CN114214377B - Phosphatidyl-agar oligosaccharide and preparation method thereof - Google Patents
Phosphatidyl-agar oligosaccharide and preparation method thereof Download PDFInfo
- Publication number
- CN114214377B CN114214377B CN202111600499.6A CN202111600499A CN114214377B CN 114214377 B CN114214377 B CN 114214377B CN 202111600499 A CN202111600499 A CN 202111600499A CN 114214377 B CN114214377 B CN 114214377B
- Authority
- CN
- China
- Prior art keywords
- phosphatidyl
- agar
- oligosaccharide
- phosphatidylcholine
- reaction
- 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.)
- Active
Links
- 229920001542 oligosaccharide Polymers 0.000 title claims abstract description 45
- 150000002482 oligosaccharides Chemical class 0.000 title claims abstract description 41
- 229920001817 Agar Polymers 0.000 title claims abstract description 40
- 239000008272 agar Substances 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 28
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims abstract description 24
- 230000002051 biphasic effect Effects 0.000 claims abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 239000006228 supernatant Substances 0.000 claims description 13
- 239000012074 organic phase Substances 0.000 claims description 11
- 229920000936 Agarose Polymers 0.000 claims description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 claims description 6
- 101710110830 Beta-agarase Proteins 0.000 claims description 4
- SKTCDJAMAYNROS-UHFFFAOYSA-N methoxycyclopentane Chemical compound COC1CCCC1 SKTCDJAMAYNROS-UHFFFAOYSA-N 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 claims description 4
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 claims description 3
- 238000007664 blowing Methods 0.000 claims description 3
- 239000000872 buffer Substances 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 235000021314 Palmitic acid Nutrition 0.000 claims description 2
- 230000000593 degrading effect Effects 0.000 claims description 2
- 235000020778 linoleic acid Nutrition 0.000 claims description 2
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 claims description 2
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 claims description 2
- JWMBOBQNPBCYER-UHFFFAOYSA-N 4-[(4,8-dihydroxy-2,6-dioxabicyclo[3.2.1]octan-3-yl)oxy]-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound OC1C(CO)OC(O)C(O)C1OC1C(O)C(C2O)OCC2O1 JWMBOBQNPBCYER-UHFFFAOYSA-N 0.000 claims 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims 2
- 229910052799 carbon Inorganic materials 0.000 claims 2
- 239000012071 phase Substances 0.000 claims 2
- 125000005313 fatty acid group Chemical group 0.000 claims 1
- 238000007789 sealing Methods 0.000 claims 1
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical group OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 abstract description 20
- 238000000034 method Methods 0.000 abstract description 16
- 238000005809 transesterification reaction Methods 0.000 abstract description 13
- 102000011420 Phospholipase D Human genes 0.000 abstract description 12
- 108090000553 Phospholipase D Proteins 0.000 abstract description 12
- 150000003904 phospholipids Chemical class 0.000 abstract description 9
- 230000015572 biosynthetic process Effects 0.000 abstract description 8
- -1 phosphatidyl glycoside Chemical class 0.000 abstract description 8
- 238000003786 synthesis reaction Methods 0.000 abstract description 7
- 229930182470 glycoside Natural products 0.000 abstract description 6
- 239000002502 liposome Substances 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 5
- 238000005538 encapsulation Methods 0.000 abstract description 4
- 238000011160 research Methods 0.000 abstract description 4
- 239000013543 active substance Substances 0.000 abstract description 2
- 238000006555 catalytic reaction Methods 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 235000010419 agar Nutrition 0.000 description 12
- 102000004190 Enzymes Human genes 0.000 description 9
- 108090000790 Enzymes Proteins 0.000 description 9
- 239000008346 aqueous phase Substances 0.000 description 8
- 239000000047 product Substances 0.000 description 7
- 239000000126 substance Substances 0.000 description 6
- 239000000758 substrate Substances 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 229930182830 galactose Natural products 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 230000003197 catalytic effect Effects 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 238000006116 polymerization reaction Methods 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000003892 spreading Methods 0.000 description 3
- 230000007480 spreading Effects 0.000 description 3
- 238000004809 thin layer chromatography Methods 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical group ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 125000003275 alpha amino acid group Chemical group 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 210000003050 axon Anatomy 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000002144 chemical decomposition reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 150000002016 disaccharides Chemical class 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000009144 enzymatic modification Effects 0.000 description 2
