=O)OCC#N AJEHNBIPLQJTNU-UHFFFAOYSA-N 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940061607 dibasic sodium phosphate Drugs 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 1
- 235000005686 eating Nutrition 0.000 description 1
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 description 1
- 229940093496 esculin Drugs 0.000 description 1
- AWRMZKLXZLNBBK-UHFFFAOYSA-N esculin Natural products OC1OC(COc2cc3C=CC(=O)Oc3cc2O)C(O)C(O)C1O AWRMZKLXZLNBBK-UHFFFAOYSA-N 0.000 description 1
- 102000015694 estrogen receptors Human genes 0.000 description 1
- 108010038795 estrogen receptors Proteins 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000020981 healthy eating habits Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 235000006486 human diet Nutrition 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000001139 pH measurement Methods 0.000 description 1
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 description 1
- 239000003075 phytoestrogen Substances 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- MBDOYVRWFFCFHM-UHFFFAOYSA-N trans-2-hexenal Natural products CCCC=CC=O MBDOYVRWFFCFHM-UHFFFAOYSA-N 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 150000008495 β-glucosides Chemical class 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2445—Beta-glucosidase (3.2.1.21)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01021—Beta-glucosidase (3.2.1.21)
-
- 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
- A23V2400/137—Delbrueckii
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
本发明属于微生物技术领域,提供一种德氏乳杆菌及利用德氏乳杆菌发酵得到的豆乳饮料。公开了一株转化大豆异黄酮的德氏乳杆菌及其应用,具体为德氏乳杆菌(Lactobacillus delbrueckii)MT772183,保藏于中国典型培养物保藏中心(简称CCTCC),保藏编号为CCTCC NO:M 2020332,保藏日期为2020年07月20日。本发明的德氏乳杆菌MT772183具有较强的抗氧化能力、转化大豆异黄酮能力和产香去豆腥能力。本发明将德氏乳杆菌MT772183用于豆浆发酵,能去除豆乳的豆腥味,增加产品中的大豆异黄酮苷元含量,提高豆乳产品的抗氧化能力。
Description
技术领域
本发明涉及微生物技术领域,更具体地,涉及一种德氏乳杆菌及利用德氏乳杆菌发酵得 到的豆乳饮料。
背景技术
