CN112125747A - Microbial agent for enzymolysis of soybean phospholipid - Google Patents
Microbial agent for enzymolysis of soybean phospholipid Download PDFInfo
- Publication number
- CN112125747A CN112125747A CN202010497333.5A CN202010497333A CN112125747A CN 112125747 A CN112125747 A CN 112125747A CN 202010497333 A CN202010497333 A CN 202010497333A CN 112125747 A CN112125747 A CN 112125747A
- Authority
- CN
- China
- Prior art keywords
- enzymolysis
- soybean phospholipid
- microbial agent
- soybean
- phospholipid
- 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
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 title claims abstract description 88
- 239000008347 soybean phospholipid Substances 0.000 title claims abstract description 75
- 230000000813 microbial effect Effects 0.000 title claims abstract description 46
- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 44
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 25
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 25
- 230000012010 growth Effects 0.000 claims abstract description 24
- 235000013379 molasses Nutrition 0.000 claims abstract description 24
- 239000001963 growth medium Substances 0.000 claims abstract description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 229940083466 soybean lecithin Drugs 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 14
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 10
- LFVGISIMTYGQHF-UHFFFAOYSA-N ammonium dihydrogen phosphate Chemical compound [NH4+].OP(O)([O-])=O LFVGISIMTYGQHF-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229910000387 ammonium dihydrogen phosphate Inorganic materials 0.000 claims abstract description 10
- 239000004202 carbamide Substances 0.000 claims abstract description 10
- 235000019837 monoammonium phosphate Nutrition 0.000 claims abstract description 10
- 239000006012 monoammonium phosphate Substances 0.000 claims abstract description 10
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 10
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 10
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 10
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims abstract description 10
- 230000001737 promoting effect Effects 0.000 claims abstract description 10
- 239000011780 sodium chloride Substances 0.000 claims abstract description 10
- 239000011573 trace mineral Substances 0.000 claims abstract description 7
- 235000013619 trace mineral Nutrition 0.000 claims abstract description 7
- 229940088594 vitamin Drugs 0.000 claims abstract description 7
- 229930003231 vitamin Natural products 0.000 claims abstract description 7
- 235000013343 vitamin Nutrition 0.000 claims abstract description 7
- 239000011782 vitamin Substances 0.000 claims abstract description 7
- 150000003904 phospholipids Chemical class 0.000 claims description 23
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims description 18
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 18
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 16
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 16
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 13
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 10
- JVXHQHGWBAHSSF-UHFFFAOYSA-L 2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate;hydron;iron(2+) Chemical compound [H+].[H+].[Fe+2].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O JVXHQHGWBAHSSF-UHFFFAOYSA-L 0.000 claims description 9
- 102000003670 Carboxypeptidase B Human genes 0.000 claims description 9
- 108090000087 Carboxypeptidase B Proteins 0.000 claims description 9
- 102000016938 Catalase Human genes 0.000 claims description 9
- 108010053835 Catalase Proteins 0.000 claims description 9
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims description 9
- 102000057297 Pepsin A Human genes 0.000 claims description 9
- 108090000284 Pepsin A Proteins 0.000 claims description 9
- 229930003427 Vitamin E Natural products 0.000 claims description 9
- 229910052802 copper Inorganic materials 0.000 claims description 9
- 239000010949 copper Substances 0.000 claims description 9
- 235000019152 folic acid Nutrition 0.000 claims description 9
- 239000011724 folic acid Substances 0.000 claims description 9
- 229960000304 folic acid Drugs 0.000 claims description 9
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims description 9
- 229940111202 pepsin Drugs 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 235000019165 vitamin E Nutrition 0.000 claims description 9
- 229940046009 vitamin E Drugs 0.000 claims description 9
- 239000011709 vitamin E Substances 0.000 claims description 9
- 229940045997 vitamin a Drugs 0.000 claims description 9
- YTSDVWYUJXKXCC-UHFFFAOYSA-L zinc;2-[2-[carboxylatomethyl(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetate Chemical compound [Zn+2].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O YTSDVWYUJXKXCC-UHFFFAOYSA-L 0.000 claims description 9
- 239000002994 raw material Substances 0.000 claims description 8
- 235000012424 soybean oil Nutrition 0.000 claims description 8
- 239000003549 soybean oil Substances 0.000 claims description 8
- 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 claims description 7
- 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 claims description 7