- 238000000105 evaporative light scattering detection Methods 0.000 description 2
- 229930182478 glucoside Natural products 0.000 description 2
- 150000002327 glycerophospholipids Chemical class 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 125000001095 phosphatidyl group Chemical group 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 150000004044 tetrasaccharides Chemical class 0.000 description 2
- HNAGHMKIPMKKBB-UHFFFAOYSA-N 1-benzylpyrrolidine-3-carboxamide Chemical compound C1C(C(=O)N)CCN1CC1=CC=CC=C1 HNAGHMKIPMKKBB-UHFFFAOYSA-N 0.000 description 1
- ZIIUUSVHCHPIQD-UHFFFAOYSA-N 2,4,6-trimethyl-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide Chemical compound CC1=CC(C)=CC(C)=C1S(=O)(=O)NC1=CC=CC(C(F)(F)F)=C1 ZIIUUSVHCHPIQD-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 1
- 208000017667 Chronic Disease Diseases 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
- LCGLNKUTAGEVQW-UHFFFAOYSA-N Dimethyl ether Chemical group COC LCGLNKUTAGEVQW-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 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
- 241000206672 Gelidium Species 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
- 102000015439 Phospholipases Human genes 0.000 description 1
- 108010064785 Phospholipases Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 108010045649 agarase Proteins 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 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 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000004641 brain development Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- OBNCKNCVKJNDBV-UHFFFAOYSA-N butanoic acid ethyl ester Natural products CCCC(=O)OCC OBNCKNCVKJNDBV-UHFFFAOYSA-N 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000002458 cell surface marker Substances 0.000 description 1
- 230000033077 cellular process Effects 0.000 description 1
- 230000021617 central nervous system development Effects 0.000 description 1
- 150000001793 charged compounds Chemical class 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 231100000481 chemical toxicant Toxicity 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 150000004665 fatty acids Chemical group 0.000 description 1
- 150000002190 fatty acyls Chemical group 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 1
- 238000005984 hydrogenation reaction Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 210000004498 neuroglial cell Anatomy 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose 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[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 150000003408 sphingolipids Chemical class 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000006277 sulfonation reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/12—Disaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
-
- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
- C12P7/6472—Glycerides containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
-
- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
- C12P7/6481—Phosphoglycerides
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Dispersion Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses phosphatidyl-agar oligosaccharides, which are prepared by the following method: the phosphatidylcholine and the agar oligosaccharide undergo transphosphatidylation in a biphasic reaction system under the catalysis of phospholipase D to synthesize phosphatidyl-agar oligosaccharide; the agaropectin oligosaccharide is selected from D-galactose and neoagaropectin; the phospholipase D is selected from PLDr34. The phosphatidyl-agar oligosaccharide provided by the invention is a novel phosphatidyl glycoside, and the phospholipid can be used for preparing liposome materials, and the transesterification product has an encapsulation function, so that the phosphatidyl-agar oligosaccharide provided by the invention has good application prospects in encapsulation and transportation, and can be used as an active substance for preparing liposomes, delivering medicines and the like. The invention explores the occurrence condition of transphosphatidylation of phosphatidylcholine and agaro-oligosaccharide, synthesizes novel phosphatidylglycoside, provides reasonable guidance for the synthesis of other phosphatidylglycoside, and has a certain research prospect.
Description
Technical Field
The invention relates to phosphatidyl-agar oligosaccharides and a preparation method thereof, belonging to the technical field of phosphatidyl glycosides.
Background
The ocean is a treasury and is a potential source of a plurality of natural bioactive substances. In all tissues, phospholipids are used as basic substances of life, and consist of hydrophilic head groups and hydrophobic tail groups, and can be divided into glycerophospholipids and sphingolipids due to different connection modes, wherein the content of Phosphatidylcholine (PC) in the glycerophospholipids is most abundant.