异黄酮是一类来自于植物根和种子的多酚化合物,其中大豆中的异黄酮含量最为丰富, 一般为1~2mg/g,因此大豆异黄酮是人类膳食中主要的异黄酮来源。大豆异黄酮的结构类似 于雌激素17-β-雌二醇(17-beta-estradiol),故大豆异黄酮又称为植物雌激素。大豆异黄酮具 有类似于雌激素的结构和功能,在多细胞动物体内可以通过与雌激素受体的相互作用调节雌 激素反应,从而起到预防更年期妇女心血管疾病、骨质疏松、乳腺癌和缓解更年期症状的作 用。大豆异黄酮还具有许多抗氧化活性,包括清除超氧自由基、抑制脂质过氧化、抑制与超 氧阴离子生成相关酶的合成等。除此之外,大豆异黄酮还具有降低胆固醇、预防Ⅱ型糖尿病、 预防冠心病、抗炎等多种生物学功能。
大豆异黄酮含有12种异构体,分为结合型糖苷和游离型苷元两类。其中,游离型苷元包 括染料木素(genistein)、大豆苷元(daidzein)和黄豆黄素苷元(glycitein)。结合型糖苷包 括染料木苷(genistin)、大豆苷(daidzin)、黄豆黄素(glycitin)及其相应的乙酰基和丙二 酰基衍生物。其中,染料木素、大豆苷元及其糖苷占天然大豆异黄酮总含量的95%以上,是 大豆异黄酮的主要组成成分。
大豆中约有97%~98%的异黄酮是以结合型糖苷的形式存在,而大豆异黄酮糖苷不易直 接被人体吸收,故天然大豆异黄酮的生物利用率十分有限,只有当大豆异黄酮糖苷被水解成 游离型苷元时才能被人体吸收、消化和利用。大多数结合型大豆异黄酮以β-葡萄糖苷的形式 存在。因此,人们普遍认为β-葡萄糖苷酶是水解大豆异黄酮糖苷的关键酶。β-葡萄糖苷酶能 够水解β-葡萄糖苷键,将染料木苷、大豆苷和黄豆黄素转化为对应的苷元形式。
乳酸菌作为人体肠道益生菌,具有多种生物学的功效,随着现代人们从健康饮食习惯向 绿色健康饮食方面的转变,益生菌非乳制品也越来越多地受到人们的喜爱。目前市面上的益 生菌乳制品大多为酸奶、乳饮料和乳酸菌饮料等。相较于动物乳所制成的乳制品,豆乳中的 蛋白质属于植物蛋白,含有丰富的不饱和脂肪酸,易被人体消化。但大豆中存在不易被人体 接受的豆腥味,人们普遍认为己醛、反-2-己烯醛、1-辛烯-3-醇、己醇、戊醇、乙酸、苯甲醛、 2,4-癸二烯醛这8种成分是豆腥味的风味物质。已有研究表明,用乳酸菌发酵豆浆,既可以降 低豆浆中部分豆腥味物质的含量,如己醛、2,4-癸二烯醛、1-辛烯-3-醇等,又能够产生乙酸、2,3- 丁二酮、2,3-戊二酮,3-羟基-2-丙酮等奶香味物质,从而构成酸豆乳的风味体系。若使用高产β- 葡萄糖苷酶的乳酸菌来发酵豆浆,既可以得到富含大豆异黄酮苷元的饮品,又能摄入一定的 益生菌,具有一定的应用价值,现有技术暂未发现能够高产β-葡萄糖苷酶的乳酸菌。
发明内容
本发明所要解决的技术问题是克服现有技术中存在的上述缺陷,首先提供一种德氏乳杆 菌。
本发明的第二个目的是提供上述德氏乳杆菌的应用。
本发明的目的通过以下技术方案实现:
本发明首先提供了一种德氏乳杆菌MT772183,其分类学命名为:德氏乳杆菌MT772183 (Lactobacillus delbrueckii MT772183),所述德氏乳杆菌于2020年7月20日保藏于中国典 型培养物保藏中心,保藏编号为CCTCC NO:M 2020332,保藏地址为:中国武汉大学。
本发明还提供所述的德氏乳杆菌在抗氧化方面的应用,所述的德氏乳杆菌MT772183的 羟基自由基清除率为24.02%,DPPH自由基清除率为16.99%,具备优异的抗氧化效果。
本发明还提供所述的德氏乳杆菌在发酵制备β-葡萄糖苷酶方面的应用。
优选地,通过发酵德氏乳杆菌的方式生产β-葡萄糖苷酶。
更优选地,利用德氏乳杆菌发酵生产β-葡萄糖苷酶的过程为:将德氏乳杆菌MT772183 接种于MRS液体培养基中,于37℃下培养24h,收集发酵液,即得β-葡萄糖苷酶。
更优选地,上述生产β-葡萄糖苷酶的过程中,也可以通过纯化发酵液得到高浓度的β-葡 萄糖苷酶。
本发明还提供所述的德氏乳杆菌在转化大豆异黄酮糖苷方面的应用。
优选地,利用德氏乳杆菌转化大豆异黄酮糖苷的过程为:将德氏乳杆菌MT772183经驯 化和厌氧培养后,接种于含有大豆异黄酮糖苷的液体培养基中,经37℃厌氧培养24h。
本发明中,德氏乳杆菌MT772183对大豆苷的转化率为96.17%,对染料木苷的转化率为 100.00%,对黄豆黄苷的转化率为81.33%,转化后的物质便于人体吸收利用,从而提高大豆 异黄酮在人体中的生物利用率。
本发明还提供一种豆乳饮料,由所述的德氏乳杆菌发酵得到。利用该德氏乳杆菌接种于 豆浆中,一方面,德氏乳杆菌可以将豆浆中的大豆异黄酮糖苷转化为游离型苷元,且转化率 高,便于人体吸收利用,另一方面,德氏乳杆菌在发酵过程中还生产抗氧化物质,因此得到 的豆乳饮料具有抗氧化能力;最后,德氏乳杆菌在发酵过程还能产生香气,并能够去除豆浆 中豆腥味。
与现有技术相比,本发明具有以下有益效果:
本发明提供了一株转化大豆异黄酮的德氏乳杆菌及其应用,具体为德氏乳杆菌(Lactobacillus delbrueckii)MT772183,保藏于中国典型培养物保藏中心(简称CCTCC),保藏编号为CCTCC NO:M 2020332,保藏日期为2020年07月20日。本发明的德氏乳杆菌MT772183具有较强的抗氧化能力和转化大豆异黄酮能力。本发明将德氏乳杆菌MT772183用于豆浆发酵,能去除豆乳的豆腥味,增加产品中的大豆异黄酮苷元含量,提高豆乳产品的抗 氧化能力;发酵能与其他三种功效协同进行,在发酵产酸凝固的同时达到良好的产香去豆腥 味效果,提高大豆苷元含量和抗氧化力。
附图说明
图1德氏乳杆菌MT772183菌落形态图;
图2德氏乳杆菌MT772183系统进化树;
图3MRS液体培养基提取物中大豆苷、大豆苷元含量示意图;
图4MT772183发酵物中大豆苷、大豆苷元含量示意图;
图5MRS液体培养基提取物中染料木苷、染料木素含量示意图;
图6MT772183发酵物中染料木苷、染料木素含量示意图;
图7豆浆发酵过程中pH变化趋势图;
图8豆浆发酵过程中酸度变化趋势图;
图9豆乳产品感官评定结果的描述性玫瑰图;
图10豆浆中大豆异黄酮含量示意图;
图11豆浆发酵48h后大豆异黄酮含量示意图。
具体实施方式
下面对本发明的具体实施方式作进一步说明。在此需要说明的是,对于这些实施方式的 说明用于帮助理解本发明,但并不构成对本发明的限定。此外,下面所描述的本发明各个实 施方式中所涉及的技术特征只要彼此之间未构成冲突就可以相互组合。
下述实验例中所使用的试验方法如无特殊说明,均为常规方法;所使用的材料、试剂等, 如无特殊说明,为可从商业途径得到的试剂和材料。
实施例1菌株的分离和鉴定
1、菌株的筛选
菌株分离基物酸浆水取自桂林腐乳传统生产工艺所用酸浆水,样品使用食品级PET瓶盛 装,于低温保存条件下带回实验室,置于4℃下保存。用无菌水将酸浆水进行梯度稀释,涂 布于含有2%碳酸钙的MRS培养基上,于37℃恒温培养箱中厌氧培养48h。
挑选产生溶钙圈的疑似乳酸菌菌落进行纯培养,编号为MT772183。重复纯化三次后, 挑取单个菌落进行革兰氏染色,观察到细胞为革兰氏阳性,长杆状,大多数成对生长,也有 单个出现。MT772183的菌落形态图见图1。
将MT772183单菌落接种于七叶苷显色培养基上,在37℃下培养24h后,菌落周围出现 明显棕黑色物质,表明MT772183能产生β-葡萄糖苷酶。
2、常规形态学及生理生化鉴定
进行革兰氏染色实验、接触酶实验、明胶液化实验、硝酸盐还原试验、石蕊牛奶实验、 甲基红实验、过氧化氢酶实验、葡萄糖产酸产气实验和碳水化合物发酵产酸试验(结果见表 1和表2)。参照《伯杰氏细菌学手册》及《乳酸细菌分类鉴定及实验方法》中乳杆菌属部分, 对MT772183进行初步鉴定,结果为德氏乳杆菌(Lactobacillus delbrueckii)。
表1 MT772183菌株的生理生化实验鉴定结果
表2 MT772183菌株的糖发酵实验结果
3、分子生物学鉴定
采用16S rDNA通用引物进行聚合酶链式反应(PCR)扩增。
正向引物为:27F:AGAGTTTGATCMTGGCTCAG;
反向引物为:1492R:GGTTACCTTGTTACGACTT。
PCR反应体系25μL:27F和1492R各1μL,Premix Taq DNA聚合酶12.5μL,灭菌双 蒸水9.5μL,37℃下恒温24h的培养液1μL。各反应物加入微量离心机离心10s后加入PCR 扩增仪。PCR反应程序:预变性95℃8min,变性95℃45s,复性55℃45s,延伸72℃2min, 25个循环,补充延伸72℃10min,保存12℃,5~10min。
PCR产物送美吉生物有限公司测序,与GenBank数据库中已知菌株的对应序列比较鉴定, 利用MEGA5.06构建系统进化树(见图2)。可知MT772183为德氏乳杆菌(Lactobacillus delbrueckii),其与Lactobacillus delbrueckii 6869的相似度为100%。
德氏乳杆菌MT772183保藏于中国典型培养物保藏中心(简称CCTCC),保藏地点为中 国武汉,保藏编号为CCTCC NO:M 2020332,保藏日期为2020年07月20日。
实施例2菌株β-葡萄糖苷酶活力的测定
测定德氏乳杆菌MT772183的发酵液与全细胞中β-葡萄糖苷酶的具体操作如下:
样品前处理:将德氏乳杆菌MT772183接种于MRS液体培养基中,于37℃下培养24h,取5mL发酵液于10mL离心管中,8000r/min离心20min,收集沉淀和上清液。菌体沉淀用 pH5.0磷酸二氢钾-磷酸氢二钠缓冲液洗涤两遍,离心去上清,沉淀溶于5mL pH 5.0的磷酸 二氢钾-磷酸氢二钠缓冲液中制成菌悬液,调节菌悬液的OD600值,使菌株的菌液密度接近109CFU/mL。
