CN112661869A - 杂多糖 - Google Patents
杂多糖 Download PDFInfo
- Publication number
- CN112661869A CN112661869A CN202011108623.2A CN202011108623A CN112661869A CN 112661869 A CN112661869 A CN 112661869A CN 202011108623 A CN202011108623 A CN 202011108623A CN 112661869 A CN112661869 A CN 112661869A
- Authority
- CN
- China
- Prior art keywords
- heteropolysaccharide
- mol
- galactose
- glucose
- rhamnose
- 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.)
- Withdrawn
Links
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 34
- 239000008103 glucose Substances 0.000 claims abstract description 31
- 229930182830 galactose Natural products 0.000 claims abstract description 30
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims abstract description 29
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 claims abstract description 29
- 241000894006 Bacteria Species 0.000 claims abstract description 28
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 claims abstract description 28
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 claims abstract description 27
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 claims abstract description 27
- 150000002772 monosaccharides Chemical class 0.000 claims abstract description 24
- 235000021001 fermented dairy product Nutrition 0.000 claims abstract description 17
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 claims abstract description 14
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims description 14
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 12
- 238000004519 manufacturing process Methods 0.000 claims description 10
- 235000013618 yogurt Nutrition 0.000 claims description 10
- 241000194020 Streptococcus thermophilus Species 0.000 claims description 7
- 239000004310 lactic acid Substances 0.000 claims description 6
- 235000014655 lactic acid Nutrition 0.000 claims description 6
- 235000013351 cheese Nutrition 0.000 claims description 5
- 241000194017 Streptococcus Species 0.000 claims description 4
- 230000002538 fungal effect Effects 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims 1
- 229920002444 Exopolysaccharide Polymers 0.000 description 29
- 239000000203 mixture Substances 0.000 description 17
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- 235000013305 food Nutrition 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 7
- 235000013365 dairy product Nutrition 0.000 description 7
- 244000005700 microbiome Species 0.000 description 7
- 235000000346 sugar Nutrition 0.000 description 7
- 150000004676 glycans Chemical class 0.000 description 6
- 229920001282 polysaccharide Polymers 0.000 description 6
- 239000005017 polysaccharide Substances 0.000 description 6
