EP3989731A1 - Process for the production of rubisco from a plant biomass - Google Patents
Process for the production of rubisco from a plant biomassInfo
- Publication number
- EP3989731A1 EP3989731A1 EP20747040.2A EP20747040A EP3989731A1 EP 3989731 A1 EP3989731 A1 EP 3989731A1 EP 20747040 A EP20747040 A EP 20747040A EP 3989731 A1 EP3989731 A1 EP 3989731A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- rubisco
- carried out
- juice
- solution
- dehydration
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108010003581 Ribulose-bisphosphate carboxylase Proteins 0.000 title claims abstract description 139
- 238000000034 method Methods 0.000 title claims abstract description 68
- 239000002028 Biomass Substances 0.000 title claims abstract description 25
- 238000004519 manufacturing process Methods 0.000 title description 18
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000003456 ion exchange resin Substances 0.000 claims abstract description 10
- 229920003303 ion-exchange polymer Polymers 0.000 claims abstract description 10
- 239000011347 resin Substances 0.000 claims description 72
- 229920005989 resin Polymers 0.000 claims description 72
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 58
- 235000015192 vegetable juice Nutrition 0.000 claims description 45
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 41
- 230000018044 dehydration Effects 0.000 claims description 34
- 238000006297 dehydration reaction Methods 0.000 claims description 34
- 238000010828 elution Methods 0.000 claims description 21
- 238000000108 ultra-filtration Methods 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 238000000889 atomisation Methods 0.000 claims description 17
- 238000005119 centrifugation Methods 0.000 claims description 16
- 230000003750 conditioning effect Effects 0.000 claims description 16
- 230000014759 maintenance of location Effects 0.000 claims description 16
- 238000003825 pressing Methods 0.000 claims description 16
- 238000004587 chromatography analysis Methods 0.000 claims description 15
- 125000000129 anionic group Chemical group 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 14
- 238000001694 spray drying Methods 0.000 claims description 10
- 229920000936 Agarose Polymers 0.000 claims description 9
- 239000011159 matrix material Substances 0.000 claims description 9
- 238000011067 equilibration Methods 0.000 claims description 8
- 229920000642 polymer Polymers 0.000 claims description 7
- 235000013311 vegetables Nutrition 0.000 claims description 7
- 229920006037 cross link polymer Polymers 0.000 claims description 6
- 238000011026 diafiltration Methods 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 125000001453 quaternary ammonium group Chemical group 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 150000004676 glycans Chemical class 0.000 claims description 5
- 229920001282 polysaccharide Polymers 0.000 claims description 5
- 239000005017 polysaccharide Substances 0.000 claims description 5
- 239000002131 composite material Substances 0.000 claims description 4
- 238000001542 size-exclusion chromatography Methods 0.000 claims description 4
- 229920002307 Dextran Polymers 0.000 claims description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- 230000001174 ascending effect Effects 0.000 claims description 3
- 239000000872 buffer Substances 0.000 claims description 3
- 239000008188 pellet Substances 0.000 claims description 3
- 229910052814 silicon oxide Inorganic materials 0.000 claims description 3
- 150000003512 tertiary amines Chemical class 0.000 claims description 3
- 239000008121 dextrose Substances 0.000 claims description 2
- 125000000524 functional group Chemical group 0.000 claims description 2
- 229930182830 galactose Natural products 0.000 claims description 2
- QOHMWDJIBGVPIF-UHFFFAOYSA-N n',n'-diethylpropane-1,3-diamine Chemical group CCN(CC)CCCN QOHMWDJIBGVPIF-UHFFFAOYSA-N 0.000 claims 1
- 238000013375 chromatographic separation Methods 0.000 abstract description 3
- YAHZABJORDUQGO-NQXXGFSBSA-N D-ribulose 1,5-bisphosphate Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)C(=O)COP(O)(O)=O YAHZABJORDUQGO-NQXXGFSBSA-N 0.000 abstract description 2
- 102000004020 Oxygenases Human genes 0.000 abstract description 2
- 108090000417 Oxygenases Proteins 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 39
- 241000219823 Medicago Species 0.000 description 37
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 description 35
- 241000196324 Embryophyta Species 0.000 description 28
- 235000018102 proteins Nutrition 0.000 description 21
- 108090000623 proteins and genes Proteins 0.000 description 21
- 102000004169 proteins and genes Human genes 0.000 description 21
- 229920002684 Sepharose Polymers 0.000 description 13
- 239000012528 membrane Substances 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 12
- 238000001914 filtration Methods 0.000 description 11
- 239000000463 material Substances 0.000 description 11
- 239000007788 liquid Substances 0.000 description 10
- 239000012466 permeate Substances 0.000 description 10
- 229930002875 chlorophyll Natural products 0.000 description 7
- 235000019804 chlorophyll Nutrition 0.000 description 7
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 7
- 239000000284 extract Substances 0.000 description 7
- 238000000926 separation method Methods 0.000 description 7
- 239000013622 capto Q Substances 0.000 description 6
- 239000012465 retentate Substances 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 150000003839 salts Chemical class 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 238000004132 cross linking Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000008055 phosphate buffer solution Substances 0.000 description 4