对硝基苯酚标准曲线绘制:准确称取对硝基苯酚139mg,用蒸馏水定容至1000mL。分 别吸取0.1mL、0.2mL、0.3mL、0.4mL、0.5mL、0.6mL、0.7mL、0.8mL、0.9mL溶液至 10mL容量瓶中,各加入2mL 1mol/L Na2CO3溶液,用蒸馏水定容至10mL,得浓度为0.01、 0.02、0.03、0.04、0.05、0.06、0.07、0.08、0.09mmol/L的稀释液。以1mol/L的Na2CO3溶 液作为空白对照,于400nm处测定吸光值(OD),以对硝基苯酚浓度为横坐标,最大吸收 峰处的吸光值为纵坐标,绘制标准曲线。
菌株的酶活力测定:在装有0.5mL菌悬液或发酵上清液的5mL离心管中,依次加入0.5 mL pH5.0磷酸二氢钾-磷酸氢二钠缓冲溶液和0.5mL 5mmol/L对硝基苯-β-葡萄糖苷溶液。 37℃水浴保温反应40min,立即加入2.0mL 1mol/L冷Na2CO3溶液终止反应。取反应后的溶液离心(8000r/min,15min),吸取上清液200μL于酶标孔板中,在400nm处测定其吸 光值。空白对照组:先用2mL 1.0mol/L的冷的Na2CO3使菌悬液或发酵上清液的酶失活,再 加入0.5mL pH 5.0磷酸二氢钾-磷酸氢二钠缓冲溶液和0.5mL 5mmol/L对硝基苯-β-葡萄糖苷溶液。样品的酶活计算如下:
Um—单位体积样品中β-葡萄糖苷酶活力(U);
X—将吸光值代入标准曲线后所得对硝基苯酚浓度(μmol/L);
V1—总反应液体积(L);
N—原酶液稀释倍数;
T—反应时间(min);
V2—样品体积(mL)。
实验测得德氏乳杆菌MT772183全细胞与发酵液的β-葡萄糖苷酶活力分别为3.08U/mL 和0.03U/mL。
实施例3菌株的抗氧化能力测定
1、无细胞提取物的制备
将德氏乳杆菌MT772183接种于MRS液体培养液中,于37℃培养24h,离心收集菌体,用pH7.4的PBS缓冲液洗涤3次后重悬于等量缓冲液中,调整细菌数量在1010CFU/ml左右, 冰浴超声波破碎细胞,4℃,12000g,离心30min。收集上清液,得到无细胞提取物。
2、清除羟基自由基的能力
在试管中加入2.5mmol/L的O-菲啰啉、pH7.4的PBS缓冲液、蒸馏水各1mL,充分混匀后加入1mL2.5mmol/L硫酸亚铁和1mL 0.1%H2O2,37℃水浴保温1h,在536nm处测 其吸光度记为A1;用1mL蒸馏水代替1mL H2O2的OD536值为A0;用1mL德氏乳杆菌 MT772183无细胞提取物代替1mL蒸馏水的OD536值为A2,平行测定三次。德氏乳杆菌 MT772183的羟基自由基清除率计算公式如下:
3、清除DPPH自由基的能力
在10ml离心管中加入1ml 0.2mmol/L DPPH自由基甲醇溶液和1ml德氏乳杆菌MT772183无细胞提取物,室温下避光反应30min。加入2ml的氯仿抽提,于4℃,3500r/min,离心10min后取0.1ml上清液,于517nm处测其吸光度,记为A1,平行测定三次。用蒸馏 水作为空白对照,其吸光度记为A0。德氏乳杆菌MT772183的DPPH自由基清除率计算公式 如下:
实验结果表明,德氏乳杆菌MT772183的羟基自由基清除率为24.02%,DPPH自由基清 除率为16.99%。
实施例4菌株对大豆异黄酮糖苷的转化能力
将菌株在MRS液体培养基中驯化1次后,将厌氧培养了5~6h的德氏乳杆菌MT772183 菌液以10%(v/v)的接种量分别接种于添加了100μM染料木苷和100μM大豆苷的MRS 液体培养基(添加了0.5g/L的L-半胱氨酸盐酸盐)中,37℃厌氧培养24小时,于-80℃下储 存,以加了糖苷且未接种的MRS液体培养基作为对照。
取5ml混匀后的培养液于25ml容量瓶中,加入80%的甲醇水溶液至接近刻度,混匀, 在超声波清洗器中提取20min,定容,8000r/min离心15min,收集上清液。取1ml滤液过0.45μm有机膜,-20℃储存用于测定。
高相液相色谱条件:
流动相为0.1%乙酸水溶液和0.1%乙酸乙腈溶液,按表3的方式进行梯度洗脱,柱温为 40℃,流速为1.0ml/min,进样量为20μL,检测波长为260nm。
表3梯度洗脱表
时间/min | 0.1%乙酸水溶液/ml | 0.1%乙酸乙腈溶液/ml |
0 | 90 | 10 |
12.5 | 70 | 30 |
17.5 | 60 | 40 |
18.5 | 0 | 100 |
21 | 0 | 100 |
