CN110229803A - Stabilized enzyme formulations and its preparation process and application containing alkalinity or neutral proteinase - Google Patents
Stabilized enzyme formulations and its preparation process and application containing alkalinity or neutral proteinase Download PDFInfo
- Publication number
- CN110229803A CN110229803A CN201910481004.9A CN201910481004A CN110229803A CN 110229803 A CN110229803 A CN 110229803A CN 201910481004 A CN201910481004 A CN 201910481004A CN 110229803 A CN110229803 A CN 110229803A
- Authority
- CN
- China
- Prior art keywords
- alkalinity
- compound stabilizer
- neutral
- enzyme solution
- mercapto
- 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
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 181
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 181
- 239000000203 mixture Substances 0.000 title claims abstract description 76
- 238000009472 formulation Methods 0.000 title claims abstract description 47
- 238000002360 preparation method Methods 0.000 title claims abstract description 46
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 title claims abstract description 38
- 239000002131 composite material Substances 0.000 claims abstract description 129
- 108091005804 Peptidases Proteins 0.000 claims abstract description 117
- 239000004365 Protease Substances 0.000 claims abstract description 116
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 116
- 235000019419 proteases Nutrition 0.000 claims abstract description 115
- 150000001875 compounds Chemical class 0.000 claims abstract description 111
- 239000003381 stabilizer Substances 0.000 claims abstract description 106
- 238000000746 purification Methods 0.000 claims abstract description 74
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 73
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 71
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 70
- 239000003223 protective agent Substances 0.000 claims abstract description 66
- 238000005189 flocculation Methods 0.000 claims abstract description 64
- 230000016615 flocculation Effects 0.000 claims abstract description 64
- 230000007935 neutral effect Effects 0.000 claims abstract description 51
- 239000007788 liquid Substances 0.000 claims abstract description 49
- 238000000034 method Methods 0.000 claims abstract description 48
- 238000000855 fermentation Methods 0.000 claims abstract description 38
- 230000004151 fermentation Effects 0.000 claims abstract description 38
- 150000003839 salts Chemical class 0.000 claims abstract description 37
- 229920000592 inorganic polymer Polymers 0.000 claims abstract description 36
- 229920000620 organic polymer Polymers 0.000 claims abstract description 36
- 238000001914 filtration Methods 0.000 claims abstract description 27
- 238000007906 compression Methods 0.000 claims abstract description 19
- 230000006835 compression Effects 0.000 claims abstract description 19
- 238000000967 suction filtration Methods 0.000 claims abstract description 14
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 13
- 239000003599 detergent Substances 0.000 claims abstract description 8
- 238000005342 ion exchange Methods 0.000 claims abstract description 8
- 238000007670 refining Methods 0.000 claims abstract description 7
- 235000006708 antioxidants Nutrition 0.000 claims description 69
- 238000002156 mixing Methods 0.000 claims description 57
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 claims description 50
- 235000002639 sodium chloride Nutrition 0.000 claims description 41
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 36
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims description 28
- 239000008394 flocculating agent Substances 0.000 claims description 26
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 24
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 claims description 22
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 18
- 229920002401 polyacrylamide Polymers 0.000 claims description 16
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 14
- 108010024636 Glutathione Proteins 0.000 claims description 14
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 14
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 14
- 235000018417 cysteine Nutrition 0.000 claims description 14
- 150000002016 disaccharides Chemical class 0.000 claims description 14
- 229960003180 glutathione Drugs 0.000 claims description 14
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 claims description 14
- 235000019345 sodium thiosulphate Nutrition 0.000 claims description 14
- 150000005846 sugar alcohols Polymers 0.000 claims description 14
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 13
- 229930003268 Vitamin C Natural products 0.000 claims description 13
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 claims description 13
- 235000019154 vitamin C Nutrition 0.000 claims description 13
- 239000011718 vitamin C Substances 0.000 claims description 13
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 235000011187 glycerol Nutrition 0.000 claims description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 8
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 claims description 8
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 7
- 229930006000 Sucrose Natural products 0.000 claims description 7
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 7
- 239000001110 calcium chloride Substances 0.000 claims description 7
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 7
- RUTXIHLAWFEWGM-UHFFFAOYSA-H iron(3+) sulfate Chemical compound [Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O RUTXIHLAWFEWGM-UHFFFAOYSA-H 0.000 claims description 7
- 229910000360 iron(III) sulfate Inorganic materials 0.000 claims description 7
- 239000005720 sucrose Substances 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 6
- 244000063299 Bacillus subtilis Species 0.000 claims description 5
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 5
- 101150103639 PB1 gene Proteins 0.000 claims description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 5
- 239000005864 Sulphur Substances 0.000 claims description 5
- 235000010323 ascorbic acid Nutrition 0.000 claims description 5
- 239000011668 ascorbic acid Substances 0.000 claims description 5
- 229960005070 ascorbic acid Drugs 0.000 claims description 5
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 241001507822 Aspergillus terricola Species 0.000 claims description 4
- 241000193422 Bacillus lentus Species 0.000 claims description 4
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 4
- SZXQTJUDPRGNJN-UHFFFAOYSA-N dipropylene glycol Chemical compound OCCCOCCCO SZXQTJUDPRGNJN-UHFFFAOYSA-N 0.000 claims description 4
- VHJLVAABSRFDPM-ZXZARUISSA-N dithioerythritol Chemical compound SC[C@H](O)[C@H](O)CS VHJLVAABSRFDPM-ZXZARUISSA-N 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 4
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 4
- 235000011152 sodium sulphate Nutrition 0.000 claims description 4
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 claims description 3
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 3
- XSTXAVWGXDQKEL-UHFFFAOYSA-N Trichloroethylene Chemical compound ClC=C(Cl)Cl XSTXAVWGXDQKEL-UHFFFAOYSA-N 0.000 claims description 3
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 3
- 241000186046 Actinomyces Species 0.000 claims description 2
- 241000194108 Bacillus licheniformis Species 0.000 claims description 2
- UNXHWFMMPAWVPI-QWWZWVQMSA-N D-threitol Chemical compound OC[C@@H](O)[C@H](O)CO UNXHWFMMPAWVPI-QWWZWVQMSA-N 0.000 claims description 2
- 150000001408 amides Chemical class 0.000 claims description 2
- QQONPFPTGQHPMA-UHFFFAOYSA-N propylene Natural products CC=C QQONPFPTGQHPMA-UHFFFAOYSA-N 0.000 claims description 2
- 125000004805 propylene group Chemical group [H]C([H])([H])C([H])([*:1])C([H])([H])[*:2] 0.000 claims description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims 1
- 229920002472 Starch Polymers 0.000 claims 1
- 230000003064 anti-oxidating effect Effects 0.000 claims 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 claims 1
- 239000011777 magnesium Substances 0.000 claims 1
- 229910052749 magnesium Inorganic materials 0.000 claims 1
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical group CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims 1
- 229920000642 polymer Polymers 0.000 claims 1