- 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 claims description 7
- 238000012258 culturing Methods 0.000 claims description 7
- 235000019155 vitamin A Nutrition 0.000 claims description 7
- 239000011719 vitamin A Substances 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 6
- 235000015112 vegetable and seed oil Nutrition 0.000 claims description 6
- 239000008158 vegetable oil Substances 0.000 claims description 6
- 229930002875 chlorophyll Natural products 0.000 claims description 5
- 235000019804 chlorophyll Nutrition 0.000 claims description 5
- 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 claims description 5
- 241000894006 Bacteria Species 0.000 claims description 4
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 claims description 4
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 claims description 4
- 229930003471 Vitamin B2 Natural products 0.000 claims description 4
- 229960002477 riboflavin Drugs 0.000 claims description 4
- 238000010025 steaming Methods 0.000 claims description 4
- 235000019164 vitamin B2 Nutrition 0.000 claims description 4
- 239000011716 vitamin B2 Substances 0.000 claims description 4
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 4
- 235000019484 Rapeseed oil Nutrition 0.000 claims description 3
- 235000005687 corn oil Nutrition 0.000 claims description 3
- 239000002285 corn oil Substances 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 235000019483 Peanut oil Nutrition 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 239000000312 peanut oil Substances 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 12
- 235000015097 nutrients Nutrition 0.000 abstract description 11
- 230000000694 effects Effects 0.000 abstract description 10
- 235000002639 sodium chloride Nutrition 0.000 abstract description 10
- 239000002689 soil Substances 0.000 abstract description 10
- 102000004190 Enzymes Human genes 0.000 abstract description 8
- 108090000790 Enzymes Proteins 0.000 abstract description 8
- 244000005700 microbiome Species 0.000 abstract description 8
- 230000008635 plant growth Effects 0.000 abstract description 5
- 230000009286 beneficial effect Effects 0.000 abstract description 4
- 235000013399 edible fruits Nutrition 0.000 abstract description 4
- 230000004060 metabolic process Effects 0.000 abstract description 4
- 235000013877 carbamide Nutrition 0.000 abstract 1
- 244000241257 Cucumis melo Species 0.000 description 13
- 239000000126 substance Substances 0.000 description 12
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 11
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 10
- 239000003337 fertilizer Substances 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 7
- 229940088598 enzyme Drugs 0.000 description 7
- 235000016709 nutrition Nutrition 0.000 description 6
- 230000035764 nutrition Effects 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 238000011282 treatment Methods 0.000 description 6
- 244000046052 Phaseolus vulgaris Species 0.000 description 5
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 5
- 229930003270 Vitamin B Natural products 0.000 description 5
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 235000019156 vitamin B Nutrition 0.000 description 5
- 239000011720 vitamin B Substances 0.000 description 5
- 235000010469 Glycine max Nutrition 0.000 description 4
- 244000052616 bacterial pathogen Species 0.000 description 4
- 229920002521 macromolecule Polymers 0.000 description 4
- 238000003828 vacuum filtration Methods 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 244000068988 Glycine max Species 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 3
- 239000010779 crude oil Substances 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 235000019198 oils Nutrition 0.000 description 3
- 239000011574 phosphorus Substances 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 2
- 235000009847 Cucumis melo var cantalupensis Nutrition 0.000 description 2
- 108010026389 Gramicidin Proteins 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 108010040201 Polymyxins Proteins 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- RKLXDNHNLPUQRB-TVJUEJKUSA-N chembl564271 Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]2C(C)SC[C@H](N[C@@H](CC(N)=O)C(=O)NC(=O)[C@@H](NC2=O)CSC1C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NC(=C)C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@H]1NC(=O)C(=C\C)/NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H]2NC(=O)CNC(=O)[C@@H]3CCCN3C(=O)[C@@H](NC(=O)[C@H]3N[C@@H](CC(C)C)C(=O)NC(=O)C(=C)NC(=O)CC[C@H](NC(=O)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC=4C5=CC=CC=C5NC=4)CSC3)C(O)=O)C(C)SC2)C(C)C)C(C)SC1)C1=CC=CC=C1 RKLXDNHNLPUQRB-TVJUEJKUSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000008157 edible vegetable oil Substances 0.000 description 2
- 230000001804 emulsifying effect Effects 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 229960004905 gramicidin Drugs 0.000 description 2