22.5 | 90 | 10 |
26 | 90 | 10 |
由表4可知,德氏乳杆菌MT772183对大豆苷和染料木苷的转化率分别为100%和76%。 转化前后的大豆异黄酮HPLC图谱见图3至图6。
表4 MRS液体培养基中大豆异黄酮的含量
大豆苷转化率(%)=(转化前大豆苷含量-转化后大豆苷含量)/转化前大豆苷含量。
染料木苷转化率(%)=(转化前染料木苷含量-转化后染料木苷含量)/转化前染料木苷含量。
实施例5菌株的豆浆发酵特性
1、豆浆发酵剂的制备
将厌氧培养了5~6 h的德氏乳杆菌MT772183液体培养基在8000r/min,4℃下离心10min,用无菌水洗涤2次,将菌体重悬于等体积的无菌水中。以4%(v/v)的接种量接种于已灭菌的豆浆,37℃培养6~8 h,待豆浆凝固后,4℃冷藏,备用。
2、发酵豆浆
发酵豆浆的工艺流程如下:
大豆原料→室温浸泡12h→人工脱皮→磨浆(料水比1:5,80℃热水)→加热(80℃,15min)→添加1%(w/v)蔗糖→过滤→杀菌(121℃,15min)→冷却(50℃,10min)→ 冷却至37℃→以4%(w/v)的接种量接种→37℃发酵。
3、发酵豆乳的pH变化与酸度测定
分别测定豆浆发酵0h、2h、4h、6h、8h、10h、12h、24h的pH值和滴定酸度。pH测 定采用精密pH计测量。滴定酸度的测定方法参考GB-5009.239-2016酚酞指示法中的发酵乳 的酸度测定方法进行测定,具体方案为:取发酵豆乳10g(精确到0.001g)至150mL的锥形 瓶中,加入20mL新煮沸并冷却至室温的蒸馏水,混匀后加入2.0mL酚酞指示液,用0.1 mmol/L氢氧化钠标准溶液边滴定边摇晃,并不断向锥形瓶吹入氮气,至颜色与七水硫酸钴参 比溶液的颜色相似,且5s中内不消退,滴定应在45s内完成。空白试验以等体积的不含二 氧化碳的蒸馏水代替样品,读取消耗的氢氧化钠标准溶液的毫升数V0。发酵豆乳的酸度数值 以°T表示,按下式计算:
式中:X—试样的酸度,单位为度(°T)[以100g样品所消耗的0.1mol/L氢氧化钠毫升 数计,单位为毫升每100克(mL/100g)];
C1—氢氧化钠标准溶液的摩尔浓度,单位为摩尔每升(mol/L);
V1—滴定时所消耗氢氧化钠标准溶液的体积,单位为毫升(mL);
V0—空白实验所消耗氢氧化钠标准溶液的体积,单位为毫升(mL);
100—100g试样;
m—试样的质量,单位为克(g);
0.1—酸度理论定义氢氧化钠的摩尔浓度,单位为摩尔每升(mol/L)。
4、感官评定
待豆乳凝固后于4℃冷藏12h,参考GB/T 30885-2014《植物蛋白饮料豆奶和豆奶饮料》, 结合产品特点,制定感官评价方案,如表5所示。食品专业有感官评定经验的10人组成品评 小组进行评分,去掉每项中最高分和最低分各一个,结果取8人平均分。采用酸奶生产用商 业菌株、未接种的豆浆作对比。
表5发酵豆浆感官评价方案
由图7可知,发酵4h后,豆浆的pH值呈显著下降趋势,发酵8h后,豆浆pH值变化 平缓,继续发酵,pH值始终趋于4.70。由图8可知,豆浆酸度显著上升的趋势出现在发酵 6h后,且在后续发酵过程中呈递增趋势,发酵24h后,豆浆的酸度达67.31°T。由图9可知, 由德氏乳杆菌MT772183发酵而来的豆乳产品能起到良好的产香和去除豆腥味的效果。
实施例6豆浆发酵过程中大豆异黄酮的转化情况
用预先在豆浆中活化的菌株以4%(v/v)的接种量接种于装有30ml豆浆的50ml离心管 中,在37℃下发酵48h,于0、3、6、9、12、24、36、48h无菌取样,立即冰冷,测定大豆 异黄酮含量。以在相同实验条件下孵化的未接种豆浆为对照。
样品处理:取5ml混匀后的培养液于25ml容量瓶中,加入80%的甲醇水溶液至接近刻 度,混匀,在超声波清洗器中提取20min,定容,于8000r/min下离心15min,收集上清液,取1ml滤液过0.45μm有机膜。以在相同实验条件下孵化的未接种豆浆为对照。
高效液相色谱条件:流动相为0.1%乙酸水溶液和0.1%乙酸乙腈溶液,按表3的方式进行 梯度洗脱,柱温为40℃,流速为1.0ml/min,进样量为20μL,检测波长为260nm。
豆浆发酵前后色谱图如图10、图11所示。由表6、表7可知,豆浆发酵48h后,德氏 乳杆菌MT772183对大豆苷、染料木苷和黄豆黄素的转化率分别为96.17%、100.00%和81.33%。其中,在发酵6h后,即豆浆酸度开始显著增加时,德氏乳杆菌MT772183对大豆 苷、染料木苷和黄豆黄素的转化率分别为30.31%、26.75%和23.77%。而在发酵12h,豆浆 pH趋于稳定时,德氏乳杆菌MT772183对大豆苷、染料木苷和黄豆黄素的转化率分别为42.22%、100%和35.79%,且在后续的发酵过程中,德氏乳杆菌MT772183对大豆异黄酮的转化率持续增加,豆浆的酸度也在持续增加,但豆浆的酸度仍处于商业酸奶的酸度范围之内, 因此,延长发酵时间可提升异黄酮转化但不会影响产品酸感与风味。值得注意的是,在发酵12h后,豆浆中的总糖苷变化量为76.47μg/ml,显著高于已报道的产酸、产β-葡萄糖苷酶能力 强的菌株,如鼠李糖乳杆菌CRL981、植物乳杆菌KFRI144等。