- 235000015140 cultured milk Nutrition 0.000 description 5
- 238000000502 dialysis Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 238000001223 reverse osmosis Methods 0.000 description 5
- 238000001542 size-exclusion chromatography Methods 0.000 description 5
- 238000005903 acid hydrolysis reaction Methods 0.000 description 4
- 235000014048 cultured milk product Nutrition 0.000 description 4
- 235000013336 milk Nutrition 0.000 description 4
- 239000008267 milk Substances 0.000 description 4
- 210000004080 milk Anatomy 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- HNSDLXPSAYFUHK-UHFFFAOYSA-N 1,4-bis(2-ethylhexyl) sulfosuccinate Chemical compound CCCCC(CC)COC(=O)CC(S(O)(=O)=O)C(=O)OCC(CC)CCCC HNSDLXPSAYFUHK-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000003643 water by type Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 2
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 2
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 2
- 229920001222 biopolymer Polymers 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 235000019219 chocolate Nutrition 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 235000004213 low-fat Nutrition 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 description 1
- MSWZFWKMSRAUBD-GASJEMHNSA-N 2-amino-2-deoxy-D-galactopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O MSWZFWKMSRAUBD-GASJEMHNSA-N 0.000 description 1
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- GJCOSYZMQJWQCA-UHFFFAOYSA-N 9H-xanthene Chemical compound C1=CC=C2CC3=CC=CC=C3OC2=C1 GJCOSYZMQJWQCA-UHFFFAOYSA-N 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- AEMOLEFTQBMNLQ-YMDCURPLSA-N D-galactopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-YMDCURPLSA-N 0.000 description 1
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 241001600129 Delftia Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229920002148 Gellan gum Polymers 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 229920000869 Homopolysaccharide Polymers 0.000 description 1
- 235000008694 Humulus lupulus Nutrition 0.000 description 1
- 241000186673 Lactobacillus delbrueckii Species 0.000 description 1
- 241000192130 Leuconostoc mesenteroides Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- OVRNDRQMDRJTHS-RTRLPJTCSA-N N-acetyl-D-glucosamine Chemical compound CC(=O)N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-RTRLPJTCSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 241000736110 Sphingomonas paucimobilis Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 241000589636 Xanthomonas campestris Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- WQZGKKKJIJFFOK-UHFFFAOYSA-N alpha-D-glucopyranose Natural products OCC1OC(O)C(O)C(O)C1O WQZGKKKJIJFFOK-UHFFFAOYSA-N 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