- ZWCXYZRRTRDGQE-SORVKSEFSA-N gramicidina Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CC=3C4=CC=CC=C4NC=3)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CC=3C4=CC=CC=C4NC=3)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CC=3C4=CC=CC=C4NC=3)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](C(C)C)NC(=O)[C@H](C)NC(=O)[C@H](NC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](NC=O)C(C)C)CC(C)C)C(=O)NCCO)=CNC2=C1 ZWCXYZRRTRDGQE-SORVKSEFSA-N 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000000787 lecithin Substances 0.000 description 2
- 229940067606 lecithin Drugs 0.000 description 2
- 235000010445 lecithin Nutrition 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 229960000988 nystatin Drugs 0.000 description 2
- VQOXZBDYSJBXMA-NQTDYLQESA-N nystatin A1 Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/CC/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 VQOXZBDYSJBXMA-NQTDYLQESA-N 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 230000002028 premature Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 108010082567 subtilin Proteins 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 description 1
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 1
- 241000208838 Asteraceae Species 0.000 description 1
- 229920001268 Cholestyramine Polymers 0.000 description 1
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000009144 enzymatic modification Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- -1 inositol phospholipids Chemical class 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000037257 muscle growth Effects 0.000 description 1
- 230000008271 nervous system development Effects 0.000 description 1
- 239000012454 non-polar solvent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 239000004455 soybean meal Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Pest Control & Pesticides (AREA)
- Soil Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Fertilizers (AREA)
Abstract
The invention discloses a microbial agent for enzymolysis of soybean phospholipid, which is prepared by the following steps: adding various enzymes into the soybean phospholipid liquid for enzymolysis to obtain enzymolysis soybean phospholipid; preparing enzymolysis soybean phospholipid, molasses, monoammonium phosphate, urea, monopotassium phosphate, vitamins, trace elements, sodium chloride and water into a culture medium according to a certain amount; inoculating the bacillus subtilis seed solution into a culture medium, and fermenting to obtain the microbial agent for enzymolysis of soybean lecithin. The microbial agent prepared by the invention can promote the propagation of farmland soil microorganisms, increase the number of beneficial floras in soil, improve the soil environment and prevent soil hardening; can provide organic nutrients which can be directly absorbed and utilized for crops, improve the taste of fruits and increase appearance market phase; can directly regulate the metabolism of plants and stimulate the growth of crops; can also promote the growth of plant roots, and has the effects of promoting flowers and fruits and increasing the yield and quality of crops.
Description
Technical Field
The invention relates to the technical field of biological fertilizers, in particular to a microbial agent for enzymolysis of soybean lecithin.
Background
In the process of soybean production and separation, the first soybean meal is separated, and the rest is grease. Crude oil, which is our crude oil, is extracted from the beginning, and the soybean phospholipids are in the crude oil. During the oil hydration process, the soybean phospholipid is partially separated, and the rest is the edible oil. So the soybean phospholipids are the products extracted from the oil foot/soapstock in the production process of soybean refined edible oil. Soybean lecithin is an ester composed of glycerol, fatty acid, choline or cholestyramine, and is soluble in oil and nonpolar solvent. Soy phospholipids are complex in composition and contain mostly lecithin (about 34.2%), cephalins (about 19.7%), inositol phospholipids (about 16.0%), phosphatidylserine (about 15.8%), phosphatidic acid (about 3.6%) and other phospholipids (about 10.7%). The soybean lecithin not only has stronger emulsification, moistening and dispersing functions, but also plays an important role in promoting fat metabolism in vivo, muscle growth, nervous system development, oxidation damage resistance in vivo and the like. Therefore, soybean phospholipids in soybean are generally used for physiological health care at present.
The soybean lecithin is in a faint yellow to brown viscous fluid or solid powder after enzymolysis, has better stability because unsaturated fatty acid bonds on beta positions are removed by enzyme modification, is not easy to oxidize, and has a prolonged storage life, and because of strong emulsifying property, the soybean lecithin acts on the same product, the addition amount of the soybean lecithin is about 1/10 of common lecithin, and the emulsifying property is hardly influenced under the conditions of salt and calcium salt. Therefore, enzymatically hydrolyzed soybean phospholipids are generally used as an emulsifier. At present, no report about the utilization of enzymatic soybean phospholipid as a fertilizer is found.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a microbial agent for enzymolysis of soybean lecithin. Can promote the increase of chlorophyll content in crops and promote the growth of stems and tendrils of the crops.