- 238000003825 pressing Methods 0.000 claims 1
- 239000008107 starch Substances 0.000 claims 1
- 235000019698 starch Nutrition 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 16
- 238000005119 centrifugation Methods 0.000 abstract description 2
- 238000004321 preservation Methods 0.000 abstract description 2
- 239000003513 alkali Substances 0.000 description 68
- 239000012530 fluid Substances 0.000 description 31
- 230000008569 process Effects 0.000 description 14
- 238000004140 cleaning Methods 0.000 description 11
- 238000012545 processing Methods 0.000 description 11
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 10
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 9
- 239000004411 aluminium Substances 0.000 description 9
- 229910052782 aluminium Inorganic materials 0.000 description 9
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 9
- 230000006641 stabilisation Effects 0.000 description 8
- 238000011105 stabilization Methods 0.000 description 8
- 238000005406 washing Methods 0.000 description 8
- 125000000647 trehalose group Chemical group 0.000 description 7
- 239000004094 surface-active agent Substances 0.000 description 6
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 5
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 3
- 241000235058 Komagataella pastoris Species 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 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
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 238000000703 high-speed centrifugation Methods 0.000 description 2
- 125000001165 hydrophobic group Chemical group 0.000 description 2
- 229910017053 inorganic salt Inorganic materials 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 125000000185 sucrose group Chemical group 0.000 description 2
- 125000003396 thiol group Chemical group [H]S* 0.000 description 2
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 108010051873 alkaline protease Proteins 0.000 description 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 1
- 230000003281 allosteric effect Effects 0.000 description 1
- 235000010210 aluminium Nutrition 0.000 description 1
- 238000012870 ammonium sulfate precipitation Methods 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000012869 ethanol precipitation Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000010200 folin Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000003262 industrial enzyme Substances 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- -1 thio sulphur Chemical compound 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D9/00—Compositions of detergents based essentially on soap
- C11D9/04—Compositions of detergents based essentially on soap containing compounding ingredients other than soaps
- C11D9/22—Organic compounds, e.g. vitamins
- C11D9/40—Proteins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
-
- 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
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
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)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cosmetics (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a kind of stabilized enzyme formulations and its preparation process and application containing alkalinity or neutral proteinase.The stabilized enzyme formulations are to be made by the following method with technique.Alkalinity or natural protease thick fermentation liquid are subjected to flocculation treatment using the composite flocculation agent that organic/inorganic polymer and inorganic salts are formulated, limpid alkalinity or neutral protein enzyme solution are obtained by filtering, suction filtration or plate compression, compound stabilizer is added thereto again, is refining to obtain the alkalinity of the purification containing compound stabilizer or neutral protein enzyme solution through ultrafiltration or excessively ion exchange column；Then alkalinity or neutral protein enzyme solution and composite antioxidant will be refined containing compound stabilizer and mercapto-protective agent is hybridly prepared into the stabilized enzyme formulations containing alkalinity or neutral proteinase.The stabilized enzyme formulations are applied in detergent product.The present invention saves the centrifugation step of highly energy-consuming, reaches energy-efficient target, and compound stabilizer, composite antioxidant and mercapto-protective agent, which is added, can enable enzyme activity obtain long-acting preservation.
Description
Technical field
The present invention relates to cleaning product technical fields, and in particular to a kind of stabilized enzyme formulations containing alkalinity or neutral proteinase
And its preparation process and application.
Background technique
Alkali protease is very important a kind of industrial enzymes, is widely used in the industries such as food, washing, leather,
Especially in washing industry.In daily life, clothes washing is can not avoid the problem of, although with society into
Step, it is already possible to replace manpower to carry out clothes washing using machine, but the detergent selection in washing process is still a asks
Topic.Main component in detergent is surfactant, and surfactant has hydrophilic group and hydrophobic group simultaneously, in washing process
In, the spot on multiple surfactant molecule package clothings, hydrophobic group is in conjunction with spot, and hydrophilic group and water molecules,
To form a micella group, using mechanical friction campaign, micella group is removed from clothing, achievees the effect that washing.But
It is that there are a deficiencies during this, that is, surfactant easily can form micella group with small-molecule substance,
But have for the spot power of macromolecular and do not capture, need to add more surfactants, this will certainly cause environment worse
Influence.After protease is added in detergent, the macromolecular components in many spots can be decomposed into small molecule, be conducive to table
The effect of face activating agent, improves detergency, reduces the dosage of surfactant, reduce environmental pollution.
Enzyme does not pollute in the natural environment as a kind of natural biological agent, and can natural degradation, for ring
Border is not polluted, and compared with traditional detergent, enzyme detergent has big advantage, is to wash the important development side of industry
To.
The source of present most of biological enzyme is all by fungi (Pichia pastoris) or bacterium (bacillus subtilis)
What fermentation generated.After fermentation, since target product is present in fermentation liquid, and the content of target product in fermentation liquor
It is relatively low, it is therefore desirable to separating-purifying to be carried out to fermentation liquid, to obtain the target product of high-purity.Currently, mainly passing through
The methods of high speed centrifugation, salt fractionation, classification alcohol precipitation realize isolating and purifying for fermentation liquid, extract target product, rough processing
Program is as follows:
Fermentation liquid → high speed centrifugation → ammonium sulfate precipitation (salt fractionation) → dialysis desalting → ethanol precipitation (point
Grade alcohol precipitation) → ion-exchange chromatography.
Although the target product of the available high-purity of isolation and purification method of current mainstream, due to isolating and purifying
Journey does not take into account that the otherness of different fermentations liquid, is purified with same process, and purification process will cause a large amount of energy
Loss will also result in the loss of a large amount of target product, and Simultaneous purification technique is also relatively long, and it is too many to be related to program, purification efficiency
It is relatively low, and the phase transition process of target product is had in purification process.
Therefore the method for finding a kind of separation of fermentative broth purifying of rapidly and efficiently low consumption, to promotion alkali protease liquid system
The industrialization of agent is of great significance, this will greatly improve the production technology level of China's micro-organism enzyme preparation, pushes me
The development of state's Enzymes Industry, promotes the further development of washing industry.