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- 235000003599 food sweetener Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
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- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000000216 gellan gum Substances 0.000 description 1
- 235000010492 gellan gum Nutrition 0.000 description 1
- 239000003349 gelling agent Substances 0.000 description 1
- 235000021474 generally recognized As safe (food) Nutrition 0.000 description 1
- 235000021473 generally recognized as safe (food ingredients) Nutrition 0.000 description 1
- 229960002442 glucosamine Drugs 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
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- 238000002347 injection Methods 0.000 description 1
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- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
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- 235000020161 semi-skimmed milk Nutrition 0.000 description 1
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- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
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- 239000008399 tap water Substances 0.000 description 1
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- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 1
- 230000008719 thickening Effects 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 238000009849 vacuum degassing Methods 0.000 description 1
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- 229920001285 xanthan gum Polymers 0.000 description 1
- 235000008924 yoghurt drink Nutrition 0.000 description 1
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
-
- 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
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/02—Making cheese curd
- A23C19/032—Making cheese curd characterised by the use of specific microorganisms, or enzymes of microbial origin
- A23C19/0323—Making cheese curd characterised by the use of specific microorganisms, or enzymes of microbial origin using only lactic acid bacteria, e.g. Pediococcus and Leuconostoc species; Bifidobacteria; Microbial starters in general
-
- 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
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/02—Making cheese curd
- A23C19/05—Treating milk before coagulation; Separating whey from curd
- A23C19/054—Treating milk before coagulation; Separating whey from curd using additives other than acidifying agents, NaCl, CaCl2, dairy products, proteins, fats, enzymes or microorganisms
-
- 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
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