Phospholipids are not only capable of participating in the formation of cell membranes and maintaining their biological functionality, but also play an important role in cell micelles and organelles, for example, part of phospholipids are precursors of various anti-inflammatory compounds, which can regulate systemic inflammation and provide protection against chronic diseases. Phospholipids can be modified under a variety of conditions, such as hydrogenation, acetylation, sulfonation, and enzymatic modification, the first three methods of chemical modification being applied to the improvement of membrane physical properties, while enzymatic modification is a modification of hydrophilic head groups or hydrophobic fatty acyl chains, such as phospholipase D (PLD) -mediated transphosphatidylation. Several studies in animal models have shown that modified phospholipids have higher bioavailability and bioactivity, especially in terms of effects on plasma and liver lipid levels.
An important product of phospholipid modification is phosphatidylglycoside, and glycosylated phospholipids are mostly rich in lipid rafts or microdomains and play an important role in various cellular processes. In rodent brains, expression of phosphatidyl glucosides (ptdgc) is developmentally regulated. PtdGlc is most strongly expressed in radial glia at early brain development in rats and is considered a good cell surface marker for stem cells. Lysophosphatidyl glucosides (LPGlc) can act as guidance cues for the prolongation of axons during central nervous system development by activating the AG-like protein-coupled receptor (GPR) 55 of spinal sensory axons.
At present, the research on phosphatidyl glycoside is mainly focused on glucose, fructose, mannose and raffinose, and no intensive research on agar oligosaccharides exists.
The agar oligosaccharide is marine functional oligosaccharide with polymerization degree of 2-20 prepared by using agar as raw material. The preparation method of the agar oligosaccharides mainly comprises two methods of a chemical degradation method and a biological enzyme degradation method. The chemical method has many applications, but has some disadvantages, such as non-uniform product composition, complex operation, easy environmental pollution, etc. The biological enzyme degradation method is to hydrolyze the glycosidic bond on agarose chain by using specific agarase so as to obtain specific agaro-oligosaccharide, and has the advantages of high catalytic efficiency, good product specificity, mild reaction condition, no pollution and the like.
Disclosure of Invention
Aiming at the prior art, the invention provides a novel phosphatidyl glycoside-phosphatidyl-agar oligosaccharide and a preparation method thereof. The preparation method has the advantages of short reaction route, simple steps, high yield, no toxic chemicals involved in the synthesis process, and high stability of the transesterification product.
The invention is realized by the following technical scheme:
a phosphatidyl-agar oligosaccharide is prepared by the following method: the phosphatidylcholine and the agar oligosaccharide undergo transphosphatidylation in a biphasic reaction system under the catalysis of phospholipase D to synthesize phosphatidyl-agar oligosaccharide; the agaropectin oligosaccharide is selected from D-galactose and neoagaropectin; the phospholipase D is selected from PLDr34.
Further, the fatty acid chains on the phosphatidylcholine are palmitic acid of C16:0 and linoleic acid of C18:2, respectively.
Further, the phosphatidylcholine is dissolved in an organic solvent as an organic phase; the organic solvent is selected from methyl ether, diethyl ether, cyclopentyl methyl ether, ethyl acetate, butyl acetate or ethyl butyrate; the concentration of the phosphatidylcholine is 10-100 mg/mL.
Further, the agar oligosaccharides are dissolved in an aqueous solution to serve as an aqueous phase; the aqueous solution is selected from citric acid-sodium citrate buffer solution with pH value of 4.0-6.0.
The agar oligosaccharides can be prepared by a biological enzymolysis method or a chemical degradation method.
Further, the neoagarase is prepared by degrading agarose with beta agarase AgWH50B (Accession Number: KY 417136) and/or beta agarase AgWH50C (Accession Number: KC 913197) obtained by screening the subject group of the present inventors. The specific preparation method can be as follows: adding 2.0 g of AgWH50B crude enzyme into 1% agarose solution (g/ml) serving as a substrate, performing water bath reaction at 37 ℃ for 12 h and boiling water bath for 10 min, then adding 1.5 g of AgWH50C crude enzyme, performing water bath reaction at 37 ℃ for 12 h and boiling water bath for 10 min, centrifuging, taking supernatant, concentrating and drying to obtain the neoagalloch. The neoagalloch tetraose can be prepared by the following method: adding 2.0 g of AgWH50B crude enzyme into 1-2% agarose solution (g/ml) serving as a substrate, reacting in a water bath at 37 ℃ for 12 h, carrying out boiling water bath for 10 min, centrifuging to obtain a supernatant, concentrating and drying to obtain the neoagalloch tetrasaccharide.