The invention is realized by the following technical scheme:
the invention provides a microbial agent for enzymolysis of soybean phospholipid, which is prepared by the following steps:
(1) adding catalase, pepsin and carboxypeptidase B into the soybean phospholipid liquid, adjusting the pH value to 7.0, performing enzymolysis for 48 hours, heating to 80 ℃, and stopping enzymolysis reaction to obtain enzymolysis soybean phospholipid;
(2) preparing a culture medium according to the following raw materials in percentage by weight: 20-30% of enzymolysis soybean phospholipid, 10-20% of molasses, 1% of monoammonium phosphate, 1.6% of urea, 1% of monopotassium phosphate, 0.6% of vitamin, 0.401% of trace elements, 1-2% of sodium chloride and the balance of water;
(3) inoculating the bacillus subtilis seed solution into a culture medium according to the inoculation amount of 1-2%, adjusting the pH to 6.0-6.5, and culturing for 32-48 hours to obtain the microbial agent for enzymolysis of soybean lecithin.
Preferably, in the step (1), the soybean phospholipid solution is prepared by the following method:
1) dehydrating the soybean oil residue to obtain crude phospholipid;
2) adding acetone with equal mass into the crude phospholipid, stirring, performing solid-liquid separation after stirring is finished, and recovering the acetone to obtain crude phospholipid powder;
3) adding 30% hydrogen peroxide of 1% of the weight of the material into the crude phospholipid powder, fully stirring and decoloring, and steaming out the hydrogen peroxide to obtain refined soybean phospholipid;
4) dissolving refined soybean phospholipid in vegetable oil to obtain soybean phospholipid solution.
Preferably, the adding amount of the refined soybean phospholipid and the vegetable oil is 10-5: 1; the vegetable oil is one or more of soybean oil, corn oil, rapeseed oil and peanut oil.
Preferably, in the step (1), the addition amount of the catalase is 0.6% of the weight of the soybean lecithin liquid, the addition amount of the pepsin is 0.3% of the weight of the soybean lecithin liquid, the addition amount of the carboxypeptidase B is 0.1% of the weight of the soybean lecithin liquid, and the enzymolysis temperature is 25-35 ℃. The invention adopts a plurality of enzymes for enzymolysis, and can enhance a plurality of hydrolysis and redox reactions. Has the synergistic metabolism of endogenous enzyme and exogenous enzyme, and can accelerate enzymolysis.
Preferably, in the step (2), the molasses is cane molasses, the brix of the molasses is 80-90 Bx, and the sugar content is more than 50%.
Preferably, in the step (2), the vitamins include vitamin a, vitamin B2, vitamin E and folic acid, and the mass ratio of the vitamin a to the vitamin B2 to the vitamin E to the folic acid is 1.7: 1: 2: 1.3.
preferably, in the step (2), the trace elements include EDTA-iron, EDTA-manganese, EDTA-copper and EDTA-zinc, and the mass ratio of EDTA-iron, EDTA-manganese, EDTA-copper and EDTA-zinc is 2: 1: 0.01: 1.
preferably, in the step (3), the number of effective viable bacteria in the microbial agent for enzymolysis of soybean phospholipid is more than or equal to 20.0 multiplied by 109cfu/mL
The bacillus subtilis is purchased from Shandong agriculture and fertilizer industry science and technology Limited. Active substances such as subtilin, polymyxin, nystatin, gramicidin and the like generated in the growth process of bacillus subtilis have obvious inhibiting effect on pathogenic bacteria or conditional pathogenic bacteria of endogenous infection.
The invention provides application of the microbial agent for enzymolysis of soybean phospholipid in promoting growth of crops.
Preferably, the promoting of the growth of the crops is promoting of the increase of the chlorophyll content in the crops and promoting of the growth of the stems and tendrils of the crops.
The invention has the beneficial effects that:
1. according to the invention, the bean phospholipid is subjected to enzymolysis by compounding a plurality of enzymes, and the enzymolysis bean phospholipid is used in the formula of the microbial agent to prepare the microbial agent, so that the propagation of microorganisms in farmland soil can be promoted, the number of beneficial floras in the soil can be increased, the soil environment can be improved, and the soil hardening phenomenon can be prevented; can provide organic nutrients which can be directly absorbed and utilized for crops, improve the taste of fruits and increase appearance market phase; can directly regulate the metabolism of plants and stimulate the growth of crops; can also promote the growth of plant roots, and has the effects of promoting flowers and fruits and increasing the yield and quality of crops.