- 239000011148 porous material Substances 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000001143 conditioned effect Effects 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 235000015032 reconstituted 100% juice Nutrition 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- OSJPPGNTCRNQQC-UWTATZPHSA-N 3-phospho-D-glyceric acid Chemical compound OC(=O)[C@H](O)COP(O)(O)=O OSJPPGNTCRNQQC-UWTATZPHSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 229920012266 Poly(ether sulfone) PES Polymers 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O ammonium group Chemical group [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 210000003763 chloroplast Anatomy 0.000 description 2
- 238000005345 coagulation Methods 0.000 description 2
- 230000015271 coagulation Effects 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000005342 ion exchange Methods 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 230000002211 methanization Effects 0.000 description 2
- 230000002572 peristaltic effect Effects 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 230000000243 photosynthetic effect Effects 0.000 description 2
- 150000008442 polyphenolic compounds Chemical class 0.000 description 2
- 235000013824 polyphenols Nutrition 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000007787 solid Chemical group 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- ASCFNMCAHFUBCO-UHFFFAOYSA-N 2-phosphoglycolic acid Chemical compound OC(=O)COP(O)(O)=O ASCFNMCAHFUBCO-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 240000007124 Brassica oleracea Species 0.000 description 1
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 description 1
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 description 1
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 241000206672 Gelidium Species 0.000 description 1
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 241000209082 Lolium Species 0.000 description 1
- 240000004658 Medicago sativa Species 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 240000006394 Sorghum bicolor Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 244000300264 Spinacia oleracea Species 0.000 description 1
- 235000009337 Spinacia oleracea Nutrition 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 241000219793 Trifolium Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 235000010419 agar Nutrition 0.000 description 1
- 230000002009 allergenic effect Effects 0.000 description 1
- 150000001412 amines Chemical group 0.000 description 1
- 244000062766 autotrophic organism Species 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 238000010908 decantation Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- -1 diethylaminopropyl Chemical group 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000001177 diphosphate Substances 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000002663 nebulization Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 235000002949 phytic acid Nutrition 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000010287 polarization Effects 0.000 description 1
- 229920000058 polyacrylate Polymers 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 235000012045 salad Nutrition 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 238000003998 size exclusion chromatography high performance liquid chromatography Methods 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
-
- 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
- C12N9/88—Lyases (4.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y401/00—Carbon-carbon lyases (4.1)
- C12Y401/01—Carboxy-lyases (4.1.1)
- C12Y401/01039—Ribulose-bisphosphate carboxylase (4.1.1.39)
Definitions
- the present invention relates to a process for producing ribulose-l, 5-biphosphate carboxylase-oxygenase, also called Rubisco, from a plant biomass.
- Rubisco is a plant enzyme involved in the Calvin cycle and more particularly in the binding of carbon dioxide.
- Rubisco located in chloroplasts, is one of the most abundant proteins in plant biomass and even the most abundant on earth. This enzyme is involved in the process of photosynthesis and carries two enzymatic activities: i) a carboxylase activity during which Rubisco fixes a CO2 on D-ribulose-l, 5-diphosphate to form two molecules of 3-phospho-D -glycerate, and ii) oxygenase activity during which Rubisco binds an O 2 molecule to D-ribulose-1,5-diphosphate to form 3-phospho-D-glycerate and 2-phosphoglycolate.
- Rubisco is also renowned for its nutritional qualities and is low allergenic. Indeed, Rubisco contains an amino-gram equivalent to that of milk or egg proteins, which places it advantageously in front of other proteins of plant origin. In addition, various studies have highlighted the functional properties of Rubisco such as its emulsifying, foaming and gelling power, which, associated with good solubility, has aroused a strong interest in its use in new applications with high added value. .
- patent application FR 2 819 685 describes a process for treating the green juice resulting from the pressing of a leaf material rich in proteins such as alfalfa, in order to recover therefrom products having a strong interest in food, in particular for humans.
- This process comprises a) a separation step to obtain, on the one hand, a brown juice rich in proteins
- cytoplasmic and, on the other hand, a fraction rich in particular in proteins, pigments, vitamins, insolubles and trace elements, then b) drying this fraction resulting from step a) to obtain a product rich in particular in proteins, vitamins and trace elements. elements.
- this process it is then possible to subject the brown juice to a separation on resin in order to recover a decolored juice rich in proteins and a polyphenolic extract.
- This process comprises a very large number of steps and is complex to implement. In addition, it does not allow access to pure Rubisco. Indeed, the juice obtained contains a mixture of different proteins, and polyphenols.