In addition, sulfydryl is very important for most of enzymes, these sulfydryls may participate in catalysis and substrate knot
Close, form allosteric site, the three or four structure for maintaining enzyme etc., therefore, so that sulfydryl is kept native state is to keep many enzymes steady
Surely one of essential condition changed.
Summary of the invention
For energy loss existing for solution alkalinity or neutral proteinase fermentation liquid subtractive process, big, target product damages the present invention
Lose, the purifying process time it is long, be related to that program is more, sulfhydryl protected problem in the low problem of purification efficiency and protease proposes one
Stabilized enzyme formulations and its technique and application of the kind containing alkalinity and neutral proteinase.
In order to solve the above technical problems, the invention adopts the following technical scheme:
A kind of stabilized enzyme formulations containing alkalinity or neutral proteinase, the stabilized enzyme formulations are obtained by the following method；
Specially alkalinity or natural protease thick fermentation liquid are after composite flocculation agent flocculation treatment, by filtering, suction filtration or sheet frame pressure
Filter obtains limpid alkalinity or neutral protein enzyme solution, then compound stabilizer is added thereto and carries out ultrafiltration or crosses ion exchange column essence
System；Alkalinity or neutral protein enzyme solution and composite antioxidant and mercapto-protective agent after purification are configured to stabilized enzyme formulations.
Further, it is described alkalinity or natural protease thick fermentation liquid be by Bacillus bacteria or yeast fermenting and producing,
Middle Bacillus bacteria refers to bacillus amyloliquefaciens, bacillus lentus, bacillus subtilis or bacillus licheniformis；It is described compound
Flocculant is the mixture of organic/inorganic polymer and inorganic salts, the weight percent of component contained by the composite flocculation agent are as follows:
5%~98% organic/inorganic polymer flocculants, surplus are inorganic salts flocculant；The organic/inorganic polymer is propylene
Any one in amide PAM, PB1 gene, bodied ferric sulfate SPFS or polyaluminum ferric chloride PAFC, inorganic salts are
The mixing of one of calcium sulfate, calcium chloride, aluminum sulfate, sodium sulphate, sodium chloride, magnesium sulfate or several arbitrary proportions；It is described
The compound stabilizer of addition is the 0.1%~40% of limpid alkalinity or neutral proteinase fermentation liquid weight；The compound stabilizer
For the mixture that non-reducible disaccharide and polyalcohol form, the weight ratio of non-reducible disaccharide and polyalcohol is 1:1~10, wherein non-go back
Former disaccharides is one of sucrose, trehalose or the mixing of their arbitrary proportions, and polyalcohol is propylene glycol, dipropylene glycol, third
The mixing of one of triol or several arbitrary proportions；Purification alkalinity or neutral protein enzyme solution, composite antioxidant and
The weight percent of mercapto-protective agent are as follows: 35%~98% purification basic protein enzyme solution, 0.01%~5% compound anti-oxidation
Agent, surplus are mercapto-protective agent；The composite antioxidant is VitAVitE, in ascorbic acid, sodium thiosulfate
The mixing of one or several kinds of arbitrary proportions；The mercapto-protective agent is cysteine, glutathione, 2 mercapto ethanol, two sulphur
The mixing of one or more of threitol, dithioerythritol arbitrary proportion.
A kind of preparation process of the stabilized enzyme formulations containing alkalinity or neutral proteinase are as follows: using organic/inorganic polymer and
Alkalinity or natural protease thick fermentation liquid are carried out flocculation treatment by the composite flocculation agent that inorganic salts are formulated, by filtering, taking out
Filter or plate compression obtain limpid alkalinity or neutral protein enzyme solution, then compound stabilizer is added thereto, through ultrafiltration or mistake
Ion exchange column is refining to obtain the alkalinity of the purification containing compound stabilizer or neutral protein enzyme solution；Then it will contain compound stabilizer essence
Alkaline or neutral protein enzyme solution and composite antioxidant and mercapto-protective agent are hybridly prepared into containing alkaline or neutral proteinase
Stabilized enzyme formulations.
Further, it is described alkalinity or natural protease thick fermentation liquid be by Bacillus bacteria or yeast fermenting and producing,
Middle Bacillus bacteria refers to bacillus amyloliquefaciens, bacillus lentus, actinomyces, aspergillus terricola, bacillus subtilis or lichens
Bacillus.
Further, the weight percent of the composite flocculation agent and alkalinity or natural protease thick fermentation liquid component
Are as follows: 35%~99.5% alkalinity or natural protease thick fermentation liquid, 0.5%~65% composite flocculation agent.Preferred weight
Percentage are as follows: the alkalinity of 55%-99% or the composite flocculation agent of natural protease thick fermentation liquid, 1%-45%；Preferred weight
Measure percentage are as follows: the alkalinity of 75%-99% or the composite flocculation agent of natural protease thick fermentation liquid, 1%-25%；More preferably
Weight percent are as follows: the alkalinity of 80%-97% or the composite flocculation agent of natural protease thick fermentation liquid, 3%-20%.
Further, the composite flocculation agent is formulated by organic/inorganic polymer and inorganic salts, described compound
The weight percent of component contained by flocculant are as follows: 5%~98% organic/inorganic polymer flocculants, 2%~95% it is inorganic
Salt flocculant；The organic/inorganic polymer is polyacrylamide PAM, PB1 gene, bodied ferric sulfate SPFS or poly-
Close the mixing of one of aluminium chloride ferrum PAFC or several arbitrary proportions；The inorganic salts are calcium sulfate, calcium chloride, sulfuric acid
The mixing of one of aluminium, sodium sulphate, sodium chloride, magnesium sulfate or several arbitrary proportions.
Further, the compound stabilizer of the addition be it is limpid alkalinity or neutral protein enzyme solution weight 0.1%~
40%, the compound stabilizer is the mixture that non-reducible disaccharide and polyalcohol form, wherein non-reducible disaccharide and polyalcohol
Weight ratio is 1:1~10, and the non-reducible disaccharide is the mixing of one of sucrose, trehalose or several arbitrary proportions；Institute
State the mixing that polyalcohol is one of propylene glycol, dipropylene glycol, glycerine or several arbitrary proportions.