2. The invention takes a plurality of substances such as zymolytic bean phospholipid, molasses and the like as culture media, inoculates bacillus subtilis for fermentation, the zymolytic bean phospholipid and molasses can provide nutrition for the bacillus subtilis, and the bacillus subtilis can decompose macromolecular substances in the zymolytic bean phospholipid and molasses to provide a large amount of organic phosphorus and small molecular substances which are easy to absorb by plants, so that the fertilizer nutrition is richer and easier to absorb, the growth of plant root systems is promoted, the growth of main roots is increased, lateral roots and capillary roots are increased, plants are kept healthy and strong in the whole growth period, and the stress resistance and the premature senility resistance of the plants are enhanced.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
As described in the background art, the microbial fertilizer refers to a product containing a living body of a specific microorganism, which is applied to agricultural production, and increases the supply of plant nutrients or promotes the growth of plants, increases the yield, improves the quality of agricultural products and the agricultural ecological environment through the life activities of the microorganism contained therein. The life activity of microorganisms in soil must have enough nutrient substances as a guarantee, and the lack of nutrition can cause the rapid death and decay of functional microorganisms, so that the field application effect of the microbial fertilizer is poor. The bacillus subtilis generally takes beef extract, peptone and the like as culture media, and is mixed with nutrient substances after fermentation to serve as a microbial agent, while crops slowly absorb the nutrient substances, so that fertilizer waste is caused, the nutrient substances cannot be kept up as nutrients, and the growth is slow.
Based on the microbial agent, the microbial agent for enzymolysis of soybean phospholipid provided by the invention can continuously provide sufficient nutrition for microorganisms and keep the activity of the microorganisms. Preparing a culture medium according to the following raw materials in percentage by weight: 20-30% of enzymolysis soybean phospholipid, 10-20% of molasses, 1% of monoammonium phosphate, 1.6% of urea, 1% of monopotassium phosphate, 0.6% of vitamin, 0.401% of trace elements, 1-2% of sodium chloride and the balance of water. The culture medium can culture the bacillus subtilis and provide nutrients required by growth of crops, the bacillus subtilis decomposes macromolecular substances in enzymolysis soybean phospholipid and molasses in a seed expanding process to provide a large amount of organic phosphorus and small molecular substances which are easy to absorb by plants, so that the fertilizer nutrition is richer and easier to absorb, the growth of plant root systems is promoted, the growth of main roots is increased, lateral roots and capillary roots are increased, the plants are kept healthy and strong in the whole growth period, and the stress resistance and the premature senility resistance of the plants are enhanced.
When the microbial agent for enzymolysis of the soybean phospholipid is applied, active substances such as subtilin, polymyxin, nystatin, gramicidin and the like generated in the growth process of bacillus subtilis have obvious inhibiting effect on pathogenic bacteria or conditional pathogenic bacteria of endogenous infection; the molasses contains rich sugar, organic matters and mineral substances, and provides sufficient nutrient substances for the propagation of the bacillus subtilis and the soil beneficial bacteria. Meanwhile, the bacillus subtilis decomposes macromolecular substances in the enzymolysis soybean phospholipid and molasses in the seed expanding process, so that the fertilizer is richer in nutrition and easier to absorb as a large amount of organic phosphorus and small molecular substances which are easy to absorb by plants are provided, multiple purposes are achieved, nutrients are provided for crops in an all-round manner, and the growth of the crops is facilitated.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments. If the experimental conditions not specified in the examples are specified, the conditions are generally conventional or recommended by the reagent company; reagents, consumables, and the like used in the following examples are commercially available unless otherwise specified.
Description of the drawings: catalase was purchased from Zhengzhou Qiwashington chemical products Co., Ltd, and the enzyme activity was 5000-10000U/mg;
the pepsin is purchased from Kafen Biotech limited, Guangzhou, and the enzyme activity is 10000U/mg;
carboxypeptidase B is purchased from Shenzhenzhen Asteraceae Biotech Limited, and the enzyme activity is 170U/mg;
bacillus subtilis is purchased from Shandong agriculture and industry science and technology Limited.
Example 1: preparation of soybean phospholipid liquid
1) 500g of soybean oil residue is taken as water, most of the water is removed by vacuum filtration at normal temperature, and then a rotary evaporator is used for further concentrating and evaporating water at the temperature of 80 ℃ to obtain about 300g of crude phospholipid.
2) Adding 300g of crude phospholipid into a 2000ml conical flask, adding 300g of acetone, stirring for 4h at the temperature of 25 ℃, standing for layering, separating solid from liquid, performing vacuum filtration, recovering acetone solution, and performing vacuum drying on the obtained crude phospholipid powder.
3) Putting 200g of crude phospholipid powder into a 2000ml conical flask, adding 1g of hydrogen peroxide with the mass fraction of 30%, stirring for 2 hours at 80 ℃, and steaming out the hydrogen peroxide to obtain the refined soybean phospholipid.