- A23C19/09—Other cheese preparations; Mixtures of cheese with other foodstuffs
-
- 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
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
-
- 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
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1238—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt using specific L. bulgaricus or S. thermophilus microorganisms; using entrapped or encapsulated yoghurt bacteria; Physical or chemical treatment of L. bulgaricus or S. thermophilus cultures; Fermentation only with L. bulgaricus or only with S. thermophilus
-
- 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
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
-
- 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
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/137—Thickening substances
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L5/00—Compositions of polysaccharides or of their derivatives not provided for in groups C08L1/00 or C08L3/00
-
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Abstract
本发明涉及杂多糖及其生产方法,所述杂多糖的特征在于:它基本上由单糖葡萄糖、半乳糖、鼠李糖和N‑乙酰半乳糖胺构成。本发明还涉及能够产生所述杂多糖的细菌以及所述杂多糖和所述细菌用于改善发酵乳制品的质地的用途。
Description
本申请是申请号为201480060300.7名称为“杂多糖”的中国专利申请 的分案申请,原申请是申请日为2014年11月3日的国际申请 PCT/EP2014/073570的中国国家阶段申请。
技术领域
本发明涉及杂多糖、其通过微生物的生产及其在乳制品行业中用于制 造发酵乳制品的应用。
背景技术
来自植物、动物和微生物来源的聚合物在食品配方中起着重要的作 用。食品聚合物是长链的高分子质量分子,其溶解或分散在水中从而提供 质地化(texturizing)特性。食品行业所使用的大多数生物聚合物是来自作 物植物的多糖(例如,淀粉)或者来自海藻类的多糖(例如,卡拉胶)以 及动物蛋白如酪蛋白酸盐和明胶。植物碳水化合物在食品中的功能性质由 相当微妙的结构特征决定。对于工业实践,大部分被化学改性。
替代性的生物增稠剂是微生物表多糖。这些聚合物可被组装成与细胞 表面紧密相连的荚膜多糖(CPS),或者它们可被释放到生长介质中 (即,“丝状(ropy)”多糖)。术语表多糖(EPS)可用来描述任一种 类型的胞外多糖(Broadbent J.R.等人,2003,JournalDairy Science,86,407- 423,“Biochemistry,genetics and applications ofexopolysaccharide production in Streptococcus thermophilus:a review”)。
EPS广泛存在于细菌和微藻中并较少存在于酵母和真菌中。工业上重 要的微生物EPS的实例是来自Leuconostoc mesenteroides的右旋糖酐、来 自Xanthomonascampestris的黄原胶和来自Sphingomonas paucimobilis的结 兰胶家族的EPS。这些EPS仅代表目前生物聚合物市场的一小部分。限制 微生物EPS的使用的因素是其经济生产(这需要对其生物合成的充分了解 和适应性生物工艺技术)、其高回收成本以及其中大多数是非食品细菌来 源。对于EPS的食品用途,能够足够大量地产生EPS的公认安全 (GRAS)的食品级微生物、尤其是乳酸菌(LAB)的菌株是令人关注的 替代物。此外,这些微生物可被用于在原位产生EPS,对于发酵乳制品尤 其如此,以改善它们的流变性、质地和主体,以及口感。
来自LAB的EPS可被细分成两组,即同多糖(HoPS)和杂多糖 (HePS)。HoPS由一种类型的组成单糖(d-吡喃葡萄糖或d-呋喃果糖)构成 (Monsan,P.等人,2001,InternationalDairy Journal,11,673–683, Homopolysaccharides from lactic acid bacteria.)。HePS由重复子单元的骨 架构成,所述重复子单元是支化的(在C2、C3、C4或C6位置)或未支化的并且由3-8个单糖、单糖衍生物或经取代的单糖组成。最近的综述请 参见Vaningelgem等人(2004)applied and Environmental Microbiology 70(2), 900-912“Biodiversityof Exopolysaccharides Produced by Streptococcus thermophilus Strains IsReflected in Their Production and Their Molecular and FunctionalCharacteristics”和De Vuyst等人(2001)International Dairy Journal, 11,687-707.“Recent developments in the biosynthesis and applications ofheteropolysaccharides from lactic acid bacteria”。