Further, in the biphasic reaction system, the molar ratio of phosphatidylcholine to the agar oligosaccharides to each other is 1:45-55, preferably 1:50; the volume ratio of the organic phase to the aqueous phase is 1:0.8-1.2, preferably 1:1; the enzyme addition amount of the phospholipase D is 1.0-1.5U, preferably 1.4U.
Further, specific reaction conditions for the catalytic transphosphatidylation to occur are: the reaction system reacts in water bath at 37-42 ℃ for 8-12 h; centrifuging after the reaction is finished, taking supernatant, dissolving the product phosphatidyl-agar oligosaccharide in an upper organic phase, and blowing nitrogen to obtain the phosphatidyl-agar oligosaccharide.
The phosphatidyl-agar oligosaccharide provided by the invention is a novel phosphatidyl glycoside, and the phospholipid can be used for preparing liposome materials, and the transesterification product has an encapsulation function, so that the phosphatidyl-agar oligosaccharide provided by the invention can be predicted to have good application prospects in encapsulation and transportation, and can be used as an active substance (such as a drug carrier, a liposome membrane material and the like) for preparing liposome, drug delivery and the like.
The invention explores the occurrence condition of transphosphatidylation of phosphatidylcholine and agar oligosaccharides, connects the screened agar oligosaccharides with higher substrate preference to phosphatidylcholine through transphosphatidylation, synthesizes a novel phosphatidyl glycoside-phosphatidyl-agar oligosaccharides, provides reasonable guidance for the synthesis of other phosphatidyl glycosides, and has a certain research prospect. The preparation method of the invention adopts double-enzyme synthesis, thus greatly reducing the production cost.
The various terms and phrases used herein have the ordinary meaning known to those skilled in the art.
Drawings
Fig. 1: TLC results of transesterification of phosphatidylcholine with agaro-oligosaccharides with different degrees of polymerization are shown, wherein PC represents phosphatidylcholine, and 1, 2, 3 and 4 represent transesterification results of PC with D-galactose, neoagaronose and 1-15AOS respectively.
Fig. 2: chemical structure of phosphatidylcholine.
Fig. 3: chemical structure of phosphatidyl-D galactose.
Fig. 4: chemical structure of phosphatidyl-neoagalloch.
Fig. 5: mass spectrometry results of phosphatidyl-D galactose are schematically shown.
Fig. 6: the mass spectrum analysis result of phosphatidyl-neoagalloch disaccharide is shown schematically.
Fig. 7: results of HPLC-ELSD analysis of phosphatidyl-agar oligosaccharides.
Detailed Description
The invention is further illustrated below with reference to examples. However, the scope of the present invention is not limited to the following examples. Those skilled in the art will appreciate that various changes and modifications can be made to the invention without departing from the spirit and scope thereof.
The instruments, reagents, materials, etc. used in the examples described below are conventional instruments, reagents, materials, etc. known in the art, and are commercially available. The experimental methods, detection methods, and the like in the examples described below are conventional experimental methods, detection methods, and the like that are known in the prior art unless otherwise specified.
The phosphatidylcholine used in the present invention is Soy PC (95%) (SPC, 441601G-50G-I-175,Avanti Polar Lipids).
The 1-15AOS used in the invention is mixed sugar of agarose monosaccharide and pentadecyl saccharide, and 1-15 agar oligosaccharide (1500 Da, qingdao Bozhi Hui Biotechnology Co., ltd.).
The phospholipase D used in the invention is PLDr34 (Accession Number: MN 604233) (this enzyme is described in another patent application of the applicant of the invention, CN 110564708A, i.e., phospholipase shown as SEQ ID NO.1 in the specification of CN 110564708A), and its amino acid sequence is shown below.