4) 150g of refined soybean phospholipids were dissolved in 10g of soybean oil and 5g of corn oil to obtain a soybean phospholipid solution.
Example 2: preparation of soybean phospholipid liquid
1) 500g of soybean oil residue is taken as water, most of the water is removed by vacuum filtration at normal temperature, and then a rotary evaporator is used for further concentrating and evaporating water at the temperature of 70 ℃ to obtain about 300g of crude phospholipid.
2) Adding 300g of crude phospholipid into a 2000ml conical flask, adding 300g of acetone, stirring for 4h at the temperature of 25 ℃, standing for layering, separating solid from liquid, performing vacuum filtration, recovering acetone solution, and performing vacuum drying on the obtained crude phospholipid powder.
3) Putting 100g of crude phospholipid powder into a 2000ml conical flask, adding 1g of hydrogen peroxide with the mass fraction of 30%, stirring for 2 hours at 80 ℃, and steaming out the hydrogen peroxide to obtain the refined soybean phospholipid.
4) 50g of refined soybean phospholipid was dissolved in 4g of rapeseed oil and 6g of soybean oil to obtain a soybean phospholipid solution.
Example 3
(1) To 30g of the soybean phospholipid solution prepared in example 1, 0.18g of catalase, 0.09g of pepsin, and 0.03g of carboxypeptidase B were added, the pH was adjusted to 7.0, the enzymatic hydrolysis temperature was 25 ℃, and after 48 hours of enzymatic hydrolysis, the enzymatic hydrolysis reaction was stopped by heating to 80 ℃ to obtain enzymatically hydrolyzed soybean phospholipid.
(2) Preparing a culture medium according to the following raw materials in percentage by weight: 20g of enzymolysis soybean phospholipid, 10g of molasses, 1g of monoammonium phosphate, 1.6g of urea, 1g of monopotassium phosphate, 0.17g of vitamin A, 20.1g of vitamin B, 0.2g of vitamin E, 0.13g of folic acid, 0.2g of EDTA-iron, 0.1g of EDTA-manganese, 0.001g of EDTA-copper and 0.1g of EDTA-zinc, 1g of sodium chloride and 64.5g of water.
(3) Inoculating 1g of bacillus subtilis seed solution into a culture medium, adjusting the pH value to be 6.0-6.5, and culturing for 48 hours to obtain 100g of microbial agent for enzymolysis of soybean phospholipid.
Example 4
(1) To 30g of the soybean phospholipid solution prepared in example 1, 0.18g of catalase, 0.09g of pepsin, and 0.03g of carboxypeptidase B were added, the pH was adjusted to 7.0, the enzymatic hydrolysis temperature was 30 ℃, and after 48 hours of enzymatic hydrolysis, the enzymatic hydrolysis reaction was stopped by heating to 80 ℃ to obtain enzymatically hydrolyzed soybean phospholipid.
(2) Preparing a culture medium according to the following raw materials in percentage by weight: 30g of enzymolysis soybean phospholipid, 10g of molasses, 1g of monoammonium phosphate, 1.6g of urea, 1g of monopotassium phosphate, 0.17g of vitamin A, 20.1g of vitamin B, 0.2g of vitamin E, 0.13g of folic acid, 0.2g of EDTA-iron, 0.1g of EDTA-manganese, 0.001g of EDTA-copper and 0.1g of EDTA-zinc, 2g of sodium chloride and 53.5g of water.
(3) Inoculating 2g of bacillus subtilis seed solution into a culture medium, adjusting the pH value to be 6.0-6.5, and culturing for 32 hours to obtain 100g of microbial agent for enzymolysis of soybean phospholipid.
Example 5
(1) To 30g of the soybean phospholipid solution prepared in example 1, 0.18g of catalase, 0.09g of pepsin, and 0.03g of carboxypeptidase B were added, the pH was adjusted to 7.0, the enzymatic hydrolysis temperature was 35 ℃, and after 48 hours of enzymatic hydrolysis, the enzymatic hydrolysis reaction was stopped by heating to 80 ℃ to obtain enzymatically hydrolyzed soybean phospholipid.
