CN104480089B - The zytase of mutation and its application in L lysine products - Google Patents
The zytase of mutation and its application in L lysine products Download PDFInfo
- Publication number
- CN104480089B CN104480089B CN201410777733.6A CN201410777733A CN104480089B CN 104480089 B CN104480089 B CN 104480089B CN 201410777733 A CN201410777733 A CN 201410777733A CN 104480089 B CN104480089 B CN 104480089B
- Authority
- CN
- China
- Prior art keywords
- zytase
- acquisition
- coating
- product containing
- present
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 230000035772 mutation Effects 0.000 title claims abstract description 15
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 title abstract description 25
- 239000004472 Lysine Substances 0.000 title abstract description 24
- 239000011248 coating agent Substances 0.000 claims description 53
- 238000000576 coating method Methods 0.000 claims description 53
- 239000002245 particle Substances 0.000 claims description 47
- 238000000034 method Methods 0.000 claims description 34
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 31
- 235000013923 monosodium glutamate Nutrition 0.000 claims description 31
- 229940073490 sodium glutamate Drugs 0.000 claims description 31
- 230000000694 effects Effects 0.000 claims description 28
- 239000008187 granular material Substances 0.000 claims description 24
- 239000000203 mixture Substances 0.000 claims description 24
- 239000000945 filler Substances 0.000 claims description 22
- 239000011122 softwood Substances 0.000 claims description 18
- 230000001070 adhesive effect Effects 0.000 claims description 14
- 108090000623 proteins and genes Proteins 0.000 claims description 14
- 239000000853 adhesive Substances 0.000 claims description 13
- 238000001035 drying Methods 0.000 claims description 10
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 8
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 8
- 239000011230 binding agent Substances 0.000 claims description 8
- 238000007873 sieving Methods 0.000 claims description 8
- 238000000855 fermentation Methods 0.000 claims description 5
- 230000004151 fermentation Effects 0.000 claims description 5
- 239000002773 nucleotide Substances 0.000 claims description 5
- 125000003729 nucleotide group Chemical group 0.000 claims description 5
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical class CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 claims description 4
- 230000003248 secreting effect Effects 0.000 claims description 4
- 241000894006 Bacteria Species 0.000 claims description 3
- 239000003925 fat Substances 0.000 claims 2
- 238000006243 chemical reaction Methods 0.000 claims 1
- 238000001890 transfection Methods 0.000 claims 1
- 102200012961 rs121918637 Human genes 0.000 abstract description 5
- 239000000047 product Substances 0.000 description 53
- 229960003646 lysine Drugs 0.000 description 22
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 description 15
- 238000002360 preparation method Methods 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 11
- 102000004190 Enzymes Human genes 0.000 description 11
- 238000006731 degradation reaction Methods 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 238000012216 screening Methods 0.000 description 8
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 7