表6豆浆发酵过程中大豆异黄酮的含量变化
表7德氏乳杆菌MT772183对大豆异黄酮的转化率变化
序列表
<110> 广西大学
<120> 一种德氏乳杆菌及利用德氏乳杆菌发酵得到的豆乳饮料
<130> ZM201351ZL
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213> Lactobacillus delbrueckii
<400> 1
agagtttgat cmtggctcag 20
<210> 2
<211> 19
<212> DNA
<213> Lactobacillus delbrueckii
<400> 2
ggttaccttg ttacgactt 19
<210> 3
<211> 748
<212> DNA
<213> Lactobacillus delbrueckii
<400> 3
gggtgatttg ttggatgcta gcggcggatg ggtgagtaac acgtgggcaa tctgccctaa 60
agactgggat accacttgga aacaggtgct aataccggat aacaacatga atcgcatgat 120
tcaagtttga aaggcggcgc aagctgtcac tttaggatga gcccgcggcg cattagctag 180
ttggtggggt aaaggcctac caaggcaatg atgcgtagcc gagttgagag actgatcggc 240
cacattggga ctgagacacg gcccaaactc ctacgggagg cagcagtagg gaatcttcca 300
caatggacgc aagtctgatg gagcaacgcc gcgtgagtga agaaggtctt cggatcgtaa 360
agctctgttg ttggtgaaga aggatagagg cagtaactgg tctttatttg acggtaatca 420
accagaaagt cacggctaac tacgtgccag cagccgcggt aatacgtagg tggcaagcgt 480
tgtccggatt tattgggcgt aaagcgagcg caggcggaat gataagtctg atgtgaaagc 540
ccacggctca accgtggaac tgcatcggaa actgtcattc ttgagtgcag aagaggagag 600
tggaactcca tgtgtagcgg tggaatgcgt agatatatgg aagaacacca gtggcgaagg 660
cggctctctg gtctgcaact gacgctgagg ctcgaaagca tgggtagcga acaggattag 720
ataccctggt agtccatgcc gtaaacga 748
Claims (6)
1.一种德氏乳杆菌,其特征在于,所述德氏乳杆菌于2020年07月20日保藏于中国典型培养物保藏中心,保藏编号为CCTCC NO:M 2020332,保藏地址为:中国武汉大学。
2.权利要求1所述的德氏乳杆菌在抗氧化方面的应用。
3.权利要求1所述的德氏乳杆菌在发酵制备β-葡萄糖苷酶方面的应用。
4.权利要求1所述的德氏乳杆菌在转化大豆异黄酮糖苷方面的应用。
5.根据权利要求4所述的应用,其特征在于,所述大豆异黄酮糖苷包括但不限于大豆苷、染料木苷、黄豆黄素。
6.一种豆乳饮料,其特征在于,由权利要求1所述的德氏乳杆菌发酵得到发酵豆乳,具有去除豆腥味的效果。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011205568.9A CN112574905B (zh) | 2020-11-02 | 2020-11-02 | 一种德氏乳杆菌及利用德氏乳杆菌发酵得到的豆乳饮料 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011205568.9A CN112574905B (zh) | 2020-11-02 | 2020-11-02 | 一种德氏乳杆菌及利用德氏乳杆菌发酵得到的豆乳饮料 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112574905A true CN112574905A (zh) | 2021-03-30 |