(2) Preparing a culture medium according to the following raw materials in percentage by weight: 25g of enzymolysis soybean phospholipid, 15g of molasses, 1g of monoammonium phosphate, 1.6g of urea, 1g of monopotassium phosphate, 0.17g of vitamin A, 20.1g of vitamin B, 0.2g of vitamin E, 0.13g of folic acid, 0.2g of EDTA-iron, 0.1g of EDTA-manganese, 0.001g of EDTA-copper, 0.1g of EDTA-zinc, 1.5g of sodium chloride and 54g of water.
(3) Inoculating 1.5g of bacillus subtilis seed solution into a culture medium, adjusting the pH value to be 6.0-6.5, and culturing for 40 hours to obtain 100g of microbial agent for enzymolysis of soybean phospholipid.
Comparative example 1
(1) Preparing a culture medium according to the following raw materials in percentage by weight: 25g of soybean lecithin liquid prepared in example 1, 15g of molasses, 1g of monoammonium phosphate, 1.6g of urea, 1g of monopotassium phosphate, 0.17g of vitamin A, 20.1g of vitamin B, 0.2g of vitamin E and 0.13g of folic acid, 0.2g of EDTA-iron, 0.1g of EDTA-manganese, 0.001g of EDTA-copper and 0.1g of EDTA-zinc, 1.5g of sodium chloride and 54g of water.
(2) Inoculating 1.5g of Bacillus subtilis seed solution into culture medium, adjusting pH to 6.0-6.5, and culturing for 40 hr to obtain 100g of microbial agent.
Comparative example 2
(1) The components according to the following mass concentrations: preparing 100g of culture medium from 4g/L of beef extract, 9g/L of peptone and 4g/L of sodium chloride; inoculating 1.5g of Bacillus subtilis seed solution into a culture medium, adjusting pH to 6.0-6.5, and culturing for 40 hours to obtain Bacillus subtilis fermentation liquid.
(2) To 30g of the soybean phospholipid solution prepared in example 1, 0.18g of catalase, 0.09g of pepsin, and 0.03g of carboxypeptidase B were added, the pH was adjusted to 7.0, the enzymatic hydrolysis temperature was 30 ℃, and after 48 hours of enzymatic hydrolysis, the enzymatic hydrolysis reaction was stopped by heating to 80 ℃ to obtain enzymatically hydrolyzed soybean phospholipid.
(3) Preparing 100g of microbial agent for enzymolysis of soybean lecithin, 25g of enzymolysis soybean lecithin, 5g of bacillus subtilis fermentation liquor, 15g of molasses, 1g of monoammonium phosphate, 1.6g of urea, 1g of monopotassium phosphate, 0.17g of vitamin A, 20.1g of vitamin B, 0.2g of vitamin E, 0.13g of folic acid, 0.2g of EDTA-iron, 0.1g of EDTA-manganese, 0.001g of EDTA-copper, 0.1g of EDTA-zinc and 50.5g of water.
Test examples
With cantaloupe as the subject, 4 treatments were set, each treatment being repeated 3 times. Treatment 1(T1) was the microbial agent prepared by the method of comparative example 1, and the microbial agent was mixed with water at a weight ratio of 1:100 and then irrigated with melon seedlings at a dose of 150kg/ha, treatment 2(T2) was the enzymatic soybean lecithin microbial agent prepared by the method of comparative example 2, and the microbial agent was mixed with water at a weight ratio of 1:100 and then irrigated with melon seedlings at a dose of 150kg/ha, and treatment 3(T3) was the enzymatic soybean lecithin microbial agent prepared by the method of example 5, and the microbial agent was mixed with water at a weight ratio of 1:100 and then irrigated with melon seedlings at a dose of 150kg/ha, and the same amount of clear water was applied as control treatment (CK). The melon management is daily management, after the melon grows for 60 days, the influence of the enzymolysis soybean phospholipid microbial inoculum on the growth characteristics of the melon in the vine extending period is inspected through measuring chlorophyll, stem length and diameter of the melon, and the obtained result is shown in table 1.