- 235000014590 basal diet Nutrition 0.000 description 7
- 229920001353 Dextrin Polymers 0.000 description 6
- 239000004375 Dextrin Substances 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 6
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 6
- 235000019425 dextrin Nutrition 0.000 description 6
- 238000005469 granulation Methods 0.000 description 6
- 230000003179 granulation Effects 0.000 description 6
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 6
- 239000008108 microcrystalline cellulose Substances 0.000 description 6
- 229940016286 microcrystalline cellulose Drugs 0.000 description 6
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 5
- 244000309466 calf Species 0.000 description 5
- 239000001768 carboxy methyl cellulose Substances 0.000 description 5
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 5
- 230000018044 dehydration Effects 0.000 description 5
- 238000006297 dehydration reaction Methods 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 239000000654 additive Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000000717 retained effect Effects 0.000 description 3
- 210000004767 rumen Anatomy 0.000 description 3
- 229910021653 sulphate ion Inorganic materials 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 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
- YYSWCHMLFJLLBJ-ZLUOBGJFSA-N Ala-Ala-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YYSWCHMLFJLLBJ-ZLUOBGJFSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 108010001682 Dextranase Proteins 0.000 description 2
- RGXXLQWXBFNXTG-CIUDSAMLSA-N Gln-Arg-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O RGXXLQWXBFNXTG-CIUDSAMLSA-N 0.000 description 2
- CKRUHITYRFNUKW-WDSKDSINSA-N Glu-Asn-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O CKRUHITYRFNUKW-WDSKDSINSA-N 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- BVHLGVCQOALMSV-JEDNCBNOSA-N L-lysine hydrochloride Chemical group Cl.NCCCC[C@H](N)C(O)=O BVHLGVCQOALMSV-JEDNCBNOSA-N 0.000 description 2
- 241000880493 Leptailurus serval Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 101000702488 Rattus norvegicus High affinity cationic amino acid transporter 1 Proteins 0.000 description 2
- BRKHVZNDAOMAHX-BIIVOSGPSA-N Ser-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N BRKHVZNDAOMAHX-BIIVOSGPSA-N 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000003674 animal food additive Substances 0.000 description 2
- 108010047857 aspartylglycine Proteins 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000005422 blasting Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 235000010445 lecithin Nutrition 0.000 description 2
- 239000000787 lecithin Substances 0.000 description 2
- 229940067606 lecithin Drugs 0.000 description 2
- 229960005337 lysine hydrochloride Drugs 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 235000011009 potassium phosphates Nutrition 0.000 description 2
- 238000005057 refrigeration Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000002023 wood Substances 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- DKJPOZOEBONHFS-ZLUOBGJFSA-N Ala-Ala-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O DKJPOZOEBONHFS-ZLUOBGJFSA-N 0.000 description 1
- XQJAFSDFQZPYCU-UWJYBYFXSA-N Ala-Asn-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N XQJAFSDFQZPYCU-UWJYBYFXSA-N 0.000 description 1