de Vuyst等人(2001)中的表1总结了34种不同乳酸菌的HePS。重复 单元包含以下糖类:葡萄糖(Glc)、GlcNAc(N-乙酰葡糖胺)、半乳糖、 (Gal)、GalNAc(N-乙酰半乳糖胺)、鼠李糖(Rha)和海藻糖(Fuc)。 Vaningelgem等人中的表4总结了25种不同Streptococcusthermophilus菌 株的HePS。而de Vuyst(2001)基于重复单元中单糖的数目对不同EPS进行分类,从三-糖开始直至八-糖,Vaningelgem等人根据它们的定性单糖 组成定义了组I-组VI。例如,组I由半乳糖和葡萄糖构成,而组V由半乳 糖、葡萄糖、鼠李糖和N-乙酰半乳糖胺构成。Ruas-Madiedo等人,2002, International Dairy Journal,12,163-171,Anoverview of the functionality of exopolysaccharides produced by lactic acidbacteria的表2中总结了HePS的 重复单元的额外结构。
众所周知,EPS的功能受分子的3D结构(即,所谓的流体力学半径- Rh)的影响。决定3D结构的两个主要参数是分子质量(MW,以道尔顿 计)和EPS的重复单元,其中EPS的重复单元由聚合物中分支与连接的 类型、单糖组成决定。
来自LAB的EPS在一些发酵乳制品的生产中具有技术重要性。使用 含有形成EPS的S.thermophilus和Lactobacillus delbrueckii subsp. bulgaricus的“丝状”起子(starter)是生产酸乳中的常用做法以改善酸乳 的质地、避免酸乳脱水收缩和提高酸乳的粘度。此外,产生EPS的细菌被 用来改善奶酪、尤其是低脂奶酪的产率、质地和功能(参见Broadbent J.R. 等人,2003,Journal Dairy Science,86,407-423,“Biochemistry,genetics and applications of exopolysaccharide production in Streptococcusthermophilus:a review”)。
我们现在已经出乎意料地发现了具有卓越质地化特性的新型杂多糖。 杂多糖的分子重量和糖组成的组合以前尚未被报道过。
发明内容
在第一方面,本发明提供杂多糖,所述杂多糖基本上由单糖葡萄糖、 半乳糖、鼠李糖和N-乙酰半乳糖胺构成并且具有100kDa-10000kDa的分子 重量。基本上在本文中被定义为:基于杂多糖的干物质,至少90%、优选 地至少95%、更优选地至少98%的杂多糖由单糖葡萄糖、半乳糖、鼠李糖 和N-乙酰半乳糖胺构成。在一个优选的实施方式中,杂多糖由30-50摩尔% 的葡萄糖、10-30摩尔%的半乳糖、10-30摩尔%的鼠李糖和10-30摩尔%的 N-乙酰半乳糖胺构成,且其中葡萄糖、半乳糖、鼠李糖和N-乙酰半乳糖胺 的总摩尔%为100%。更优选地,杂多糖由35-45摩尔%的葡萄糖、15-25摩 尔%的半乳糖、15-25摩尔%的鼠李糖和15-25摩尔%的N-乙酰半乳糖胺构 成,优选地其中葡萄糖、半乳糖、鼠李糖和N-乙酰半乳糖胺的总摩尔%为 100%。在另一个实施方式中,杂多糖的特征在于:葡萄糖的摩尔%与半乳 糖的摩尔%的比率在≥1.0和≤2.0之间。最优选地,杂多糖由40摩尔%的葡萄 糖、20摩尔%的半乳糖、20摩尔%的鼠李糖和20摩尔%的N-乙酰半乳糖胺 构成。
根据本发明的杂多糖的组成可以以几种方式被表示。组成可以被表示 成各单糖的重量百分比或者被表示成各单糖的摩尔百分比。或者,摩尔百 分比可以被表示成各单糖的摩尔比(整数)。例如,上文所述的最优选实 施方式(其由40摩尔%的葡萄糖、20摩尔%的半乳糖、20摩尔%的鼠李糖 和20摩尔%的N-乙酰半乳糖胺构成)则被表示成:
葡萄糖:半乳糖:鼠李糖:N-乙酰半乳糖胺=2:1:1:1。
本发明的杂多糖可以由基本重复单元构成。重复单元可以具有与杂多 糖的组成相同的组成。优选地,本发明的杂多糖的重复单元是由葡萄糖、 半乳糖、鼠李糖和N-乙酰半乳糖胺构成的五糖。五糖在本文中被定义为由 5个单糖构成的糖。本发明的五糖优选地具有以下组成:
葡萄糖:半乳糖:鼠李糖:N-乙酰半乳糖胺=2:1:1:1。
本发明的杂多糖具有100kDa-10000kDa的分子重量。更优选地200 kDa-10000kDa的分子重量,优选地300kDa-10000kDa的分子重量,优选 地400kDa-10000kDa的分子重量,优选地450kDa-10000kDa的分子重量, 优选地500kDa-10000kDa的分子重量,更优选地200kDa-8000kDa的分子 重量,优选地300kDa-6000kDa的分子重量,优选地400kDa-4000kDa的分 子重量,优选地450kDa-2000kDa的分子重量,最优选地500kDa-1500kDa 的分子重量。本发明的杂多糖的分子重量在本文中被定义为平均分子重 量。杂多糖的分子重量将具有在平均分子重量左右的分子重量分布。平均 分子重量可以由技术人员通过本领域已知的方法(例如尺寸排阻色谱法) 来测定。
本发明的杂多糖可以具有上文所述的任何糖组成与上述所述的任何分 子重量范围的组合。一个高度优选的本发明的杂多糖由35-45摩尔%的葡萄 糖、15-25摩尔%的半乳糖、15-25摩尔%的鼠李糖和15-25摩尔%的N-乙酰 半乳糖胺构成,其中葡萄糖、半乳糖、鼠李糖和N-乙酰半乳糖胺的总摩 尔%为100%,并且具有500kDa-1500kDa的分子重量。另一高度优选的本 发明的杂多糖具有以下糖组成:
葡萄糖:半乳糖:鼠李糖:N-乙酰半乳糖胺=2:1:1:1, 并且具有500kDa-1500kDa的分子重量。
本发明的杂多糖的优点为:相较于虽然由相同的单糖葡萄糖、半乳 糖、鼠李糖和N-乙酰半乳糖胺构成但具有比100kDa低得多的分子质量的 杂多糖,其在发酵乳产品中具有卓越的增稠性质并且提供长纹理(long texture)(参见实施例)。
在第二方面,本发明提供能够产生本发明的杂多糖的的细菌。优选 地,所述细菌属于Streptococcus种。更优选地,所述细菌选自由以下组成 的组