Further, it is described containing compound stabilizer purification alkalinity or neutral protein enzyme solution, composite antioxidant and
The weight percent of mercapto-protective agent are as follows: 35%~98% containing compound stabilizer purification alkalinity or neutral protein enzyme solution,
0.01%~5% composite antioxidant, surplus are mercapto-protective agent, purification alkalinity or neutral protein containing compound stabilizer
The sum of weight percent of enzyme solution, composite antioxidant and mercapto-protective agent is 100%.Preferred weight percent are as follows: 55%-
98% containing compound stabilizer refine 2709 basic protein enzyme solutions, the composite antioxidant of 0.1%-5%, surplus be sulfydryl protect
Protect agent；Preferred weight percent is 90%-98% containing compound stabilizer purification alkalinity or neutral protein enzyme solution, 1%-
3% composite antioxidant, surplus are mercapto-protective agent.
Further, the composite antioxidant is VitAVitE, vitamin C, ascorbic acid, thio sulphur
The mixing of one of sour sodium or several arbitrary proportions；The mercapto-protective agent is cysteine, glutathione, 2- sulfydryl
The mixing of one of ethyl alcohol, dithiothreitol (DTT), dithioerythritol or several arbitrary proportions.
The stabilized enzyme formulations containing alkalinity or neutral proteinase are applied in detergent product.
Compared with prior art, the invention has the following advantages:
Composite flocculation agent provided by the invention, can be by the solid impurity of 99% thallus, 95% or more in thick hair zymotic fluid
It is removed by flocculation, obtains very limpid alkalinity or neutral protein enzyme solution, the centrifugation step of highly energy-consuming can be saved, reach section
Can target, while after ultrafiltration, obtain purification basic protein enzyme solution mixed with composite antioxidant, mercapto-protective agent after
The stable protease preparation containing alkali protease is obtained by processing, enzyme activity can obtain long-acting preservation.
Specific embodiment
For the technical solution that the present invention is further explained, combined with specific embodiments below to technical solution of the present invention into
Row clearly and completely describes, it is clear that and described embodiment is only a part of the embodiments of the present invention, rather than whole
Embodiment.Based on the embodiments of the present invention, those of ordinary skill in the art are obtained without making creative work
The every other embodiment obtained, shall fall within the protection scope of the present invention.
Embodiment 1
A kind of preparation process of the stabilized enzyme formulations containing alkali protease are as follows:
A kind of alkali protease thick hair zymotic fluid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then thereto after addition compound stabilizer, limpid alkali protease is obtained by filtering, plate compression
Liquid, then limpid basic protein enzyme solution is sent into ultrafilter and carries out ultrafiltration, obtain the purification alkali protease containing compound stabilizer
Liquid finally adds composite antioxidant and mercapto-protective agent into the purification basic protein enzyme solution containing compound stabilizer, is contained
The stabilized enzyme formulations of alkali protease.
The alkali protease thick hair zymotic fluid is fermented by Bacillus licheniformis2709 to be generated；
The weight percent of the composite flocculation agent and alkali protease thick hair zymotic fluid component are as follows: 99.5% protease is thick
Fermentation liquid, 0.5% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 25% organic/inorganic polymer flocculants,
75% inorganic salts flocculant；
The organic/inorganic polymer flocculants are the mixing of polyacrylamide and aluminium polychloride, mixing ratio 1:5；
Inorganic salts are aluminum sulfate；
The weight percent of the compound stabilizer be basic protein enzyme solution 0.1%, compound stabilizer be trehalose and
The mixture of propylene glycol, glycerine, mixed proportion 1:4:4；
The weight percent of the purification basic protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Than are as follows: the 97% purification basic protein enzyme solution containing compound stabilizer, 0.01% composite antioxidant, surplus are sulfhydryl protected
Agent；The weight percent of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
Than the sum of be 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is the mixture of cysteine, glutathione, mixed proportion 1:1.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stability containing alkali protease
Enzyme preparation is applied in cleaning product.
Alkali protease is shown in Table 1 using the method enzyme activity comparison before and after the processing of the present embodiment.
Embodiment 2
A kind of preparation process of the stabilized enzyme formulations containing neutral proteinase are as follows:
A kind of natural protease thick fermentation liquid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then limpid alkali neutral proteinase is obtained by filtering, filtering after addition compound stabilizer thereto
Liquid, then limpid neutral protein enzyme solution is refining to obtain the purification neutral protein enzyme solution containing compound stabilizer by ion exchange column,
Composite antioxidant and mercapto-protective agent are finally added into the purification neutral protein enzyme solution containing compound stabilizer, are obtained containing neutrality
The stabilized enzyme formulations of protease.
The natural protease thick fermentation liquid is fermented by bacillus amyloliquefaciens to be generated；
The weight percent of the composite flocculation agent and natural protease thick fermentation liquid component are as follows: 80% protease thick hair
Zymotic fluid, 20% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 85% organic/inorganic polymer flocculants,
15% inorganic salts flocculant；
The organic/inorganic polymer flocculants are the mixing of polyacrylamide and aluminium polychloride, mixing ratio 1:3；
Inorganic salts are calcium sulfate；
The weight percent of the compound stabilizer be neutral protein enzyme solution 0.5%, compound stabilizer be trehalose and
The mixture of propylene glycol, glycerine, mixed proportion 1:3:3；
The weight hundred of the purification neutral protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 90% purification neutral protein enzyme solution containing compound stabilizer, 0.1% composite antioxidant, surplus are sulfydryl
Protective agent；The weight of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
The sum of percentage is 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is the mixture of cysteine, glutathione, mixed proportion 1:1.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stabilization enzyme containing neutral proteinase
Preparation is used in cleaning product.
Neutral proteinase is shown in Table 1 using the method enzyme activity comparison before and after the processing of this implementation.
Embodiment 3
A kind of preparation process of the stabilized enzyme formulations containing neutral proteinase are as follows:
A kind of natural protease thick fermentation liquid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then limpid alkali neutral proteinase is obtained by filtering, filtering after addition compound stabilizer thereto
Liquid, then limpid neutral protein enzyme solution is refining to obtain the purification neutral protein enzyme solution containing compound stabilizer by ion exchange column,
Composite antioxidant and mercapto-protective agent are finally added into the purification neutral protein enzyme solution containing compound stabilizer, are obtained containing neutrality
The stabilized enzyme formulations of protease.
The natural protease thick fermentation liquid is domestic 3942 neutral proteinase of aspergillus terricola fermentation；
The weight percent of the composite flocculation agent and natural protease thick fermentation liquid component are as follows: 97% protease thick hair
Zymotic fluid, 3% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 80% organic/inorganic polymer flocculants,
20% inorganic salts flocculant；
The organic/inorganic polymer flocculants are the mixing of polyacrylamide and aluminium polychloride, mixing ratio 1:7；
Inorganic salts are the mixing of aluminum sulfate and calcium sulfate, mixed proportion 1:1；；
The weight percent of the compound stabilizer is the 1% of neutral protein enzyme solution, and compound stabilizer is trehalose, sucrose
With the mixture of propylene glycol, glycerine, mixed proportion 1:1:4:5；
The weight hundred of the purification neutral protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 95% purification basic protein enzyme solution containing compound stabilizer, 1% composite antioxidant, surplus are that sulfydryl is protected
Protect agent；
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is the mixture of cysteine, glutathione, mixed proportion 1:1.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stabilization enzyme containing neutral proteinase
Preparation is applied in cleaning product.