- ZIWWTZWAKYBUOB-CIUDSAMLSA-N Ala-Asp-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O ZIWWTZWAKYBUOB-CIUDSAMLSA-N 0.000 description 1
- PUBLUECXJRHTBK-ACZMJKKPSA-N Ala-Glu-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O PUBLUECXJRHTBK-ACZMJKKPSA-N 0.000 description 1
- CYBJZLQSUJEMAS-LFSVMHDDSA-N Ala-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](C)N)O CYBJZLQSUJEMAS-LFSVMHDDSA-N 0.000 description 1
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 1
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 1
- SYIFFFHSXBNPMC-UWJYBYFXSA-N Ala-Ser-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N SYIFFFHSXBNPMC-UWJYBYFXSA-N 0.000 description 1
- YKBHOXLMMPZPHQ-GMOBBJLQSA-N Arg-Ile-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O YKBHOXLMMPZPHQ-GMOBBJLQSA-N 0.000 description 1
- AMIQZQAAYGYKOP-FXQIFTODSA-N Arg-Ser-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O AMIQZQAAYGYKOP-FXQIFTODSA-N 0.000 description 1
- AIFHRTPABBBHKU-RCWTZXSCSA-N Arg-Thr-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O AIFHRTPABBBHKU-RCWTZXSCSA-N 0.000 description 1
- ZZXMOQIUIJJOKZ-ZLUOBGJFSA-N Asn-Asn-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(N)=O ZZXMOQIUIJJOKZ-ZLUOBGJFSA-N 0.000 description 1
- PIWWUBYJNONVTJ-ZLUOBGJFSA-N Asn-Asp-Asn Chemical compound C([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)C(=O)N PIWWUBYJNONVTJ-ZLUOBGJFSA-N 0.000 description 1
- VYLVOMUVLMGCRF-ZLUOBGJFSA-N Asn-Asp-Ser Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O VYLVOMUVLMGCRF-ZLUOBGJFSA-N 0.000 description 1
- OGMDXNFGPOPZTK-GUBZILKMSA-N Asn-Glu-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC(=O)N)N OGMDXNFGPOPZTK-GUBZILKMSA-N 0.000 description 1
- CTQIOCMSIJATNX-WHFBIAKZSA-N Asn-Gly-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C)C(O)=O CTQIOCMSIJATNX-WHFBIAKZSA-N 0.000 description 1
- HPNDKUOLNRVRAY-BIIVOSGPSA-N Asn-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N)C(=O)O HPNDKUOLNRVRAY-BIIVOSGPSA-N 0.000 description 1
- HNXWVVHIGTZTBO-LKXGYXEUSA-N Asn-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O HNXWVVHIGTZTBO-LKXGYXEUSA-N 0.000 description 1
- ZVTDYGWRRPMFCL-WFBYXXMGSA-N Asp-Ala-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC(=O)O)N ZVTDYGWRRPMFCL-WFBYXXMGSA-N 0.000 description 1
- QXHVOUSPVAWEMX-ZLUOBGJFSA-N Asp-Asp-Ser Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O QXHVOUSPVAWEMX-ZLUOBGJFSA-N 0.000 description 1
- KHGPWGKPYHPOIK-QWRGUYRKSA-N Asp-Gly-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O KHGPWGKPYHPOIK-QWRGUYRKSA-N 0.000 description 1
- DWOGMPWRQQWPPF-GUBZILKMSA-N Asp-Leu-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O DWOGMPWRQQWPPF-GUBZILKMSA-N 0.000 description 1
- YFGUZQQCSDZRBN-DCAQKATOSA-N Asp-Pro-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O YFGUZQQCSDZRBN-DCAQKATOSA-N 0.000 description 1
- MJJIHRWNWSQTOI-VEVYYDQMSA-N Asp-Thr-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O MJJIHRWNWSQTOI-VEVYYDQMSA-N 0.000 description 1
- IWLZBRTUIVXZJD-OLHMAJIHSA-N Asp-Thr-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O IWLZBRTUIVXZJD-OLHMAJIHSA-N 0.000 description 1
- FIRWLDUOFOULCA-XIRDDKMYSA-N Asp-Trp-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)O)N FIRWLDUOFOULCA-XIRDDKMYSA-N 0.000 description 1