- a liquid juice is obtained containing the Rubisco which is then subjected to a heat treatment at a temperature of 50 to 80 ° C to cause the coagulation of the protein which is then recovered, by for example by centrifugation or decantation of the juice, and filtration.
- This process does not necessarily lead to pure Rubisco as the squeezed juice may contain other proteins.
- the coagulation stage leads to the denaturation of the protein (s) present in the liquid juice, which can alter their functional properties.
- the inventors have therefore set themselves the goal of providing a process for the production and purification of Rubisco which is both simple and economical to implement, transposable to an industrial scale, to lead to the production, with high yields, of a purified Rubisco whose functional properties are not altered.
- the subject of the present invention is therefore a process for the production of purified Rubisco from plant biomass, said process comprising, in this order, at least the following stages:
- step 2) a step of reconstituting a vegetable juice from the powder obtained in step 1), to lead to a reconstituted vegetable juice
- step 3 a step of centrifugation of the reconstituted vegetable juice obtained above in step 2), to lead to a centrifuged juice and to a centrifugation pellet,
- step 4) of chromatographic retention is carried out in a fixed bed or in an expanded bed or in a fluidized bed, in a chromatography column containing an ion exchange resin chosen from anionic hydrophilic resins containing functional groups chosen from tertiary amines and quaternary ammoniums, said resins comprising i) either a matrix of at least one crosslinked polymer, ii) or a composite matrix of silicon oxide and of at least one polyacrylic polymer.
- an ion exchange resin chosen from anionic hydrophilic resins containing functional groups chosen from tertiary amines and quaternary ammoniums
- the term “purified Rubisco” is understood to mean a Rubisco free from other macromolecules (analysis by SEC HPLC at 280 nm) but which may nevertheless be in the form of a mixture with compounds such as salts, water or other small molecules. In general, these small molecules are less than 1 kDa in size and cannot be called proteins. These small molecules ensure the stability and / or solubility of Rubisco. These other compounds represent from 1 to 5% by maximum mass relative to the total mass of Rubisco + other compounds. Thus, according to the invention, a purified Rubisco is a 95-99% pure Rubisco.
- the anionic hydrophilic resins which can be used according to the process in accordance with the present invention may have variable pore diameters but preferably have a diameter sufficient to allow good diffusion of the Rubisco throughout the resin during chromatographic retention step 4). .
- the pore diameter of the anionic hydrophilic resins is preferably between 20 and 100 nm approximately, and even more
- the resins comprising a crosslinked polymer matrix are chosen from resins in which said crosslinked polymer is a polysaccharide, in particular a galactose polymer, very particularly agarose (or agar-agar ) or branched polymer of dextrose (glucose), most particularly dextran.
- a galactose polymer very particularly agarose (or agar-agar ) or branched polymer of dextrose (glucose), most particularly dextran.
- this type of resin makes it possible to lead to yields of purified Rubisco of the order of 20 to 40 g / L of resin used.
- the resins mentioned in points a) and c) are very particularly preferred, these leading to yields of purified Rubisco of the order of 7 to 9 g, respectively of 4 to 6 g, of purified Rubisco. / L of resin used.
- All the leafy plants producing Rubisco can be used as plant biomass according to the process of the invention.
- the plant biomass used to obtain the green juice from step 1) can in particular come from a plant cultivated specifically to produce Rubisco, such as a fodder plant such as alfalfa, clover, ryegrass, sorghum forage, etc ...; a vegetable plant such as cabbage, spinach, salad; the tobacco ; a succulent plant; and more generally all plants having a high leaf / plant ratio.
- alfalfa is very particularly preferred.
- the process according to the invention opens, in this case, other outlets for the agricultural sectors.
- Plant biomass can also be a co-product of an industry or a green waste. In this case, the process according to the invention allows material recovery from sectors already in place or makes it possible to develop new sectors (for example the recovery of grass clippings which can be recovered and constitute plant biomass).
- the plant biomass containing Rubisco is preferably fresh biomass.
- the biomass used in step 1) has just been collected or picked.
- coarse pre-grinding may be useful before pressing.
- Pressing is usually done in a screw press that operates continuously. At the end of the pressing, a vegetable juice is obtained which is a complex solution containing Rubisco and other compounds such as many polysaccharides and a solid residue (pressing cake) which can be upgraded, for example, by methanization or be used for animal feed.
- the pH of the vegetable juice resulting from the pressing can be adjusted between 6.5 and 7.5 if necessary in order to ensure better stability of the Rubisco during the following steps of the process.
- Dehydration step 1) can be carried out at a temperature ranging from 58 to 200 ° C approximately, preferably ranging from 60 to 150 ° C approximately, and particularly preferably ranging from 65 to 130 ° C approximately.
- step 1) of dehydration is carried out at a temperature less than or equal to 140 ° C, and even more preferably at a temperature less than or equal to 120 ° C.