·Streptococcus salivarius thermophilus NGB-22D,于2012年2月28日在 乌特勒支的Centraalbureau voor Schimmelcultures(真菌生物多样性中心)保 藏为CBS132067
·Streptococcus salivarius thermophilus DS65008,于2013年6月18日在 乌特勒支的Centraalbureau voor Schimmelcultures(真菌生物多样性中心)保 藏为CBS135685。
本发明的细菌优选地产生上文所限定的杂多糖,即基本上由单糖葡萄 糖、半乳糖、鼠李糖和N-乙酰半乳糖胺构成的杂多糖。所述细菌能够产生 在本发明的第一方面中限定的具有优选组成的杂多糖。本发明的细菌还能 够产生在本发明的第一方面中限定的由基本重复单元构成的杂多糖。
在第三方面,本发明提供生产发酵乳制品的方法,所述方法包括:添 加在本发明的第一方面中限定的本发明的杂多糖。添加本发明的杂多糖的 效果是:相较于不添加杂多糖或者相较于添加具有不同组成的杂多糖,获 得了高度粘稠、可拉伸和光滑质地的发酵乳制品。
在另一实施方式中,本发明提供生产发酵乳制品的方法,所述方法包 括:添加在本发明的第二方面中限定的能够产生杂多糖的本发明的细菌。 在该实施方式中,在用于生产发酵乳制品的方法中,所述细菌在原位产生 杂多糖,相较于不添加本发明的细菌或者相较于添加能够产生具有不同组 成的多糖的细菌,这导致发酵乳制品的质地和/或粘度得到改善。
发酵制品可以是期望质地和/或粘度得到改善的任何发酵乳制品。发酵 乳制品可以选自,但不限于,发酵制品例如酸乳、饮用酸乳、(低脂)奶 酪、开菲尔(kefir)、酪乳、酸乳油或大豆酸乳等。这类食品还可包含用 于制备甜点的常见成分,例如水果、巧克力片或谷物,以及甜化产品或液 体巧克力。食品还可包含常见的食物成分例如乳化剂、胶凝剂、稳定剂、 甜味剂等。本领域技术人员知道如何使用来自本发明的(发酵)食物来制 备食品。
在第四方面,本发明提供生产在本发明的第一方面中限定的本发明的 杂多糖的方法。优选地,在生产本发明的杂多糖的方法中,在有益于产生 本发明的杂多糖的条件下,在培养基中培养本发明的第二方面中限定的本 发明的细菌。可以使用本领域已知的技术从发酵液中收集本发明的杂多 糖,所述发酵液包含细菌的生物质、本发明的杂多糖和其它组分。
在第五方面,本发明提供在本发明的第一方面中限定的本发明的杂多 糖用于改善发酵乳制品的质地和/或粘度的用途。优选地,发酵乳制品是酸 乳或奶酪。
在第六方面,本发明提供在本发明的第二方面中限定的本发明的细菌 用于改善发酵乳制品的质地和/或粘度的用途。
具体实施方式
材料和方法
菌株
**Streptococcus thermophilus ST144获自乳制品微生物保藏中心(CultureCollection of Dairy Microorganisms)(Milcom),Sobeslavska 841,39001Tabor,捷克共和国
***菌株于指定日期(保藏日期)被保藏在Centraalbureau voorSchimmelcultures(真菌生物多样 性中心),Uppsalalaan 8,3508AD乌特勒支,荷兰,并收到的相应的保藏号
制备发酵乳制品
向商业UHT半脱脂乳(Campina,荷兰)补充2%(w/w)的脱脂乳粉末。 如下对200ml乳的等分试样进行巴氏灭菌:在水浴中在90℃下加热乳15分 钟,随后在85℃下加热30分钟。巴氏灭菌之后,利用冰水将乳迅速冷却至 4℃。用106cfu/ml(菌落形成单位/ml)各种细菌培养物接种巴氏灭菌乳并在 42℃孵育直至pH达到4.6。使用CINAC设备(Ysebaert,法国)连续监测培养 物的pH。同时,为了测量Brookfield粘度,在杯子(体积为150ml,直径为5.5cm)中以相同的方式发酵125ml乳。
在冰水中冷却如此得到的发酵乳并贮存在4℃直至进一步分析。
从发酵乳中分离EPS
在50mL管中在55℃下加热40mL发酵乳1小时以释放EPS。为了沉淀 蛋白质,加入2.67mL 60%(w/v)的TCA并在20-25℃之间的温度下轻轻摇 动溶液1.5小时。在4℃的温度下以6300g离心30分钟之后,将上清液定量 转移到100mL瓶中并利用5N的氢氧化钠(NaOH)将pH调节至4.0。将上清液 定量转移到30cm的透析管(4个MWCO 12 000-14 000的SpectraPor膜, Spectrum Laboratories Inc)中并相对于自来水透析约16小时(过夜),随后 利用反渗透(RO)水透析2小时。使用前,在2%的碳酸钠(Na2CO3)溶液中煮 沸透析管15分钟,利用RO水洗涤两次,然后利用RO水再次煮沸15分钟。 将透析管贮存在RO水中直至使用。然后将透析液定量转移到塑料杯中(利 用5mL RO水洗涤透析管两次)并在两天期间冻干。
SEC-MALLS分析
尺寸排阻色谱(SEC)系统由串联的TSK凝胶PWXL保护柱(6.0mm×4.0cm)、TSK凝胶G6000 PWXL分析柱(7.8mm×30cm,13.0μm)和 TSK凝胶G5000 PWXL分析柱(7.8mm×30cm,10μm)(TosoHaas)组成 并利用温度控制模件(Waters)恒温控制在35℃。
对洗脱液进行在线真空脱气(1200系列,Agilent Technologies)并以0.5 mL/分钟的流量泵送(1200系列,Agilent Technologies)。将样品置于10℃ 的温控样品架中并在柱(模型231Bio,Gilson)上注射200μL。
于32°和144°之间的15个角度在632.8nm下测量光散射(DAWN DSP-F, WyattTechnologies)。在280nm下测量UV吸收(CD-1595,Jasco)以检测蛋白 质。
使用ASTRA V软件(5.3.2.22版,Wyatt Technologies)进行数据收集和处 理。为了归一化,使用参照物牛血清白蛋白(BSA)(M.W.=67000Da)。洗 脱液为100mM的NaNO3+0.02%NaN3。使用前,通过0.20μm的滤器并在 线通过0.025μm的滤器过滤洗脱液,其中0.025μm的滤器位于泵和注射器 之间。注射前,将冻干的EPS样品以25mg/mL的浓度溶解在洗脱液中并通 过0.20μm的滤器过滤。