Neutral proteinase is shown in Table 1 using the method enzyme activity comparison before and after the processing of the present embodiment.
Embodiment 4
A kind of preparation process of the stabilized enzyme formulations containing neutral proteinase are as follows:
A kind of natural protease thick fermentation liquid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then limpid alkali neutral proteinase is obtained by filtering, filtering after addition compound stabilizer thereto
Liquid, then limpid neutral protein enzyme solution is refining to obtain the purification neutral protein enzyme solution containing compound stabilizer by ion exchange column,
Composite antioxidant and mercapto-protective agent are finally added into the purification neutral protein enzyme solution containing compound stabilizer, are obtained containing neutrality
The stabilized enzyme formulations of protease.
The natural protease thick fermentation liquid is fermented by aspergillus terricola to be generated；
The weight percent of the composite flocculation agent and natural protease thick fermentation liquid component are as follows: 93% protease thick hair
Zymotic fluid, 7% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 15% organic/inorganic polymer flocculants,
85% inorganic salts flocculant；
The organic/inorganic polymer flocculants are aluminium polychloride；Inorganic salts are aluminum sulfate；
The weight percent of the compound stabilizer is the 10% of neutral protein enzyme solution, and compound stabilizer is trehalose and third
The mixture of glycol, glycerine, mixed proportion 1:2:4；
The weight hundred of the purification neutral protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 55% purification neutral protein enzyme solution containing compound stabilizer, 1% composite antioxidant, surplus are that sulfydryl is protected
Protect agent；The weight hundred of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
Dividing the sum of ratio is 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is cysteine.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stabilization enzyme containing neutral proteinase
Preparation is applied in cleaning product.
Neutral proteinase is shown in Table 1 using the method enzyme activity comparison before and after the processing of the present embodiment.
Embodiment 5
A kind of preparation process of the stabilized enzyme formulations containing alkali protease are as follows:
A kind of alkali protease thick hair zymotic fluid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then thereto after addition compound stabilizer, limpid alkali protease is obtained by filtering, plate compression
Liquid, then limpid basic protein enzyme solution is sent into ultrafilter and carries out ultrafiltration, obtain the purification alkali protease containing compound stabilizer
Liquid finally adds composite antioxidant and mercapto-protective agent into the purification basic protein enzyme solution containing compound stabilizer, is contained
The stabilized enzyme formulations of alkali protease.
The alkali protease thick hair zymotic fluid is fermented by Bacillus licheniformis2709 to be generated；
The weight percent of the composite flocculation agent and alkali protease thick hair zymotic fluid component are as follows: 99% protease thick hair
Zymotic fluid, 1% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 90% organic/inorganic polymer flocculants,
10% inorganic salts flocculant；
The organic/inorganic polymer flocculants are the mixing of polyacrylamide, aluminium polychloride, bodied ferric sulfate, are mixed
Composition and division in a proportion is 1:5:1；Inorganic salts are aluminum sulfate；
The weight percent of the compound stabilizer is the 40% of basic protein enzyme solution, and compound stabilizer is sucrose and the third two
The mixture of alcohol, mixed proportion 1:4；
The weight hundred of the purification basic protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 35% purification basic protein enzyme solution containing compound stabilizer, 5% composite antioxidant, surplus are that sulfydryl is protected
Protect agent；The weight hundred of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
Dividing the sum of ratio is 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is the mixture of cysteine, glutathione, mixed proportion 1:3.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stabilization enzyme containing alkali protease
Preparation is applied in cleaning product.
Alkali protease is shown in Table 1 using the method enzyme activity comparison before and after the processing of the present embodiment.
Embodiment 6
A kind of preparation process of the stabilized enzyme formulations containing alkali protease are as follows:
A kind of alkali protease thick hair zymotic fluid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then thereto after addition compound stabilizer, limpid alkali protease is obtained by filtering, plate compression
Liquid, then limpid basic protein enzyme solution is sent into ultrafilter and carries out ultrafiltration, obtain the purification alkali protease containing compound stabilizer
Liquid finally adds composite antioxidant and mercapto-protective agent into the purification basic protein enzyme solution containing compound stabilizer, is contained
The stabilized enzyme formulations of alkali protease.
The alkali protease thick hair zymotic fluid is fermented by Pichia pastoris to be generated；
The weight percent of the composite flocculation agent and alkali protease thick hair zymotic fluid component are as follows: 75% protease thick hair
Zymotic fluid, 25% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 98% organic/inorganic polymer flocculants,
2% inorganic salts flocculant；
The organic/inorganic polymer flocculants are polyacrylamide；Inorganic salts are the mixing of aluminum sulfate, calcium sulfate, are mixed
Composition and division in a proportion example is 3:1；
The weight percent of the compound stabilizer is the 25% of basic protein enzyme solution, and compound stabilizer is trehalose and third
The mixture of glycol, mixed proportion 1:7；
The weight hundred of the purification basic protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 98% purification basic protein enzyme solution containing compound stabilizer, 3% composite antioxidant, surplus are that sulfydryl is protected
Protect agent；The weight hundred of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
Dividing the sum of ratio is 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is the mixture of cysteine, glutathione, mixed proportion 1:2.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stabilization enzyme containing alkali protease
Preparation is applied in cleaning product.
Alkali protease is shown in Table 1 using the method enzyme activity comparison before and after the processing of the present embodiment.
Embodiment 7
A kind of refining methd of the stable protease preparation of alkali protease, method particularly includes:
A kind of alkali protease thick hair zymotic fluid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then be sent into ultrafilter after addition compound stabilizer thereto and carry out ultrafiltration, obtain containing stable composition
The purification basic protein enzyme solution of agent, finally into the purification basic protein enzyme solution containing compound stabilizer add composite antioxidant and
Mercapto-protective agent obtains the stabilized enzyme formulations containing alkali protease.