Amino acid sequence of PLDr 34:
MIISFRLSRPARAALICALALTVLPASPATAADAATPHLDAVERTLREVSPGLEGEVWERTAGNRLDAGADDPAGWLLQTPGCWGDAGCRDRVGTRRLLAKMTENISRATRTVDISTLAPFPNGAFQDAIVAGLKSSAARGNKLTVRVLVGAAPIYHMNVLPSKYRDELVAKLGADARNVDLNVASMTTSKTSFSWNHSKLLVVDGQSVITGGINDWKDDYLETAHPVADVDLALRGPAAASAGRYLDELWSWTCQNRNNIAGVWFASSNGTACMPAMAKDTAPAAPPAAPGDVPAIAVGGLGVGIKRSDPSSAFRPTLPSAADTKCVVGLHDNTNADRDYDTVNPEESALRTLISSAKGHIEISQQDVNATCPPLPRYDIRVYDALAARMAAGVKVRIVVSDPANRGAVGSGGYSQIKSLSEISDTLRDRLALLTGDQGAAKATMCSNLQLATFRSSKSPTWADGHPYAQHHKVVSVDDSAFYIGSKNLYPAWLQDFGYIVESPGAAQQLDAQLLSPQWTHSKETATVDYERGLCHI。
experiment 1: substrate preference selection for transphosphorylation reactions
The method comprises the following steps:
(one) transphospholipid acylation reaction
(1) 10 mg phosphatidylcholine is dissolved in 1 mL cyclopentyl methyl ether to obtain an organic phase.
(2) 0.12 g of D-galactose was dissolved in 1 mL citric acid-sodium citrate buffer (pH 6.0, 0.1M) to obtain an aqueous phase.
(3) The organic and aqueous phases were mixed in a volume ratio of 1:1, PLD 1.4. 1.4U was added to make up the biphasic reaction conditions and sealed in a brown vial.
(4) The brown vials were placed in a constant temperature water bath and reacted in a water bath at 40℃and 200 r for 10 h.
(5) After the reaction, the mixture was centrifuged (8000 r,5 min) to obtain a supernatant A.
D-galactose in the step (2) is respectively replaced by 0.21 g neoagalloch, 0.49 g neoagalloch, 1.5 g 1-15 agalloch oligosaccharides (other conditions are the same), and supernatant B, supernatant C and supernatant D are obtained.
(II) performing thin layer chromatography on the supernatant A, the supernatant B, the supernatant C and the supernatant D prepared above:
(1) Samples were sampled with a capillary volume of 0.5mm, and dried with a blower after each sample application.
(2) Placing the silica gel plate with the sample in a spreading cylinder containing a spreading agent, wherein the formula of the spreading agent is chloroform: methanol: glacial acetic acid: water=50:25:6:2 (v: v: v).
(3) After about 20 min, the silica gel plate is taken out, and after the silica gel plate is dried by a blower, the silica gel plate is put into an iodine jar, and the experimental result is observed later.
As can be seen from the thin-layer chromatography analysis result graph (shown in figure 1), in the transphosphorylation reaction taking the agaro-oligosaccharide with different polymerization degrees as the raw material, the transesterification reaction involving the neoagaro-disaccharide is more thoroughly carried out, and the substrate PC is almost completely degraded. The transesterification reaction with D-galactose is also more thorough, the transesterification reaction with neoagaragar tetrasaccharide is weaker, and the transesterification reaction with mixed agaragar oligosaccharide does not occur. The transphosphatidylated agaropectin oligosaccharides were subsequently identified as D-galactose and neoagaropectin.
Example 1: PLD catalytic enzyme synthesis of phosphatidyl-agar oligosaccharides
The method comprises the following steps:
(1) 10 mg phosphatidylcholine is dissolved in 1 mL cyclopentyl methyl ether to obtain an organic phase.
(2) 0.12 g of D-galactose was dissolved in 1 mL citric acid-sodium citrate buffer (pH 6.0, 0.1M) to obtain an aqueous phase.
(3) The organic and aqueous phases were mixed in a volume ratio of 1:1, PLD 1.4. 1.4U was added to make up the biphasic reaction conditions and sealed in a brown vial.
(4) The brown vials were placed in a constant temperature water bath and reacted in a water bath at 40℃and 200 r for 10 h.
(5) After the reaction, centrifuging (8000 r,5 min) to obtain supernatant, and obtaining phosphatidyl-agar oligosaccharide dissolved in organic solvent.