- RKXVTTIQNKPCHU-KKHAAJSZSA-N Asp-Val-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC(O)=O RKXVTTIQNKPCHU-KKHAAJSZSA-N 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- KZKBJEUWNMQTLV-XDTLVQLUSA-N Gln-Ala-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KZKBJEUWNMQTLV-XDTLVQLUSA-N 0.000 description 1
- ODBLJLZVLAWVMS-GUBZILKMSA-N Gln-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)N)N ODBLJLZVLAWVMS-GUBZILKMSA-N 0.000 description 1
- SDSMVVSHLAAOJL-UKJIMTQDSA-N Gln-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCC(=O)N)N SDSMVVSHLAAOJL-UKJIMTQDSA-N 0.000 description 1
- ATRHMOJQJWPVBQ-DRZSPHRISA-N Glu-Ala-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ATRHMOJQJWPVBQ-DRZSPHRISA-N 0.000 description 1
- WVTIBGWZUMJBFY-GUBZILKMSA-N Glu-His-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O WVTIBGWZUMJBFY-GUBZILKMSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- UGVQELHRNUDMAA-BYPYZUCNSA-N Gly-Ala-Gly Chemical compound [NH3+]CC(=O)N[C@@H](C)C(=O)NCC([O-])=O UGVQELHRNUDMAA-BYPYZUCNSA-N 0.000 description 1
- JRDYDYXZKFNNRQ-XPUUQOCRSA-N Gly-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN JRDYDYXZKFNNRQ-XPUUQOCRSA-N 0.000 description 1
- LCNXZQROPKFGQK-WHFBIAKZSA-N Gly-Asp-Ser Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O LCNXZQROPKFGQK-WHFBIAKZSA-N 0.000 description 1
- QPDUVFSVVAOUHE-XVKPBYJWSA-N Gly-Gln-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCC(N)=O)NC(=O)CN)C(O)=O QPDUVFSVVAOUHE-XVKPBYJWSA-N 0.000 description 1
- BUEFQXUHTUZXHR-LURJTMIESA-N Gly-Gly-Pro zwitterion Chemical compound NCC(=O)NCC(=O)N1CCC[C@H]1C(O)=O BUEFQXUHTUZXHR-LURJTMIESA-N 0.000 description 1
- INLIXXRWNUKVCF-JTQLQIEISA-N Gly-Gly-Tyr Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 INLIXXRWNUKVCF-JTQLQIEISA-N 0.000 description 1
- FSPVILZGHUJOHS-QWRGUYRKSA-N Gly-His-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CNC=N1 FSPVILZGHUJOHS-QWRGUYRKSA-N 0.000 description 1
- YTSVAIMKVLZUDU-YUMQZZPRSA-N Gly-Leu-Asp Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O YTSVAIMKVLZUDU-YUMQZZPRSA-N 0.000 description 1
- TWTPDFFBLQEBOE-IUCAKERBSA-N Gly-Leu-Gln Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O TWTPDFFBLQEBOE-IUCAKERBSA-N 0.000 description 1
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 1
- LHYJCVCQPWRMKZ-WEDXCCLWSA-N Gly-Leu-Thr Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LHYJCVCQPWRMKZ-WEDXCCLWSA-N 0.000 description 1
- LOEANKRDMMVOGZ-YUMQZZPRSA-N Gly-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)N[C@@H](CC(O)=O)C(O)=O LOEANKRDMMVOGZ-YUMQZZPRSA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- ABPRMMYHROQBLY-NKWVEPMBSA-N Gly-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)CN)C(=O)O ABPRMMYHROQBLY-NKWVEPMBSA-N 0.000 description 1
- CQMFNTVQVLQRLT-JHEQGTHGSA-N Gly-Thr-Gln Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O CQMFNTVQVLQRLT-JHEQGTHGSA-N 0.000 description 1
- IHDKKJVBLGXLEL-STQMWFEESA-N Gly-Tyr-Met Chemical compound CSCC[C@H](NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)CN)C(O)=O IHDKKJVBLGXLEL-STQMWFEESA-N 0.000 description 1
- RYAOJUMWLWUGNW-QMMMGPOBSA-N Gly-Val-Gly Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O RYAOJUMWLWUGNW-QMMMGPOBSA-N 0.000 description 1
- HIAHVKLTHNOENC-HGNGGELXSA-N His-Glu-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O HIAHVKLTHNOENC-HGNGGELXSA-N 0.000 description 1