The alkali protease thick hair zymotic fluid is fermented by bacillus lentus to be generated；
The weight percent of the composite flocculation agent and alkali protease thick hair zymotic fluid component are as follows: 55% protease thick hair
Zymotic fluid, 45% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 60% organic/inorganic polymer flocculants,
40% inorganic salts flocculant；
The organic/inorganic polymer flocculants are the mixing of polyacrylamide, aluminium polychloride, mixing ratio 1:5；Nothing
Machine salt is the mixing of aluminum sulfate and calcium sulfate, mixed proportion 1:1；
The weight percent of the compound stabilizer is the 4% of basic protein enzyme solution, and compound stabilizer is trehalose and third
The mixture of glycol, glycerine, mixed proportion 1:4:2；
The weight hundred of the purification basic protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 93% purification basic protein enzyme solution containing compound stabilizer, 2% composite antioxidant, surplus are that sulfydryl is protected
Protect agent；The weight hundred of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
Dividing the sum of ratio is 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is the mixture of cysteine, glutathione, mixed proportion 1:5.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stabilization enzyme containing alkali protease
Preparation is applied in cleaning product.
Alkali protease is shown in Table 1 using the method enzyme activity comparison before and after the processing of the present embodiment.
Embodiment 8
A kind of preparation process of the stabilized enzyme formulations containing alkali protease are as follows:
A kind of alkali protease thick hair zymotic fluid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then thereto after addition compound stabilizer, limpid alkali protease is obtained by filtering, plate compression
Liquid, then limpid basic protein enzyme solution is sent into ultrafilter and carries out ultrafiltration, obtain the purification alkali protease containing compound stabilizer
Liquid finally adds composite antioxidant and mercapto-protective agent into the purification basic protein enzyme solution containing compound stabilizer, is contained
The stabilized enzyme formulations of alkali protease.
The alkali protease thick hair zymotic fluid is fermented by Pichia pastoris to be generated；
The weight percent of the composite flocculation agent and alkali protease thick hair zymotic fluid component are as follows: 95% protease thick hair
Zymotic fluid, 5% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 5% organic/inorganic polymer flocculants,
95% inorganic salts flocculant；
The organic/inorganic polymer flocculants are the mixing of polyacrylamide, bodied ferric sulfate, mixing ratio 1:1；Nothing
Machine salt is the mixing of aluminum sulfate, calcium chloride, calcium sulfate, mixed proportion 1:1:1；
The weight percent of the compound stabilizer is the 2% of basic protein enzyme solution, and compound stabilizer is trehalose and third
The mixture of glycol, mixed proportion 1:3；
The weight hundred of the purification basic protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 97% purification basic protein enzyme solution containing compound stabilizer, 1.5% composite antioxidant, surplus are sulfydryl
Protective agent；The weight of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
The sum of percentage is 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is the mixture of cysteine, glutathione, mixed proportion 1:5.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stabilization enzyme containing alkali protease
Preparation is applied in cleaning product.
Alkali protease is shown in Table 1 using the method enzyme activity comparison before and after the processing of the present embodiment.
Embodiment 9
A kind of preparation process of the stabilized enzyme formulations containing alkali protease are as follows:
A kind of alkali protease thick hair zymotic fluid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then thereto after addition compound stabilizer, limpid alkali protease is obtained by filtering, plate compression
Liquid, then limpid basic protein enzyme solution is sent into ultrafilter and carries out ultrafiltration, obtain the purification alkali protease containing compound stabilizer
Liquid finally adds composite antioxidant and mercapto-protective agent into the purification basic protein enzyme solution containing compound stabilizer, is contained
The stabilized enzyme formulations of alkali protease.
The alkali protease thick hair zymotic fluid is generated by fermentation of bacillus subtilis；
The weight percent of the composite flocculation agent and alkali protease thick hair zymotic fluid component are as follows: 35% protease thick hair
Zymotic fluid, 65% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 50% organic/inorganic polymer flocculants,
50% inorganic salts flocculant；
The organic/inorganic polymer flocculants are the mixing of polyacrylamide, aluminium polychloride, mixing ratio 1:3；Nothing
Machine salt is the mixing of aluminum sulfate and calcium chloride, mixed proportion 3:1；
The weight percent of the compound stabilizer is the 3% of basic protein enzyme solution, and compound stabilizer is trehalose and third
The mixture of glycol, glycerine, mixed proportion 1:4:5；
The weight hundred of the purification basic protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 96% purification basic protein enzyme solution containing compound stabilizer, 2.5% composite antioxidant, surplus are sulfydryl
Protective agent；The weight of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
The sum of percentage is 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is glutathione.
Protease preparation is handled and prepared with technique by the above process, can be obtained the stabilization enzyme containing alkali protease
Preparation is applied in cleaning product.
Alkali protease is shown in Table 1 using the method enzyme activity comparison before and after the processing of this implementation.
Embodiment 10
A kind of preparation process of the stabilized enzyme formulations containing alkali protease are as follows:
A kind of alkali protease thick hair zymotic fluid obtains after filtering, suction filtration or plate compression after adding composite flocculation agent
Limpid basic protein enzyme solution, then thereto after addition compound stabilizer, limpid alkali protease is obtained by filtering, plate compression
Liquid, then limpid basic protein enzyme solution is sent into ultrafilter and carries out ultrafiltration, obtain the purification alkali protease containing compound stabilizer
Liquid finally adds composite antioxidant and mercapto-protective agent into the purification basic protein enzyme solution containing compound stabilizer, is contained
The stabilized enzyme formulations of alkali protease.
The alkali protease thick hair zymotic fluid is fermented by Bacillus licheniformis2709 to be generated；
The weight percent of the composite flocculation agent and alkali protease thick hair zymotic fluid component are as follows: 94% protease thick hair
Zymotic fluid, 6% composite flocculation agent；
The weight percent of contained component in the composite flocculation agent are as follows: 70% organic/inorganic polymer flocculants,
30% inorganic salts flocculant；
The organic/inorganic polymer flocculants are aluminium polychloride；Inorganic salts are the mixing of aluminum sulfate and calcium sulfate, are mixed
Composition and division in a proportion example is 4:1；
The weight percent of the compound stabilizer is the 7% of basic protein enzyme solution, and compound stabilizer is trehalose and third
The mixture of triol, mixed proportion 1:1；
The weight hundred of the purification basic protein enzyme solution containing compound stabilizer, composite antioxidant and mercapto-protective agent
Point ratio are as follows: the 94% purification basic protein enzyme solution containing compound stabilizer, 0.5% composite antioxidant, surplus are sulfydryl
Protective agent；The weight of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
The sum of percentage is 100%.