- the total duration of dehydration step 1) is less than approximately 20 seconds.
- Step 1) of dehydration of the vegetable juice can in particular be carried out by a process of drying by atomization, lyophilization or evapoconcentration.
- step 1) of dehydration is carried out by spray drying.
- This process consists in carrying out a nebulization or dispersion (also called atomization) of the vegetable juice in liquid droplets which are brought into contact with a current of hot air which provides the energy necessary for the evaporation and transport of the solvent.
- a nebulization or dispersion also called atomization
- a large number of non-independent parameters influence the properties of the dry powder, in particular, variables related to the drying process and variables related to the formulation or to the
- composition of the liquid charge is a composition of the liquid charge.
- step 1) of dehydration is carried out by spray drying at an inlet temperature into the atomizer (first temperature) ranging from 110 to less than approximately 200 ° C, from particularly preferably at an inlet temperature ranging from 120 to 150 ° C approximately, and more particularly preferably at an inlet temperature ranging from 120 to 130 ° C approximately; followed by spray drying at an atomizer outlet temperature (second temperature) ranging from approximately 58 to 95 ° C, particularly preferably at a second temperature ranging from approximately 60 to 75 ° C, and more particularly preferred at a second temperature ranging from 65 to 75 ° C approximately.
- first temperature inlet temperature into the atomizer
- the duration of exposure to the inlet temperature is less than or equal to 2 seconds and the total duration of the dehydration step.
- dehydration is less than about 15 seconds. Even more preferably, the exposure time to the input temperature varies from l / 10th of a second to 1 second and the total duration of the dehydration step is less than approximately 12 seconds.
- the process can for example be carried out in a laboratory atomizer-dryer such as the device sold under the trade name Mini Büchi Spray-Dryer B-290 by the company Büchi or in a pilot atomizer dryer sold under the trade name Niro Minor SD 2 by the company GEA.
- the dryer can have an inlet temperature corresponding to the first temperature as defined in the invention, and an outlet temperature corresponding to the second temperature as defined in 'invention.
- step 1) of dehydration carried out under the conditions mentioned above guarantees the stability of the Rubisco and facilitates storage by reducing the volume. It is thus possible to store the dried juices to overcome the periods of harvesting plant biomass and better distribute the production of Rubisco throughout the year.
- this dehydration step 1) has a positive effect on the progress of the subsequent chromatographic retention step 4). It is also possible to mix powders from different juices (different batches, different plant origin, etc.).
- Step 2) of reconstituting a vegetable juice can be carried out by adding water or a solution making it possible to adjust the pH or the salinity of the juice if necessary.
- the volume of water or solution added to the powder (dehydrated vegetable juice) during step 2) is preferably equal to the volume of the vegetable juice resulting from the pressing of the vegetable biomass before its dehydration.
- Step 3 of centrifugation of the reconstituted vegetable juice is
- the duration of centrifugation step 3) can vary from approximately 15 to 25 minutes.
- Centrifugation step 3 is preferably carried out at a
- step 3 a centrifuged green juice is obtained which contains very few particles and a centrifugation pellet which can itself also be upgraded, for example, by methanization or be used for animal feed.
- step 3) of centrifugation makes it possible to eliminate the fine particles present in suspension in the reconstituted liquid vegetable juice which will then be used in step 4) of chromatographic retention.
- the method may further comprise, before performing step 1) of
- a step of prior centrifugation of the vegetable juice resulting from the pressing of a vegetable biomass containing Rubisco is preferably carried out under the same conditions as those defined above for step 3) to lead to a previously centrifuged juice which is then used in step 1) of
- chromatographic retention step 4 comprises, in this order, the following sub-steps:
- Sub-step 4a) allows the preparation of the ion exchange resin before its use in the chromatography column.
- the buffer solution used during this sub-step 4a) is preferably a phosphate buffer solution having a pH of approximately 6.0 ⁇ 0.2.
- the resin is preferably equilibrated with said buffer used for the washes.
- the buffer serving for equilibration can for example be introduced into the chromatography column with a feed rate of approximately 10 mL / min.
- the duration of the sub-step 4a) of rinsing and equilibration of the resin is preferably at least one hour.
- the purpose of the fixing sub-step 4b) (also called the loading step) is to fix the molecule of interest, here Rubisco, on the resin. This step also results in the binding of other molecules present in the centrifuged vegetable juice (in particular of proteins other than Rubisco, of polysaccharides, of polyphenols, etc.).
- Sub-step 4b) is preferably carried out at a pH of between 6.0 and 8.0 ⁇ 0.2, and even more preferably between 6.0 and 7.5 ⁇ 0.2.
- the supply of the column with the centrifuged vegetable juice can be carried out in ascending or descending mode.
- the flow can be controlled, for example using a peristaltic pump or a diaphragm pump.