- FZKFYOXDVWDELO-KBPBESRZSA-N His-Gly-Tyr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O FZKFYOXDVWDELO-KBPBESRZSA-N 0.000 description 1
- ORERHHPZDDEMSC-VGDYDELISA-N His-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N ORERHHPZDDEMSC-VGDYDELISA-N 0.000 description 1
- FCPSGEVYIVXPPO-QTKMDUPCSA-N His-Thr-Arg Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FCPSGEVYIVXPPO-QTKMDUPCSA-N 0.000 description 1
- XENGULNPUDGALZ-ZPFDUUQYSA-N Ile-Asn-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(C)C)C(=O)O)N XENGULNPUDGALZ-ZPFDUUQYSA-N 0.000 description 1
- GECLQMBTZCPAFY-PEFMBERDSA-N Ile-Gln-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N GECLQMBTZCPAFY-PEFMBERDSA-N 0.000 description 1
- NZOCIWKZUVUNDW-ZKWXMUAHSA-N Ile-Gly-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O NZOCIWKZUVUNDW-ZKWXMUAHSA-N 0.000 description 1
- LBRCLQMZAHRTLV-ZKWXMUAHSA-N Ile-Gly-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O LBRCLQMZAHRTLV-ZKWXMUAHSA-N 0.000 description 1
- CCYGNFBYUNHFSC-MGHWNKPDSA-N Ile-His-Phe Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O CCYGNFBYUNHFSC-MGHWNKPDSA-N 0.000 description 1
- RMNMUUCYTMLWNA-ZPFDUUQYSA-N Ile-Lys-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)O)C(=O)O)N RMNMUUCYTMLWNA-ZPFDUUQYSA-N 0.000 description 1
- LZDNBBYBDGBADK-UHFFFAOYSA-N L-valyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C(C)C)C(O)=O)=CNC2=C1 LZDNBBYBDGBADK-UHFFFAOYSA-N 0.000 description 1
- KKXDHFKZWKLYGB-GUBZILKMSA-N Leu-Asn-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N KKXDHFKZWKLYGB-GUBZILKMSA-N 0.000 description 1
- IASQBRJGRVXNJI-YUMQZZPRSA-N Leu-Cys-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)NCC(O)=O IASQBRJGRVXNJI-YUMQZZPRSA-N 0.000 description 1
- KEVYYIMVELOXCT-KBPBESRZSA-N Leu-Gly-Phe Chemical compound CC(C)C[C@H]([NH3+])C(=O)NCC(=O)N[C@H](C([O-])=O)CC1=CC=CC=C1 KEVYYIMVELOXCT-KBPBESRZSA-N 0.000 description 1
- PTRKPHUGYULXPU-KKUMJFAQSA-N Leu-Phe-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O PTRKPHUGYULXPU-KKUMJFAQSA-N 0.000 description 1
- YUTNOGOMBNYPFH-XUXIUFHCSA-N Leu-Pro-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YUTNOGOMBNYPFH-XUXIUFHCSA-N 0.000 description 1
- IZPVWNSAVUQBGP-CIUDSAMLSA-N Leu-Ser-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IZPVWNSAVUQBGP-CIUDSAMLSA-N 0.000 description 1
- SBANPBVRHYIMRR-UHFFFAOYSA-N Leu-Ser-Pro Natural products CC(C)CC(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O SBANPBVRHYIMRR-UHFFFAOYSA-N 0.000 description 1
- AEDWWMMHUGYIFD-HJGDQZAQSA-N Leu-Thr-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O AEDWWMMHUGYIFD-HJGDQZAQSA-N 0.000 description 1
- LJBVRCDPWOJOEK-PPCPHDFISA-N Leu-Thr-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LJBVRCDPWOJOEK-PPCPHDFISA-N 0.000 description 1
- CNWDWAMPKVYJJB-NUTKFTJISA-N Leu-Trp-Ala Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](N)CC(C)C)C(=O)N[C@@H](C)C(O)=O)=CNC2=C1 CNWDWAMPKVYJJB-NUTKFTJISA-N 0.000 description 1
- FZIJIFCXUCZHOL-CIUDSAMLSA-N Lys-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN FZIJIFCXUCZHOL-CIUDSAMLSA-N 0.000 description 1
- RIPJMCFGQHGHNP-RHYQMDGZSA-N Lys-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCCCN)N)O RIPJMCFGQHGHNP-RHYQMDGZSA-N 0.000 description 1
- FWAHLGXNBLWIKB-NAKRPEOUSA-N Met-Ile-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCSC FWAHLGXNBLWIKB-NAKRPEOUSA-N 0.000 description 1