CN112574905B CN112574905B (zh) | 2022-03-22 |
Family
ID=75120058
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202011205568.9A Active CN112574905B (zh) | 2020-11-02 | 2020-11-02 | 一种德氏乳杆菌及利用德氏乳杆菌发酵得到的豆乳饮料 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112574905B (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113558193A (zh) * | 2021-06-24 | 2021-10-29 | 苏州大学 | 一种富含大豆异黄酮苷元的发酵饮料的制备方法 |
CN114854623A (zh) * | 2022-03-15 | 2022-08-05 | 泸州老窖股份有限公司 | 德氏乳杆菌保加利亚亚种、含有该菌的菌剂及应用 |
-
2020
- 2020-11-02 CN CN202011205568.9A patent/CN112574905B/zh active Active
Non-Patent Citations (4)
Title |
---|
YEON-BAE CHOI等: "Hydrolysis of soybean isoflavone glucosides by lactic acid bacteria", 《BIOTECHNOLOGY LETTERS》 * |
YOUNG-HEE PYO等: "Effect of Lactic Acid Fermentation on Enrichment of Antioxidant Properties and Bioactive Isoflavones in Soybean", 《JOURNAL OF FOOD SCIENCE》 * |
于国萍 等: "乳酸菌发酵法水解大豆异黄酮", 《食品科学》 * |
白宝兰,刘妍,李想主编: "《食品工艺学》", 30 June 2018, 北京工业大学出版社 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113558193A (zh) * | 2021-06-24 | 2021-10-29 | 苏州大学 | 一种富含大豆异黄酮苷元的发酵饮料的制备方法 |
CN114854623A (zh) * | 2022-03-15 | 2022-08-05 | 泸州老窖股份有限公司 | 德氏乳杆菌保加利亚亚种、含有该菌的菌剂及应用 |
CN114854623B (zh) * | 2022-03-15 | 2023-09-29 | 泸州老窖股份有限公司 | 德氏乳杆菌保加利亚亚种、含有该菌的菌剂及应用 |
Also Published As
Publication number | Publication date |
---|---|
CN112574905B (zh) | 2022-03-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Wei et al. | Using of Lactobacillus and Bifidobacterium to product the isoflavone aglycones in fermented soymilk | |
CN1322140C (zh) | 含异黄酮的组合物 | |
Chun et al. | Conversion of isoflavone glucosides to aglycones in soymilk by fermentation with lactic acid bacteria | |
Marazza et al. | Aglycone production by Lactobacillus rhamnosus CRL981 during soymilk fermentation | |