- the feed rate can vary from approximately 80 mL / min to approximately 200 mL / min.
- the superficial feed rate preferably varies from 0.2 cm / min to approximately 2 cm / min.
- Sub-step 4b) leads to the recovery of a vegetable juice depleted in Rubisco, which is eliminated or upgraded according to its protein content, the Rubisco remaining fixed on the ion exchange resin.
- the washing sub-step 4c) makes it possible to remove from the column the remains of the centrifuged vegetable juice.
- the elution step 5 using a saline solution, makes it possible to recover the Rubisco fixed on the resin, then, once the Rubisco is recovered, to clean the resin for a new later use.
- This elution step 5) is preferably carried out using a saline solution (ion Na, K, Br, sulfate, Mg, etc.), and in particular a solution of NaCl, for example at 0.5 mol / L, which allows all of the Rubisco fixed on the resin to be eluted.
- a saline solution ion Na, K, Br, sulfate, Mg, etc.
- NaCl for example at 0.5 mol / L, which allows all of the Rubisco fixed on the resin to be eluted.
- isoelectric of the Rubisco can also be used.
- a gradient or steps of concentration can also be used.
- the surface speed of the saline solution preferably varies from 0.1 to 1.5 cm / min approximately and even more
- conditioning step 6) makes it possible to lower the salt content of the Rubisco saline solution obtained at the end of elution step 5) and, if necessary, to modify the nature of the salt (by example replacing sodium with potassium in order to make it more compatible with certain uses such as in the case of a diet low in sodium).
- Stage 6) of conditioning also allows post-purification of the solution. Indeed, at the end of this step, a saline solution (low salinity: of the order of 20 to 100 mM) of purified Rubisco is obtained, as well as a saline solution containing impurities.
- conditioning step 6) can be carried out by size exclusion chromatography or by ultrafiltration in mode.
- step 6) of conditioning is carried out by ultrafiltration in diafiltration mode using a replacement solution for the elution solution (for example a 50 mM or 100 mM phosphate buffer).
- a replacement solution for the elution solution for example a 50 mM or 100 mM phosphate buffer.
- This ultrafiltration technique is preferred to size exclusion chromatography because it is easy to transpose to an industrial scale and also makes it possible to concentrate the purified Rubisco solution.
- Ultrafiltration in diafiltration mode can in particular be carried out in stirred cells (Amicon® for example) using a polyethersulfone (PES) membrane having a cutting diameter of 300 kDa and at a working pressure of 0.5 to 1, Approx. 1 bar, or using tangential filtration cartridges.
- the purified Rubisco solution has a salt concentration of 3 to 7 g / L, and even more preferably of approximately 4 to 6 g / L. This solution can be used directly or it can be stored.
- the method according to the present invention then preferably comprises an additional step 7) of dehydration of said solution.
- This dehydration step 7) is preferably carried out by spray drying at an outlet temperature ranging from 65 to 75 ° C. This step 7) can therefore be carried out under the same conditions as those applied during step 1 and as described above.
- a purified Rubisco powder is then obtained, with a water content between 4 and 10% by mass, preferably between 4% and 6%, which can be stored more easily.
- step 1) of spray-drying the vegetable juice makes it possible to obtain a dried vegetable juice which keeps very well over a long period, which makes it possible to make the flow of vegetable juice usable in the process independent from that of biomass , the latter being seasonal and highly variable, in particular because of climatic conditions,
- animal feed production channels for example concentrated extracts of alfalfa
- the process (materials and chemicals) can be compatible with the food, cosmetic or pharmaceutical fields.
- FIG. 1 shows photos of fresh alfalfa juice (Figure la) and spray dried alfalfa juice ( Figure lb).
- FIG. 2 shows a diagram of a separation device
- FIG. 3 represents the elution curve of Rubisco isolated in step 5) of example 1.
- concentration of Rubisco in the fraction collected (in g / L) depends on the elution volume (in L).
- t R is the retention time of the protein on the column, characteristic of the affinity of the protein by the support.
- FIG. 4 represents a diagram of the ultrafiltration device used during step 6) for conditioning the Rubisco saline solution prepared in Example 1.
- FIG. 5 represents the chromatograms obtained by HPCL-SEC of the initial Rubisco solution before ultrafiltration, of the first sample of the permeate and of the retentate obtained at the end of the ultrafiltration for the experiment of step 6) of Example 1 .
- FIG. 6 represents the elution curves of Rubisco on a column containing a Q Sepharose ® Fast Flow resin (FIG. 6a), a column containing a Capto Q Impress ® resin (figure 6b) or a column containing an ANX Sepharose ® 4 Fast Flow resin (figure 6c).
- a purified Rubisco solution was prepared from a plant biomass derived from a dark green spring alfalfa with short, thin stems, without flowers, and a high leaf / stem ratio.
- alfalfa vegetable juice was prepared as follows.