- AWGBEIYZPAXXSX-RWMBFGLXSA-N Met-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCSC)N AWGBEIYZPAXXSX-RWMBFGLXSA-N 0.000 description 1
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 1
- 108010066427 N-valyltryptophan Proteins 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- IILUKIJNFMUBNF-IHRRRGAJSA-N Phe-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O IILUKIJNFMUBNF-IHRRRGAJSA-N 0.000 description 1
- YYKZDTVQHTUKDW-RYUDHWBXSA-N Phe-Gly-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N YYKZDTVQHTUKDW-RYUDHWBXSA-N 0.000 description 1
- WEMYTDDMDBLPMI-DKIMLUQUSA-N Phe-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N WEMYTDDMDBLPMI-DKIMLUQUSA-N 0.000 description 1
- METZZBCMDXHFMK-BZSNNMDCSA-N Phe-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N METZZBCMDXHFMK-BZSNNMDCSA-N 0.000 description 1
- YOFKMVUAZGPFCF-IHRRRGAJSA-N Phe-Met-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(O)=O YOFKMVUAZGPFCF-IHRRRGAJSA-N 0.000 description 1
- QSWKNJAPHQDAAS-MELADBBJSA-N Phe-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O QSWKNJAPHQDAAS-MELADBBJSA-N 0.000 description 1
- QTDBZORPVYTRJU-KKXDTOCCSA-N Phe-Tyr-Ala Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O QTDBZORPVYTRJU-KKXDTOCCSA-N 0.000 description 1
- 241000235648 Pichia Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- UPJGUQPLYWTISV-GUBZILKMSA-N Pro-Gln-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O UPJGUQPLYWTISV-GUBZILKMSA-N 0.000 description 1
- MGDFPGCFVJFITQ-CIUDSAMLSA-N Pro-Glu-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MGDFPGCFVJFITQ-CIUDSAMLSA-N 0.000 description 1
- VYWNORHENYEQDW-YUMQZZPRSA-N Pro-Gly-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 VYWNORHENYEQDW-YUMQZZPRSA-N 0.000 description 1
- PEYNRYREGPAOAK-LSJOCFKGSA-N Pro-His-Ala Chemical compound C([C@@H](C(=O)N[C@@H](C)C([O-])=O)NC(=O)[C@H]1[NH2+]CCC1)C1=CN=CN1 PEYNRYREGPAOAK-LSJOCFKGSA-N 0.000 description 1
- XFFIGWGYMUFCCQ-ULQDDVLXSA-N Pro-His-Tyr Chemical compound C1=CC(O)=CC=C1C[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)[C@H]1[NH2+]CCC1)CC1=CN=CN1 XFFIGWGYMUFCCQ-ULQDDVLXSA-N 0.000 description 1
- LNOWDSPAYBWJOR-PEDHHIEDSA-N Pro-Ile-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LNOWDSPAYBWJOR-PEDHHIEDSA-N 0.000 description 1
- HFNPOYOKIPGAEI-SRVKXCTJSA-N Pro-Leu-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 HFNPOYOKIPGAEI-SRVKXCTJSA-N 0.000 description 1
- SUENWIFTSTWUKD-AVGNSLFASA-N Pro-Leu-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O SUENWIFTSTWUKD-AVGNSLFASA-N 0.000 description 1
- GZNYIXWOIUFLGO-ZJDVBMNYSA-N Pro-Thr-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZNYIXWOIUFLGO-ZJDVBMNYSA-N 0.000 description 1
- CXGLFEOYCJFKPR-RCWTZXSCSA-N Pro-Thr-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O CXGLFEOYCJFKPR-RCWTZXSCSA-N 0.000 description 1
- MEVPFKSOSFRIGY-LURJTMIESA-N S(=O)(=O)(O)OCC[C@H](NCCC)C(=O)O Chemical compound S(=O)(=O)(O)OCC[C@H](NCCC)C(=O)O MEVPFKSOSFRIGY-LURJTMIESA-N 0.000 description 1
- HRNQLKCLPVKZNE-CIUDSAMLSA-N Ser-Ala-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O HRNQLKCLPVKZNE-CIUDSAMLSA-N 0.000 description 1
- IDQFQFVEWMWRQQ-DLOVCJGASA-N Ser-Ala-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O IDQFQFVEWMWRQQ-DLOVCJGASA-N 0.000 description 1