Chun et al. | Enrichment of isoflavone aglycones in soymilk by fermentation with single and mixed cultures of Streptococcus infantarius 12 and Weissella sp. 4 | |
CN112574905B (zh) | 一种德氏乳杆菌及利用德氏乳杆菌发酵得到的豆乳饮料 | |
CN100364454C (zh) | 一种生产风干牛肉的方法 | |
CN109402000A (zh) | 一株产β-葡萄糖苷酶乳酸菌及其筛选方法和富含活性黄酮苷元酸奶的制备方法 | |
Otieno et al. | Endogenous β‐glucosidase and β‐galactosidase activities from selected probiotic micro‐organisms and their role in isoflavone biotransformation in soymilk | |
CN111197018B (zh) | 一种酸鱼乳杆菌、用其发酵豆乳的方法及制备出的发酵豆乳与应用 | |
KR20110056419A (ko) | 에쿠올 생산능이 유지된 에쿠올 생산 미생물을 포함하는 발효 제품 및 그의 제조 방법 | |
CN107006606A (zh) | 一种富含功能因子的风味酸奶及其制备方法 | |
CN107674852A (zh) | 一种高产丁二酮的植物乳杆菌及其应用 | |
CN102199565B (zh) | 一种屎肠球菌及其产生雌马酚的方法与应用 | |
Chun et al. | Hydrolysis of isoflavone glucosides in soymilk fermented with single or mixed cultures of Lactobacillus paraplantarum KM, Weissella sp. 33, and Enterococcus faecium 35 isolated from humans | |
CN117987298A (zh) | 一种植物乳植杆菌及采用该菌株制备芦笋发酵饮料的方法 | |
BORDIGNON et al. | Hydrolysis of isoflavones and consumption of oligosaccharides during lactic acid fermentation of soybean milk | |
CN116083312B (zh) | 一株高产β-葡萄糖苷酶的乳酸菌及其在发酵食品中的应用 | |
Chen et al. | Effect of oligosaccharides and isoflavones aglycones in defatted soy meal fermented by Lactobacillus paracasei and Bifidobacterium longum | |
KR101673055B1 (ko) | 신규한 스타터 균주 및 이를 이용한 두부를 포함하는 기능성 발효식품의 제조방법 | |
KR102530797B1 (ko) | 기능성 청국장 제조용 바실러스 리체니포미스 idck30와 바실러스 서브틸리스 idck40의 복합종균 조성물 및 이를 이용한 산양삼 청국장 제조방법 | |
KR101384580B1 (ko) | 스트렙토코커스속 유산균 스타터를 이용한 두유발효유 및그 제조방법 | |
Prasad et al. | Conversion of isoflavone glycoside to aglycones in soy protein isolate (SPI) using crude enzyme extracted from Bifidobacterium animalis Bb12 and Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842 | |
US6245536B1 (en) | Process for preparing isoflavone aglucone using Rhodotorula glutinis having an ability to produce isoflavone aglucone | |
CN104673722B (zh) | 耐氧夏普氏菌及其在二氢大豆异黄酮有氧合成中的应用 |
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 | ||
GR01 | Patent grant | ||
GR01 | Patent grant |