The composite antioxidant is the mixing of vitamin C and sodium thiosulfate, mixed proportion 1:1；
The mercapto-protective agent is the mixture of cysteine, glutathione, mixed proportion 3:1.
Protease preparation, the stability egg containing alkali protease of acquisition are handled and prepared with technique by the above process
White enzyme preparation is in cleaning product.
Alkali protease is shown in Table 1 using the method enzyme activity comparison before and after the processing of this implementation.
Table 1 is the purification that the stable protease preparation containing 2709 alkali proteases of the invention is used in embodiment 1-10
The enzyme of method preparation compares (folin's methods measurement) with the enzyme activity of this method is not used.
Table 1:
It is compared using enzyme prepared by the present invention with the enzyme activity of this method is not used
It is composite antioxidant, mercapto-protective agent, organic/inorganic polymer flocculants in above-mentioned 1-10 embodiment, inorganic
Salt, non-reducible disaccharide and polyalcohol are not limited to selected substance, and composite antioxidant can arbitrarily replace with vitamin A, dimension
The mixing of one of raw element E, vitamin C, ascorbic acid, sodium thiosulfate or several arbitrary proportions；Mercapto-protective agent is
One of cysteine, glutathione, 2 mercapto ethanol, dithiothreitol (DTT), dithioerythritol or several arbitrary proportions
Mixing；Organic/inorganic polymer flocculants are polyacrylamide PAM, PB1 gene, bodied ferric sulfate SPFS or poly-
Close the mixing of one of aluminium chloride ferrum PAFC or several arbitrary proportions；Inorganic salts are calcium sulfate, calcium chloride, aluminum sulfate, sulphur
The mixing of one of sour sodium, sodium chloride, magnesium sulfate or several arbitrary proportions；Non-reducible disaccharide is sucrose, in trehalose
The mixing of one or several kinds of arbitrary proportions；Polyalcohol is one of propylene glycol, dipropylene glycol, glycerine or several any
The mixing of ratio.
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art,
It is still possible to modify the technical solutions described in the foregoing embodiments, or part of technical characteristic is carried out etc.
With replacement, all within the spirits and principles of the present invention, any modification, equivalent replacement, improvement and so on should be included in this
Within the protection scope of invention.
Claims (10)
1. the stabilized enzyme formulations containing alkalinity or neutral proteinase, it is characterised in that: the stabilized enzyme formulations are by the following method
It is made；Specially alkalinity or natural protease thick fermentation liquid are after composite flocculation agent flocculation treatment, by filtering, suction filtration or plate
Frame filters pressing obtains limpid alkalinity or neutral protein enzyme solution, then compound stabilizer is added thereto and carries out ultrafiltration or crosses ion exchange
Column purification；Alkalinity or neutral protein enzyme solution and composite antioxidant and mercapto-protective agent after purification are configured to stabilized enzyme formulations.
2. the stabilized enzyme formulations according to claim 1 containing alkalinity or neutral proteinase, it is characterised in that: the compound wadding
Solidifying agent is the mixture of organic/inorganic polymer and inorganic salts, the weight percent of component contained by the composite flocculation agent are as follows:
5%~98% organic/inorganic polymer flocculants, surplus are inorganic salts flocculant；The organic/inorganic polymer is propylene
Any one in amide PAM, PB1 gene, bodied ferric sulfate SPFS or polyaluminum ferric chloride PAFC, inorganic salts are
The mixing of one of calcium sulfate, calcium chloride, aluminum sulfate, sodium sulphate, sodium chloride, magnesium sulfate or several arbitrary proportions；It is described
The compound stabilizer of addition is the 0.1%~40% of limpid alkalinity or neutral proteinase fermentation liquid weight；The compound stabilizer
For the mixture that non-reducible disaccharide and polyalcohol form, the weight ratio of non-reducible disaccharide and polyalcohol is 1:1~10, wherein non-go back
Former disaccharides is one of sucrose, trehalose or the mixing of their arbitrary proportions, and polyalcohol is propylene glycol, dipropylene glycol, third
The mixing of one of triol or several arbitrary proportions；Purification alkalinity or neutral protein enzyme solution, composite antioxidant and
The weight percent of mercapto-protective agent are as follows: 35%~98% purification basic protein enzyme solution, 0.01%~5% compound anti-oxidation
Agent, surplus are mercapto-protective agent；The composite antioxidant is VitAVitE, in ascorbic acid, sodium thiosulfate
The mixing of one or several kinds of arbitrary proportions；The mercapto-protective agent is cysteine, glutathione, 2 mercapto ethanol, two sulphur
The mixing of one or more of threitol, dithioerythritol arbitrary proportion.
3. the preparation process of the stabilized enzyme formulations as claimed in claim 1 or 2 containing alkalinity or neutral proteinase, it is characterised in that:
Alkalinity or natural protease thick fermentation liquid are carried out using the composite flocculation agent that organic/inorganic polymer and inorganic salts are formulated
Flocculation treatment obtains limpid alkalinity or neutral protein enzyme solution by filtering, suction filtration or plate compression, then is added thereto multiple
Stabilizer is closed, is refining to obtain the alkalinity of the purification containing compound stabilizer or neutral protein enzyme solution through ultrafiltration or excessively ion exchange column；So
After will containing compound stabilizer purification alkalinity or neutral protein enzyme solution be hybridly prepared into composite antioxidant and mercapto-protective agent
Stabilized enzyme formulations containing alkalinity or neutral proteinase.
4. the preparation process of the stabilized enzyme formulations according to claim 3 containing alkalinity or neutral proteinase, it is characterised in that: institute
It states alkalinity or natural protease thick fermentation liquid is by Bacillus bacteria or yeast fermenting and producing, wherein Bacillus bacteria refers to starch
Bacillus, bacillus lentus, actinomyces, aspergillus terricola, bacillus subtilis or bacillus licheniformis.
5. the preparation process of the stabilized enzyme formulations according to claim 3 containing alkalinity or neutral proteinase, it is characterised in that: institute
State composite flocculation agent and alkalinity or natural protease thick fermentation liquid component weight percent are as follows: 35%~99.5% alkalinity or
Natural protease thick fermentation liquid, 0.5%~65% composite flocculation agent.
6. the preparation process of the stabilized enzyme formulations according to claim 3 containing alkalinity or neutral proteinase, it is characterised in that: institute
Stating composite flocculation agent is formulated by organic/inorganic polymer and inorganic salts, the weight of component contained by the composite flocculation agent
Percentage are as follows: 5%~98% organic/inorganic polymer flocculants, 2%~95% inorganic salts flocculant；Organic/the nothing
Machine polymer is one in polyacrylamide PAM, PB1 gene, bodied ferric sulfate SPFS or polyaluminum ferric chloride PAFC
The mixing of kind or several arbitrary proportions；The inorganic salts are calcium sulfate, calcium chloride, aluminum sulfate, sodium sulphate, sodium chloride, sulfuric acid
The mixing of one of magnesium or several arbitrary proportions.