- QGMLKFGTGXWAHF-IHRRRGAJSA-N Ser-Arg-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QGMLKFGTGXWAHF-IHRRRGAJSA-N 0.000 description 1
- QPFJSHSJFIYDJZ-GHCJXIJMSA-N Ser-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO QPFJSHSJFIYDJZ-GHCJXIJMSA-N 0.000 description 1
- DJACUBDEDBZKLQ-KBIXCLLPSA-N Ser-Ile-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O DJACUBDEDBZKLQ-KBIXCLLPSA-N 0.000 description 1
- IXZHZUGGKLRHJD-DCAQKATOSA-N Ser-Leu-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O IXZHZUGGKLRHJD-DCAQKATOSA-N 0.000 description 1
- NNFMANHDYSVNIO-DCAQKATOSA-N Ser-Lys-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NNFMANHDYSVNIO-DCAQKATOSA-N 0.000 description 1
- LRZLZIUXQBIWTB-KATARQTJSA-N Ser-Lys-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LRZLZIUXQBIWTB-KATARQTJSA-N 0.000 description 1
- QMCDMHWAKMUGJE-IHRRRGAJSA-N Ser-Phe-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O QMCDMHWAKMUGJE-IHRRRGAJSA-N 0.000 description 1
- FKYWFUYPVKLJLP-DCAQKATOSA-N Ser-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FKYWFUYPVKLJLP-DCAQKATOSA-N 0.000 description 1
- YLXAMFZYJTZXFH-OLHMAJIHSA-N Thr-Asn-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O YLXAMFZYJTZXFH-OLHMAJIHSA-N 0.000 description 1
- DGOJNGCGEYOBKN-BWBBJGPYSA-N Thr-Cys-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CS)C(=O)O)N)O DGOJNGCGEYOBKN-BWBBJGPYSA-N 0.000 description 1
- VULNJDORNLBPNG-SWRJLBSHSA-N Thr-Glu-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O VULNJDORNLBPNG-SWRJLBSHSA-N 0.000 description 1
- KCRQEJSKXAIULJ-FJXKBIBVSA-N Thr-Gly-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O KCRQEJSKXAIULJ-FJXKBIBVSA-N 0.000 description 1
- STUAPCLEDMKXKL-LKXGYXEUSA-N Thr-Ser-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O STUAPCLEDMKXKL-LKXGYXEUSA-N 0.000 description 1
- NBIIPOKZPUGATB-BWBBJGPYSA-N Thr-Ser-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N)O NBIIPOKZPUGATB-BWBBJGPYSA-N 0.000 description 1
- AHERARIZBPOMNU-KATARQTJSA-N Thr-Ser-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O AHERARIZBPOMNU-KATARQTJSA-N 0.000 description 1
- BDWDMRSGCXEDMR-WFBYXXMGSA-N Trp-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N BDWDMRSGCXEDMR-WFBYXXMGSA-N 0.000 description 1
- CZWIHKFGHICAJX-BPUTZDHNSA-N Trp-Glu-Glu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 CZWIHKFGHICAJX-BPUTZDHNSA-N 0.000 description 1
- RWAYYYOZMHMEGD-XIRDDKMYSA-N Trp-Leu-Ser Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O)=CNC2=C1 RWAYYYOZMHMEGD-XIRDDKMYSA-N 0.000 description 1
- CRCHQCUINSOGFD-JBACZVJFSA-N Trp-Tyr-Glu Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CC=C(C=C3)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N CRCHQCUINSOGFD-JBACZVJFSA-N 0.000 description 1
- LGEYOIQBBIPHQN-UWJYBYFXSA-N Tyr-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 LGEYOIQBBIPHQN-UWJYBYFXSA-N 0.000 description 1
- QUILOGWWLXMSAT-IHRRRGAJSA-N Tyr-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QUILOGWWLXMSAT-IHRRRGAJSA-N 0.000 description 1
- UNUZEBFXGWVAOP-DZKIICNBSA-N Tyr-Glu-Val Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O UNUZEBFXGWVAOP-DZKIICNBSA-N 0.000 description 1
- NSGZILIDHCIZAM-KKUMJFAQSA-N Tyr-Leu-Ser Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N NSGZILIDHCIZAM-KKUMJFAQSA-N 0.000 description 1
- XUIOBCQESNDTDE-FQPOAREZSA-N Tyr-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N)O XUIOBCQESNDTDE-FQPOAREZSA-N 0.000 description 1