- the fresh alfalfa (leaves and stems) was pre-crushed using an electric grinder with an automatic drive cutting system (AXT 25 TC, Bosch) in order to reduce the size of the stems to a length of approximately 5 to 10 cm.
- AXT 25 TC automatic drive cutting system
- the pre-crushed alfalfa was then pressed using a screw press manufactured by FEMAG Industries, with a maximum capacity of 100 kg / h.
- the press is made up of three main bodies: the feed hopper, the screw, and the outlet fitted with a shutter.
- An electrical box is used to adjust the operating parameters and a container is used to collect the juices produced.
- the set is made of stainless steel.
- the adjustable operating frequency (15 to 60 Hz) determines the speed of rotation of the screw.
- the pre-crushed alfalfa was weighed (79.6 kg) and then introduced into the press through the feed hopper.
- the frequency of rotation of the screw was set at 25 Hz (ie 12 revolutions / min.).
- the alfalfa vegetable juice obtained above was then dried by a spray drying process using a Niro Minor SD2 spray dryer from the company GEA with a capacity of 4 L / h at a speed of treatment of 1 L / h.
- the atomization conditions were set to have an inlet temperature of 120 ° C and an outlet temperature of 70 ⁇ 1 ° C.
- the dried alfalfa vegetable juice (Figure la) (powder with a residual water content of 6% by mass, Figure lb) was stored at -20 ° C.
- the powder of dehydrated alfalfa vegetable juice obtained above in the previous step was used to obtain a reconstituted alfalfa vegetable juice by adding a sodium carbonate solution in order to adjust its pH to 7 , 5, respecting the dry matter content determined in the starting juice, ie 7.2 ⁇ 0.3%.
- the reconstituted alfalfa vegetable juice obtained above in the previous step was then centrifuged at a speed of 6000 revolutions / min for 20 minutes at a temperature of 4 ° C.
- the centrifuged juice thus obtained had a pH of 7.5.
- the Rubisco concentration of the juice thus obtained was determined by HPLC SEC, with a Shodex KW-804 column, exclusion limit 10 6 Da and UV-Vis detector at 280 nm.
- the mobile phase is a phosphate buffer solution at pH 6.8 and at a flow rate of 1 mL / min. Data acquisition as well that the results are processed by the UNICORN 5.1® software (Cityva).
- the Rubisco concentration of this juice was 4.1 g / L.
- the conductivity of the juice was 12.1 mS / cm.
- the centrifuged juice thus obtained was then used in the next chromatographic retention step.
- the chromatographic retention was carried out using the ANX Sepharose® 4 Fast Flow resin manufactured by the company Cityva on the chromatographic separation device shown in FIG. 2 attached.
- This device 1 comprises a chromatography column 2 sold under the trade name Resolute FM 40 by the company Pal I, with an internal diameter of 140 mm containing the resin 3 and said column 2 being equipped with an adapter (not shown) allowing to adjust the height of the bed of resin 3 during the filling of column 2.
- Column 2 is supplied in ascending (or possibly descending) mode using pumps 4 and 4 '(membrane pump 4 for the feeding of the reconstituted vegetable juice centrifuged and peristaltic pump 4 'for feeding other solutions).
- the device 1 also comprises a first reservoir 5 containing
- Valves 9 and 9 ' are arranged downstream of the two reservoirs 5 and 7 and upstream of the pumps 4 and 4'.
- the valves 9 and 9 ' make it possible to successively introduce into column 2 the equilibration solution 6a, the centrifuged vegetable juice to be treated 8, the washing water 6b and the eluting saline solution 6c.
- Valves 10 and 10 'located respectively downstream of pumps 4 and 4' and upstream of column 2 make it possible, if necessary, to isolate column 2. At the outlet of column 2, the fractions are collected at the using a fraction collector 11.
- Table 1 summarizes the main characteristics of the fixed bed, the operating conditions and the parameters of the chromatographic separation device 1 used.
- column 2 was washed with demineralized water, until the conductivity of the water at the outlet of the column 2 is very weak and stable.
- a saline solution of elution 6c consisting of a sodium chloride solution of concentration 0.5 mol / L, was used to proceed with the elution of the Rubisco fixed on the resin 3.
- the elution step was carried out using 6000 mL of this sodium chloride solution, and with a flow rate of 32.3 mL / min, which corresponds to a surface speed of 0.21 cm / min.
- the total elution time was 186 minutes.
- the Rubisco elution curve is given in Figure 3 attached.
- the concentration of Rubisco in the fraction collected (in g / L) is a function of the elution volume (in L).
- the concentration of Rubisco in the eluting solution is determined by HPLC SEC.
- t R is the chromatographic retention time.