TABLE 1 Effect of microbial Agents on muskmelon growth characteristics in the Finishes stage
As can be seen from table 1, the microbial agent for enzymatically hydrolyzing soybean phospholipid prepared by the method in example 5 can increase the chlorophyll content in the leaves of melon to the maximum, thereby facilitating photosynthesis of melon and promoting growth and development; the longest of the melon vine,
The stem thickness was largest. The comparative example 1 is to use soybean phospholipid to replace enzymolysis soybean phospholipid to prepare the microbial agent, the comparative example 2 is to use the traditional method to expand bacillus subtilis, and the obtained fermentation liquor is prepared into the microbial agent with the raw materials of enzymolysis soybean phospholipid, molasses and the like. After acting on the melons respectively, it can be seen that: the microbial agent for enzymolysis of soybean phospholipid can effectively promote the growth and development of crops, and the growth promoting effect is better than that of comparative examples 1 and 2. The bacillus subtilis is fermented in a culture medium prepared from 20-30% of enzymolysis soybean phospholipid, 10-20% of molasses, 1% of monoammonium phosphate, 1.6% of urea, 1% of monopotassium phosphate, 0.6% of vitamin, 0.401% of trace elements, 1-2% of sodium chloride and the balance water, so that the obtained effective viable bacteria are large in quantity, macromolecular substances can be decomposed into small molecular substances which are easy to absorb by plants, the growth of the plants is promoted, and the utilization rate of the bacillus subtilis is improved.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (10)
1. The microbial agent for enzymolysis of soybean phospholipid is characterized by being prepared by the following steps:
(1) adding catalase, pepsin and carboxypeptidase B into the soybean phospholipid liquid, adjusting the pH value to 7.0, performing enzymolysis for 48 hours, heating to 80 ℃, and stopping enzymolysis reaction to obtain enzymolysis soybean phospholipid;
(2) preparing a culture medium according to the following raw materials in percentage by weight: 20-30% of enzymolysis soybean phospholipid, 10-20% of molasses, 1% of monoammonium phosphate, 1.6% of urea, 1% of monopotassium phosphate, 0.6% of vitamin, 0.4% of trace elements, 1-2% of sodium chloride and the balance of water;
(3) inoculating the bacillus subtilis seed solution into a culture medium according to the inoculation amount of 1-2%, adjusting the pH to 6.0-6.5, and culturing for 32-48 hours to obtain the microbial agent for enzymolysis of soybean lecithin.
2. The microbial agent for enzymatic hydrolysis of soybean phospholipid as defined in claim 1, wherein in the step (1), the soybean phospholipid solution is prepared by the following method:
1) dehydrating the soybean oil residue to obtain crude phospholipid;
2) adding acetone with equal mass into the crude phospholipid, stirring, performing solid-liquid separation after stirring is finished, and recovering the acetone to obtain crude phospholipid powder;
3) adding 30% hydrogen peroxide of 1% of the weight of the material into the crude phospholipid powder, fully stirring and decoloring, and steaming out the hydrogen peroxide to obtain refined soybean phospholipid;
4) dissolving refined soybean phospholipid in vegetable oil to obtain soybean phospholipid solution.
3. The microbial agent for enzymolysis of soybean phospholipid as claimed in claim 2, wherein the addition amount of refined soybean phospholipid and vegetable oil is 10-5: 1; the vegetable oil is one or more of soybean oil, corn oil, rapeseed oil and peanut oil.
4. The microbial agent for the enzymatic hydrolysis of soybean phospholipid as set forth in claim 1, wherein in the step (1), the catalase is added in an amount of 0.6% by weight, the pepsin is added in an amount of 0.3% by weight, and the carboxypeptidase B is added in an amount of 0.1% by weight; the enzymolysis temperature is 25-35 ℃.
5. The microbial agent for enzymolysis of soybean phospholipid as claimed in claim 1, wherein in the step (2), the molasses is cane molasses, the brix of the molasses is 80-90 Bx, and the sugar content of the molasses is more than 50%.
6. The microbial agent for enzymolysis of soybean phospholipid as claimed in claim 1, wherein in step (2), the vitamins include vitamin A, vitamin B2, vitamin E and folic acid, and the mass ratio of vitamin A, vitamin B2, vitamin E and folic acid is 1.7: 1: 2: 1.3.
7. the microbial agent for enzymolysis of soybean phospholipid as claimed in claim 1, wherein in the step (2), the trace elements comprise EDTA-iron, EDTA-manganese, EDTA-copper and EDTA-zinc, and the mass ratio of the EDTA-iron, the EDTA-manganese, the EDTA-copper and the EDTA-zinc is 2: 1: 0.01: 1.
8. the microbial agent for enzymatic hydrolysis of soybean phospholipid as defined in claim 1, wherein in the step (3), the number of effective viable bacteria in the microbial agent for enzymatic hydrolysis of soybean phospholipid is not less than 20.0X 109cfu/mL。
9. The use of the enzymatically hydrolyzed soybean phospholipid microbial agent of claims 1-8 for promoting crop growth.
10. The use according to claim 9, wherein the crop growth promotion is the promotion of increased chlorophyll content in a crop and the promotion of growth of crop shoots.
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