酸水解EPS后分析单糖
酸水解多糖–在利用尺寸排阻色谱(SEC)分析期间,收集到了多糖峰(在 29-31分钟之间:2分钟×0.5mL/分钟=1mL)。加入TFA储液(1mL;4M) 并在氮气(N2)中利用2M三氟乙酸(TFA)在120℃下酸水解收集到的多糖溶 液75分钟。水解之后,于真空中在室温下过夜干燥溶液,然后将其溶解在 水(100μL)中并通过HPAEC-PAD分析。
HPAEC-PAD分析–在金电极上的高效阴离子交换色谱与脉冲安培检测法 (HPAEC-PAD)被用于单糖鼠李糖、半乳糖胺、阿拉伯糖、葡萄糖胺、 半乳糖、葡萄糖、甘露糖、木糖、半乳糖醛酸和葡糖醛酸的定量分析。利 用带有氦脱气单元的600E系统控制器泵(Waters)和模型400EC检测器 (EG&G),使用VT-03流动池(Antec Leyden)和Dionex的技术说明21中所述 的关于波形B的设置进行分析。利用717自动进样器(Waters),将20μL样品 注射在恒温控制在30℃的250×4mm(10-32)的Carbopac PA-1柱(Dionex) 上。以1.0mL/分钟的流速洗脱单糖。利用16mM的氢氧化钠等度洗脱单 糖,随后洗脱酸单糖:从20分钟开始在20分钟内以线性梯度达到200mM 氢氧化钠+500mM乙酸钠。
利用Chromeleon软件6.80版(Dionex)进行数据分析。利用经受酸水解条 件的单糖(Sigma-Aldrich)标准溶液(40-250mg/L之间)进行定量分析。
质地分析
通过用长塑料勺手动搅动杯子中的酸乳来定性评估酸乳的质地。通过 搅动期间观察到的阻力来评价样品的稠度和胶黏性。通过使一勺酸乳从勺 子流动来评价长(long)质地和短(short)质地。短酸乳将从勺子落下成 为一团,而长酸乳将从勺子流动并形成线(thread)。
实施例
实施例1
由S.thermophilus菌株产生的EPS的特征
如材料和方法中所述,利用不同的S.thermophilus菌株生产发酵乳并从 其中分离EPS。表1总结了发酵乳的质地和稠度以及EPS的糖组成及其分子 重量。
菌株BLF5-1充当对照,因为它提供易碎的粒状质地(“短”)。菌株 STV88是现有技术菌株,已知其相较于对照菌株BLF5-1提供平均质地化的 可拉伸的和光滑的(“长”)发酵品。菌株ST144充当对照,因为它具有 与本发明的菌株相同的糖组成但低得多的分子重量。该菌株也提供短质 地。
菌株NGB-22D和DS65008是代表本发明的菌株,其产生高度粘稠、可 拉伸和光滑(“长”)质地的发酵乳制品。
EPS的分析显示:菌株NGB-22D和DS65008产生这样的EPS,其相较于 比较菌株的EPS具有独特的糖组成和分子重量的组合
表1-S.thermophilus酸乳菌株经分离的EPS的糖组成
Glc=葡萄糖;Gal=半乳糖;Rha=鼠李糖;GalNAc=N-乙酰半乳糖胺。其它单糖未被包括 在内,因为它们的值低于检测限。ND=未检出。
*摘自Vaningelgem(2004)–表3和表4
Claims (14)
1.杂多糖,其特征在于:其基本上由单糖葡萄糖和半乳糖和鼠李糖和N-乙酰半乳糖胺构成,并且所述杂多糖具有100kDa-10000kDa的分子重量。
2.根据权利要求1所述的杂多糖,其特征在于:其由30-50摩尔%的葡萄糖、10-30摩尔%的半乳糖、10-30摩尔%的鼠李糖和10-30摩尔%的N-乙酰半乳糖胺构成,且其中总摩尔%为100%。
3.根据权利要求2所述的杂多糖,其特征在于:葡萄糖的摩尔%与半乳糖的摩尔%的比率在≥1.0和≤2.0之间。
4.根据权利要求2所述的杂多糖,其特征在于:其由40摩尔%的葡萄糖、20摩尔%的半乳糖、20摩尔%的鼠李糖和20摩尔%的N-乙酰半乳糖胺构成。
5.根据权利要求1-4中任一项所述的杂多糖,其特征在于:所述杂多糖包含重复单元,其中所述重复单元是由单糖葡萄糖和半乳糖和鼠李糖和N-乙酰半乳糖胺构成的五糖。
6.根据权利要求5所述的杂多糖,其中所述五糖由40摩尔%的葡萄糖、20摩尔%的半乳糖、20摩尔%的鼠李糖和20摩尔%的N-乙酰半乳糖胺构成。
7.能够产生权利要求1-6中任一项所限定的杂多糖的细菌。
8.根据权利要求7所述的细菌,其中所述细菌是Streptococcus salivariusthermophilus。
9.根据权利要求7所述的细菌,其选自由以下组成的组
·Streptococcus salivarius thermophilus NGB-22D,于2012年2月28日在乌特勒支的Centraalbureau voor Schimmelcultures(真菌生物多样性中心)保藏为CBS132067,
·Streptococcus salivarius thermophilus DS65008,于2013年6月18日在乌特勒支的Centraalbureau voor Schimmelcultures(真菌生物多样性中心)保藏为CBS135685。
10.一种生产发酵乳制品的方法,所述方法包括:添加权利要求1-6中任一项所限定的杂多糖或者添加权利要求7-9中任一项所限定的细菌。
11.根据权利要求10所述的方法,其中所述发酵乳制品选自由酸乳和奶酪组成的组。
12.一种生产权利要求1-6中任一项所限定的杂多糖的方法,所述方法包括:在有益于生产所述杂多糖的条件下培养权利要求7-9中所限定的乳酸细菌。
13.权利要求1-6中任一项所限定的杂多糖用于改善发酵乳制品的质地和/或粘度的用途。
14.权利要求7-9中所限定的乳酸细菌用于改善发酵乳制品的质地和/或粘度的用途。
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F.VANINGELGEM等: ""Biodiversity of Exopolysaccharides Produced by Streptococcus thermophilus Strains Is Reflected in Their Production and Their Molecular and Functional Characteristics"", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
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