- the conditioning of the Rubisco saline solution obtained above in the previous step was carried out by ultrafiltration in diafiltration mode using an Amicon ® 8400 stirred cell (Merck-Millipore) with a volume of 400 mL , said cell being equipped with a polyethersulfone (PES) membrane, compatible with the production of drinking water, with a cutting diameter of 300 kD (cutoff threshold), said membrane being sold under the reference PBMK by the company Merck-Millipore.
- the area of the membrane was 41.8 cm 2 .
- the operating mode of Amicon® cells is a frontal flow mode (perpendicular to the membrane), which thanks to the
- agitation mechanism seeks to approximate the conditions of a
- This device 12 comprises a pressurized tank 13 with a capacity of 5 L (Sartorius, France) containing the solution of
- a valve 16 makes it possible to adjust the pressure of the compressed air 15, the latter being measured using a manometer 17 located downstream of the valve 16.
- the mass of the permeate 18 at the outlet of the cell 14 is measured over time and monitored by an electronic balance 19 (Sartorius 1500S, France), connected to a data acquisition system 20.
- the filtration cell 14 is composed of a support 14a which maintains the membrane (not
- the Rubisco saline solution was pre-filtered using a 0.45 ⁇ m filter.
- the system was filled with water ultra-pure thanks to the pressurized reservoir 13, and a high pressure (at least 0.1 bar in addition to the transmembrane pressure which will be applied) was maintained for 10 minutes in order to compact the membrane and to properly moisten the pores.
- the initial Rubisco content of the saline solution was first measured by size exclusion chromatography (HPLC-SEC) using an ATKA Purifier high pressure chromatograph (Cityva), with a KW- column. 804 (Shodex) and Unicom 5.1 data acquisition software.
- the initial Rubisco content of the saline solution was 16.5 g / L.
- the filtration cell 14 was completely filled with the Rubisco saline solution obtained above in the previous step.
- a volume of 4 L of 50 mmol / L phosphate buffer solution at pH 7.5 was placed in the pressurized tank 13. Once the assembly was carried out, the filtration cell 14 was stirred at 1200 revolutions / min and a 0.7 bar pressure was applied to the filtration device using the valve 16 making it possible to adjust the pressure of the compressed air 15. This pressure was kept constant throughout the ultrafiltration step.
- the mass of permeate 18 recovered was monitored by the data acquisition system 20 throughout the duration of the ultrafiltration.
- the concentration of purified Rubisco in the retentate was 15.4 g / L, which corresponds to a very low loss rate of 6.4%.
- the absorbance at 280 nm is a function of the volume (mL).
- the curve in long and broken lines (highest curve)
- the curve in short and broken lines corresponds to the first sample of the permeate and the curve in solid line corresponds to the retentate.
- the ultrafiltration step makes it possible not only to condition the Rubisco saline solution obtained above in the previous step with the aim of replacing NaCl with a phosphate buffer, but also to carry out a post-purification of Rubisco (disappearance of the peaks located between 11 and 15 mL on the chromatograph corresponding to the retentate compared to the two other curves corresponding respectively to the initial Rubisco solution before ultrafiltration and to the first sample of the permeate.
- a solution of Rubisco purified from a plant biomass obtained from alfalfa was also prepared according to the various stages as detailed above in Example 1, but replacing the ANX Sepharose® 4 Fast resin.
- Flow used in Example 1 by the anionic resins Q Sepharose® Fast Flow and Capto Q Impress® sold by the company Cityva, which can also be used according to the process of the invention.
- the column used was an XK type column 16/20 having an internal diameter of 16 mm (Cityva).
- the preparation was also carried out using the ANX Sepharose® 4 Fast Flow resin used in Example 1.
- Co is the initial concentration of Rubisco in the centrifuged reconstituted alfalfa juice.
- Example 3 Charging the Rubisco with ion exchanging resins of various matrices
- the column used was an XK 16/20 type column having an internal diameter of 16 mm (Cityva).
- Co is the initial concentration of Rubisco in the centrifuged alfalfa juice.
- Example 4 Dehydration step 1): Atomization of alfalfa water at different inlet temperatures of the atomizer
- the juices were stored as they were produced in a refrigerated tank at 4 ° C until the end of production. The juices were then transferred into flasks and then stored at 4 ° C. until the moment of drying (between 1 and 3 days). As can be seen in Table 9 above, the two juices produced had very similar characteristics, knowing that there is a natural variability linked to the raw material. Spray drying tests were then carried out.
- the equipment used for the tests is a Niro Minor SD2 atomiser-dryer from GEA with a capacity of 4 L / h.
- second temperature of step 1 of dehydration.
- the obtained powder was stored at -20 ° C.
- the powder obtained after drying was reconstituted in ultra pure water, at a dry matter content value equal to that of the juice before atomization (see Table 9).
- the reconstituted juice was then centrifuged at 6000 rpm. at 4 ° C for 20 min. and analyzed by HPLC-SEC at 280 nm, under the same conditions as those of Example 1, in order to determine its Rubisco concentration.