(6) After nitrogen blowing, the phosphatidyl-agar oligosaccharides with high concentration are obtained, and the structural formula is shown in figure 3.
And (3) replacing D-galactose in the step (2) with 0.21 g neoagalloch (other conditions are the same), and preparing the phosphatidyl-neoagalloch, wherein the structural formula of the phosphatidyl-neoagalloch is shown in figure 4.
The relative molecular masses of the two phosphatidyl-agaragar oligosaccharides were measured by MS method, and the results are shown in FIG. 5 and FIG. 6, and the analysis results are consistent with the calculation results of the corresponding molecular ion peaks (M phosphatidyl-D galactose. Apprxeq.851, M phosphatidyl-neoagarobiose. Apprxeq.995), and the results of mass spectrometry analysis results and the results of the liquid-phase product peaks both indicate that the target phosphatidyl glycoside is formed after transesterification reaction, so that the phosphatidyl-D galactose and the phosphatidyl-neoagarobiose are determined to be produced.
Experiment 2: key parameters of transphosphatidyl preparation process
Parameters critical to the transphosphorylation reaction include: in the transesterification reaction, molar ratio of PC to sugar (1:20, 1:30, 1:40, 1:50, 1:60), PLD enzyme addition (0.2, 0.6, 1, 1.4, 1.8, U), transesterification reaction time (4, 6, 8, 10, 12 h) and volume ratio of organic phase to aqueous phase (1:3, 1:2, 1:1, 2:1, 3:1) in the biphasic reaction system. The 4 factors in the reaction system are regulated and controlled, and the synthesis conversion rate is detected by adopting an HPLC-ELSD method, so that the optimal synthesis condition of the phosphatidyl-agar oligosaccharides is finally determined. When the molar ratio of PC to sugar was 1:50, the PLD-added enzyme amount was 1.4: 1.4U, the transesterification reaction time was 10: 10 h, and the volume ratio of the organic phase to the aqueous phase was 1:1, the conversion of phosphatidyl-D-galactose could reach 85% and the conversion of phosphatidyl-neoagalloch disaccharide could reach 96% under the optimal reaction conditions (FIG. 7). Experimental results show that when the transphosphatidylation reaction is carried out by taking neoagalloch as a raw material, the conversion rate of phosphatidyl-neoagalloch is obviously higher than that of D-galactose.
The foregoing examples are provided to fully disclose and describe how to make and use the claimed embodiments by those skilled in the art, and are not intended to limit the scope of the disclosure herein. Modifications that are obvious to a person skilled in the art will be within the scope of the appended claims.
Claims (1)
1. The preparation method of the phosphatidyl-agar oligosaccharides is characterized by comprising the following steps:
(1) Dissolving 10 mg phosphatidylcholine in 1 mL cyclopentyl methyl ether to obtain an organic phase;
(2) Dissolving 0.21 g neoagalloch in 1 mL pH 6.0, 0.1M citric acid-sodium citrate buffer to obtain water phase;
(3) Mixing the organic phase and the water phase according to the volume ratio of 1:1, adding PLDr34 of 1.4U to form a biphasic reaction condition, and sealing in a brown vial;
(4) Placing the brown vial into a constant temperature water bath kettle, and carrying out water bath reaction under the conditions of 40 ℃ and 200 r for 10 h;
(5) Centrifuging at 8000 r for 5 min after the reaction is finished to obtain supernatant, and obtaining phosphatidyl-agar oligosaccharides dissolved in an organic solvent;
(6) After nitrogen blowing, the phosphatidyl-agar oligosaccharides are obtained;
the fatty acid chains on the phosphatidylcholine are palmitic acid with the carbon number of 16:0 and linoleic acid with the carbon number of 18:2 respectively;