7. the preparation process of the stabilized enzyme formulations according to claim 3 containing alkalinity or neutral proteinase, it is characterised in that: institute
The compound stabilizer for stating addition is the 0.1%~40% of limpid alkalinity or neutral protein enzyme solution weight, and the compound stabilizer is
The mixture of non-reducible disaccharide and polyalcohol composition, wherein the weight ratio of non-reducible disaccharide and polyalcohol is 1:1~10, described non-
Reduction disaccharides is the mixing of one of sucrose, trehalose or several arbitrary proportions；The polyalcohol is propylene glycol, dipropyl two
The mixing of one of alcohol, glycerine or several arbitrary proportions.
8. the preparation process of the stabilized enzyme formulations according to claim 3 containing alkalinity or neutral proteinase, it is characterised in that: institute
State the weight percent of purification alkalinity or neutral protein enzyme solution, composite antioxidant and mercapto-protective agent containing compound stabilizer
Are as follows: 35%~98% compound anti-oxidation that alkaline or neutral protein enzyme solution, 0.01%~5% are refined containing compound stabilizer
Agent, surplus are mercapto-protective agent, purification alkalinity or neutral protein enzyme solution, composite antioxidant and sulfydryl containing compound stabilizer
The sum of protectant weight percent is 100%.
9. the preparation process of the stabilized enzyme formulations according to claim 3 containing alkalinity or neutral proteinase, it is characterised in that:
The composite antioxidant is one of VitAVitE, vitamin C, ascorbic acid, sodium thiosulfate or several
The mixing of arbitrary proportion；The mercapto-protective agent is cysteine, glutathione, 2 mercapto ethanol, dithiothreitol (DTT), two sulphur
The mixing of one of antierythrite or several arbitrary proportions.
10. the application of the stabilized enzyme formulations as claimed in claim 1 or 2 containing alkalinity or neutral proteinase, it is characterised in that: institute
The stabilized enzyme formulations containing alkalinity or neutral proteinase are stated to be applied in detergent product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910481004.9A CN110229803A (en) | 2019-06-04 | 2019-06-04 | Stabilized enzyme formulations and its preparation process and application containing alkalinity or neutral proteinase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910481004.9A CN110229803A (en) | 2019-06-04 | 2019-06-04 | Stabilized enzyme formulations and its preparation process and application containing alkalinity or neutral proteinase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110229803A true CN110229803A (en) | 2019-09-13 |
Family
ID=67859229
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910481004.9A Pending CN110229803A (en) | 2019-06-04 | 2019-06-04 | Stabilized enzyme formulations and its preparation process and application containing alkalinity or neutral proteinase |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110229803A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111117790A (en) * | 2020-02-25 | 2020-05-08 | 沈阳垚采科技有限公司 | Preparation method of washing-free cleaning agent |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102676482A (en) * | 2012-05-11 | 2012-09-19 | 河南省南街村（集团）有限公司 | Method for producing alkali protease from highland bacillus |
| CN108728258A (en) * | 2018-07-19 | 2018-11-02 | 杜永卫 | A kind of portable biometric enzyme stain removal agent and preparation method thereof |
-
2019
- 2019-06-04 CN CN201910481004.9A patent/CN110229803A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102676482A (en) * | 2012-05-11 | 2012-09-19 | 河南省南街村（集团）有限公司 | Method for producing alkali protease from highland bacillus |
| CN108728258A (en) * | 2018-07-19 | 2018-11-02 | 杜永卫 | A kind of portable biometric enzyme stain removal agent and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 刘秉智等: "液体洗涤剂中酶稳定性的研究", 《渭南师范学院学报》 * |
| 牟占军: "2709碱性蛋白酶菌体絮凝分离的研究", 《内蒙古工业大学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111117790A (en) * | 2020-02-25 | 2020-05-08 | 沈阳垚采科技有限公司 | Preparation method of washing-free cleaning agent |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102732589B (en) | Method for treating threonine mother liquor | |
| CN102719510B (en) | Amino acid fermentation bacteria utilization method | |
| CN110205314A (en) | Stabilized enzyme formulations and its preparation process and application containing 2709 alkali proteases | |
| CN103937846B (en) | A kind of method utilizing cotton cake dregs to prepare Moriamin S | |
| CN109504719A (en) | A method of improving glutamic acid acid production rate and recovery rate | |
| CN102533884B (en) | Method for cleanly producing glutamic acid, gamma-polyglutamic acid and organic fertilizer | |
| CN102701507B (en) | Method for treating high-concentration wastewater of glutamic acid | |
| CN104726527A (en) | Preparation technology for producing collagen through enzymolysis of fish scales | |
| CN110547458A (en) | Preparation method of enzymolysis pearl powder | |
| CN103087126A (en) | Preparation method of zhongshengmycin bulk drug | |
| CN102887946A (en) | Method for extracting sericin from cocoon cooking wastewater by physical method | |
| CN110229803A (en) | Stabilized enzyme formulations and its preparation process and application containing alkalinity or neutral proteinase | |
| CN103772532A (en) | Method for processing skim rubber | |
| CN108285911A (en) | A kind of technique of fermentation extraction l-Isoleucine | |
| CN111073941A (en) | Preparation process of sandalwood polypeptide | |
| CN114015739A (en) | Method for preparing liquid collagen peptide from tilapia skin | |
| CN114250008A (en) | Biological cleaning agent capable of degrading printing ink coating and preparation method thereof | |
| CN103642885A (en) | Method for producing wheat biological active peptide by fermentation | |
| CN103864954A (en) | Method for extracting peanut meal polysaccharide | |
| CN102766671B (en) | Method for extracting proteins from vitamin C fermented mash | |
| CN104450840A (en) | Production process of bovine bone peptone | |
| CN104045686A (en) | Method for treating bacillus subtilis antimicrobial lipopeptide fermentation broth | |
| Guo-qing et al. | Partitioning and purification of extracellular β-1, 3-1, 4-glucanase in aqueous two-phase systems | |
| CN109536556A (en) | A kind of preparation method of homogeneity green alga active peptide | |
| CN111424067B (en) | Method for extracting low-component sea cucumber bioactive peptide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190913 |
|
| RJ01 | Rejection of invention patent application after publication |