- the effect of atomization temperature on the concentration of Rubisco in the reconstituted juice was studied.
- Table 11 below presents a summary of the results obtained by material balance in terms of mass of juice and mass of Rubisco obtained after atomization, relative to the quantities which were treated.
- inlet temperature of the atomizer is less than 200 ° C, and even more preferably less than 140 ° C. a very particularly suitable temperature is 120 ° C as in Examples 1 and 2 above.
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR1907012A FR3097723B1 (en) | 2019-06-27 | 2019-06-27 | RUBISCO PRODUCTION PROCESS FROM VEGETABLE BIOMASS |
PCT/FR2020/051110 WO2020260831A1 (en) | 2019-06-27 | 2020-06-25 | Process for the production of rubisco from a plant biomass |
Publications (1)
Publication Number | Publication Date |
---|---|
EP3989731A1 true EP3989731A1 (en) | 2022-05-04 |
Family
ID=68343051
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP20747040.2A Pending EP3989731A1 (en) | 2019-06-27 | 2020-06-25 | Process for the production of rubisco from a plant biomass |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP3989731A1 (en) |
FR (1) | FR3097723B1 (en) |
WO (1) | WO2020260831A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
PT117122A (en) * | 2021-03-17 | 2022-11-30 | Inst Superior Agronomia | PROCESS FOR OBTAINING PURE RUBISCO FROM A PHOTOSYNTHETIC MATERIAL |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2819685B1 (en) | 2001-01-23 | 2005-02-25 | Viridis | PROCESS FOR TREATING GREEN JUICE FROM PRESSING OF PROTEIN-RICH FOLIAR MATERIAL SUCH AS LUZERNE |
WO2011078671A1 (en) | 2009-12-22 | 2011-06-30 | Nizo Food Research B.V. | Process for isolating a dechlorophylllized rubisco preparation from a plant material |
FR2974817B1 (en) | 2011-05-04 | 2017-06-23 | Valagro Carbone Renouvelable Poitou-Charentes | PROCESS FOR OBTAINING PROTEIN EXTRACT FROM LUZERNE AND VALORISABLE CO-PRODUCTS |
-
2019
- 2019-06-27 FR FR1907012A patent/FR3097723B1/en active Active
-
2020
- 2020-06-25 WO PCT/FR2020/051110 patent/WO2020260831A1/en active Application Filing
- 2020-06-25 EP EP20747040.2A patent/EP3989731A1/en active Pending
Also Published As
Publication number | Publication date |
---|---|
FR3097723B1 (en) | 2021-06-18 |
WO2020260831A1 (en) | 2020-12-30 |
FR3097723A1 (en) | 2021-01-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP3300610B1 (en) | Method for fractioning pea solubles | |
EP1400537B1 (en) | Extraction of the components from pea flour | |
JP2016005455A (en) | Zein composition | |
CN105324040A (en) | Tobacco-derived protein compositions | |
EP2414379B1 (en) | Method for obtaining a preparation of beta-amylases from the soluble fractions of starch plants | |
Marson et al. | Serial fractionation of spent brewer's yeast protein hydrolysate by ultrafiltration: A peptide-rich product with low RNA content | |
EP3989731A1 (en) | Process for the production of rubisco from a plant biomass | |
CN105924498B (en) | Preparation method of mung bean protein with blood fat reducing effect | |
EP1297116B1 (en) | Method for isolating and purifying a protein | |
CN107594541A (en) | A kind of the membrane separation and purification technology technique for preparing bitter gourd polypeptide albumen, bitter gourd polypeptide protein extract and its application | |
EP4045542B1 (en) | Soluble corn steep liquor | |
EP2920160B1 (en) | Method for obtaining polyphenols from a vegetable raw material containing same | |
EP2451825A1 (en) | Use of a co-product from a method for extracting lysozyme from egg whites, in order to obtain at least one basic egg white protein | |
EP3233076A1 (en) | Process for obtaining a composition enriched with dihydroquercetin or with tannins | |
WO2024088564A1 (en) | Clear, water-soluble plant fraction | |
EP3924462A1 (en) | Method for obtaining an aqueous solution enriched with blue pigment | |
EP3013948B1 (en) | Method for extracting beta-amylases from a soluble fraction of a starch plant and in the presence of a pectinase | |
JP5927833B2 (en) | Method for producing a polyamine composition from a plant | |
JP2009183276A (en) | Rice bran leachate having enzyme activity, product obtained from the same rice bran leachate, and method for producing the same | |
BE577712A (en) |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: UNKNOWN |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE |
|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE |
|
17P | Request for examination filed |
Effective date: 20211227 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
DAV | Request for validation of the european patent (deleted) | ||
DAX | Request for extension of the european patent (deleted) | ||
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
17Q | First examination report despatched |
Effective date: 20240516 |