the neoagarobiose is prepared by degrading agarose by beta agarase AgWH50B and/or beta agarase AgWH 50C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111600499.6A CN114214377B (en) | 2021-12-24 | 2021-12-24 | Phosphatidyl-agar oligosaccharide and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111600499.6A CN114214377B (en) | 2021-12-24 | 2021-12-24 | Phosphatidyl-agar oligosaccharide and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114214377A CN114214377A (en) | 2022-03-22 |
| CN114214377B true CN114214377B (en) | 2024-03-08 |
Family
ID=80705935
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111600499.6A Active CN114214377B (en) | 2021-12-24 | 2021-12-24 | Phosphatidyl-agar oligosaccharide and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114214377B (en) |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4624919A (en) * | 1983-04-11 | 1986-11-25 | Meito Sangyo Kabushiki Kaisha | Enzymatic production of phospholipid-saccharide derivatives |
| JPH0656856A (en) * | 1993-04-16 | 1994-03-01 | Meito Sangyo Kk | Phospholipid derivative |
| JPH07188274A (en) * | 1993-12-27 | 1995-07-25 | Honen Corp | Novel galactose derivative |
| JPH10226616A (en) * | 1997-02-18 | 1998-08-25 | Noevir Co Ltd | Skin preparation for external use |
| WO2000018945A1 (en) * | 1998-10-01 | 2000-04-06 | Albany Molecular Research, Inc. | Transphosphatidylation catalized by phospholipase d in anhydrous organic solvents in the presence of ion-exchange resin |
| WO2015041498A1 (en) * | 2013-09-23 | 2015-03-26 | 다인바이오 주식회사 | Immune enhancement or anticancer composition containing neoagarooligosaccharide |
| CN105255967A (en) * | 2015-11-12 | 2016-01-20 | 福州大学 | Enzymolysis preparation method of new agaro oligosaccharides |
| KR20180041377A (en) * | 2016-10-14 | 2018-04-24 | 명지대학교 산학협력단 | A Novel alpha-neoagarobiose hydrolase from Gayadomonas joobiniege G7 and use thereof |
| CN110564708A (en) * | 2019-10-19 | 2019-12-13 | 中国海洋大学 | Recombinant phospholipase D and application thereof in synthesis of phosphatidylserine or other phospholipids |
| WO2020133314A1 (en) * | 2018-12-29 | 2020-07-02 | 邦泰生物工程(深圳)有限公司 | Phospholipase d mutant, use thereof and method for preparing phosphatidylserine by means of same |
| CN111514152A (en) * | 2020-05-18 | 2020-08-11 | 中国海洋大学 | Application of n-3PUFA phosphatidyl glycoside in preparation for improving blood brain barrier damage |
| CN111621535A (en) * | 2020-05-18 | 2020-09-04 | 中国海洋大学 | Preparation method and application of phosphatidyl glycoside |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090238811A1 (en) * | 2002-09-09 | 2009-09-24 | Mcdaniel C Steven | Enzymatic Antimicrobial and Antifouling Coatings and Polymeric Materials |
| WO2005123766A2 (en) * | 2004-06-09 | 2005-12-29 | Alexander Sunguroff | Methods of making nanotechnological and macromolecular biomimetic structures |
| KR101521711B1 (en) * | 2013-10-14 | 2015-05-19 | 고려대학교 산학협력단 | Novel β-agarooligosaccharide hydrolase and enzymatic production method of 3,6-anhydro-L-galactose and galactose from agarose by using the same |
-
2021
- 2021-12-24 CN CN202111600499.6A patent/CN114214377B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4624919A (en) * | 1983-04-11 | 1986-11-25 | Meito Sangyo Kabushiki Kaisha | Enzymatic production of phospholipid-saccharide derivatives |
| JPH0656856A (en) * | 1993-04-16 | 1994-03-01 | Meito Sangyo Kk | Phospholipid derivative |
| JPH07188274A (en) * | 1993-12-27 | 1995-07-25 | Honen Corp | Novel galactose derivative |
| JPH10226616A (en) * | 1997-02-18 | 1998-08-25 | Noevir Co Ltd | Skin preparation for external use |
| WO2000018945A1 (en) * | 1998-10-01 | 2000-04-06 | Albany Molecular Research, Inc. | Transphosphatidylation catalized by phospholipase d in anhydrous organic solvents in the presence of ion-exchange resin |
| WO2015041498A1 (en) * | 2013-09-23 | 2015-03-26 | 다인바이오 주식회사 | Immune enhancement or anticancer composition containing neoagarooligosaccharide |
| CN105255967A (en) * | 2015-11-12 | 2016-01-20 | 福州大学 | Enzymolysis preparation method of new agaro oligosaccharides |
| KR20180041377A (en) * | 2016-10-14 | 2018-04-24 | 명지대학교 산학협력단 | A Novel alpha-neoagarobiose hydrolase from Gayadomonas joobiniege G7 and use thereof |
| WO2020133314A1 (en) * | 2018-12-29 | 2020-07-02 | 邦泰生物工程(深圳)有限公司 | Phospholipase d mutant, use thereof and method for preparing phosphatidylserine by means of same |
| CN110564708A (en) * | 2019-10-19 | 2019-12-13 | 中国海洋大学 | Recombinant phospholipase D and application thereof in synthesis of phosphatidylserine or other phospholipids |
| CN111514152A (en) * | 2020-05-18 | 2020-08-11 | 中国海洋大学 | Application of n-3PUFA phosphatidyl glycoside in preparation for improving blood brain barrier damage |
| CN111621535A (en) * | 2020-05-18 | 2020-09-04 | 中国海洋大学 | Preparation method and application of phosphatidyl glycoside |
Non-Patent Citations (1)
| Title |
|---|
| 磷脂酶D的细胞信号转导作用;钟秀丽;《植物生理与分子生物学学报》(第5期);第451-460页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114214377A (en) | 2022-03-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Galan et al. | Recent developments of ionic liquids in oligosaccharide synthesis: the sweet side of ionic liquids | |
| DE60031957T2 (en) | COMBINATIVE, COMPLEX CARBOHYDRATE LIBRARIES AND METHOD FOR THE PREPARATION AND USE THEREOF | |
| KR0147845B1 (en) | Glycosphingolipids with a group capable of coupling in the sphingoid portion, the preparation and use thereof | |
| Thiem et al. | Chemoenzymatic syntheses of sialyloligosaccharides with immobilized sialidase | |
| Fauré et al. | Synthesis of a library of xylogluco-oligosaccharides for active-site mapping of xyloglucan endo-transglycosylase | |
| CN107034205B (en) | Method for preparing high-esterification-activity lipase by using surfactant | |
| Galan et al. | Ionic-liquid-based MS probes for the chemo-enzymatic synthesis of oligosaccharides | |
| Yano et al. | Total synthesis of Myc-IV (C16: 0, S) via automated electrochemical assembly | |
| JPH07500494A (en) | Enzymatic synthesis of carbohydrates | |
| KR20000067921A (en) | Process for the preparation of sphingolipids and sphingolipid derivatives | |
| US20030119051A1 (en) | Saccharide library | |
| CN114214377B (en) | Phosphatidyl-agar oligosaccharide and preparation method thereof | |
| Filice et al. | Monosaccharide derivatives as central scaffolds in the synthesis of glycosylated drugs | |
| Sandoval et al. | Development of regioselective deacylation of peracetylated β-d-monosaccharides using lipase from Pseudomonas stutzeri under sustainable conditions | |
| Filice et al. | Preparation of linear oligosaccharides by a simple monoprotective chemo-enzymatic approach | |
| Gao et al. | Regioselective synthesis of dimeric (Gemini) and trimeric sugar‐based surfactants | |
| EP0906324B1 (en) | Carbohydrate derivatives and their solid-phase synthesis | |
| Bavaro et al. | Flow-based biocatalysis: Application to peracetylated arabinofuranosyl-1, 5-arabinofuranose synthesis | |
| Zheng et al. | Chemoenzymatic Synthesis of Glycoconjugates Mediated by Regioselective Enzymatic Hydrolysis of Acetylated 2‐Amino Pyranose Derivatives | |
| Quesada Allue et al. | Lipid‐bound oligosaccharides in insects | |
| JP2017516838A (en) | Hexose derivatives, their formulation and use | |
| CN114410717A (en) | Phosphatidyl glucoside and preparation method and application thereof | |
| 安藤弘宗 et al. | A synthetic challenge to the diversity of gangliosides for unveiling their biological significance | |
| Yamazaki et al. | Preparation and characterization of neoglycoprotein-liposome conjugates: A promising approach to developing drug delivery materials applying sugar chain ligands | |
| FR3110176A1 (en) | Strain and process for the production of oligosaccharides |
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 |