- RGJZPXFZIUUQDN-BPNCWPANSA-N Tyr-Val-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O RGJZPXFZIUUQDN-BPNCWPANSA-N 0.000 description 1
- SLLKXDSRVAOREO-KZVJFYERSA-N Val-Ala-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](C(C)C)N)O SLLKXDSRVAOREO-KZVJFYERSA-N 0.000 description 1
- UUYCNAXCCDNULB-QXEWZRGKSA-N Val-Arg-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(N)=O)C(O)=O UUYCNAXCCDNULB-QXEWZRGKSA-N 0.000 description 1
- CVUDMNSZAIZFAE-TUAOUCFPSA-N Val-Arg-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(=O)O)N CVUDMNSZAIZFAE-TUAOUCFPSA-N 0.000 description 1
- CVUDMNSZAIZFAE-UHFFFAOYSA-N Val-Arg-Pro Natural products NC(N)=NCCCC(NC(=O)C(N)C(C)C)C(=O)N1CCCC1C(O)=O CVUDMNSZAIZFAE-UHFFFAOYSA-N 0.000 description 1
- QPZMOUMNTGTEFR-ZKWXMUAHSA-N Val-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N QPZMOUMNTGTEFR-ZKWXMUAHSA-N 0.000 description 1
- PVPAOIGJYHVWBT-KKHAAJSZSA-N Val-Asn-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N)O PVPAOIGJYHVWBT-KKHAAJSZSA-N 0.000 description 1
- TZVUSFMQWPWHON-NHCYSSNCSA-N Val-Asp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C(C)C)N TZVUSFMQWPWHON-NHCYSSNCSA-N 0.000 description 1
- AGKDVLSDNSTLFA-UMNHJUIQSA-N Val-Gln-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N AGKDVLSDNSTLFA-UMNHJUIQSA-N 0.000 description 1
- AAOPYWQQBXHINJ-DZKIICNBSA-N Val-Gln-Tyr Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N AAOPYWQQBXHINJ-DZKIICNBSA-N 0.000 description 1
- PMDOQZFYGWZSTK-LSJOCFKGSA-N Val-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)C(C)C PMDOQZFYGWZSTK-LSJOCFKGSA-N 0.000 description 1
- APQIVBCUIUDSMB-OSUNSFLBSA-N Val-Ile-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C(C)C)N APQIVBCUIUDSMB-OSUNSFLBSA-N 0.000 description 1
- AGXGCFSECFQMKB-NHCYSSNCSA-N Val-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N AGXGCFSECFQMKB-NHCYSSNCSA-N 0.000 description 1
- ZRSZTKTVPNSUNA-IHRRRGAJSA-N Val-Lys-Leu Chemical compound CC(C)C[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)C(C)C)C(O)=O ZRSZTKTVPNSUNA-IHRRRGAJSA-N 0.000 description 1
- DEGUERSKQBRZMZ-FXQIFTODSA-N Val-Ser-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O DEGUERSKQBRZMZ-FXQIFTODSA-N 0.000 description 1
- KSFXWENSJABBFI-ZKWXMUAHSA-N Val-Ser-Asn Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KSFXWENSJABBFI-ZKWXMUAHSA-N 0.000 description 1
- NZYNRRGJJVSSTJ-GUBZILKMSA-N Val-Ser-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NZYNRRGJJVSSTJ-GUBZILKMSA-N 0.000 description 1
- HTONZBWRYUKUKC-RCWTZXSCSA-N Val-Thr-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O HTONZBWRYUKUKC-RCWTZXSCSA-N 0.000 description 1
- SUGRIIAOLCDLBD-ZOBUZTSGSA-N Val-Trp-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(=O)O)C(=O)O)N SUGRIIAOLCDLBD-ZOBUZTSGSA-N 0.000 description 1
- RFZFBOQPPFCOKG-BZSNNMDCSA-N Val-Trp-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCSC)C(=O)O)N RFZFBOQPPFCOKG-BZSNNMDCSA-N 0.000 description 1
- SNGZVXPRGLTKNU-RGMNGODLSA-N [Na].C(CC)N[C@@H](CCO)C(=O)O Chemical compound [Na].C(CC)N[C@@H](CCO)C(=O)O SNGZVXPRGLTKNU-RGMNGODLSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010045350 alanyl-tyrosyl-alanine Proteins 0.000 description 1
- 108010041407 alanylaspartic acid Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 108010011559 alanylphenylalanine Proteins 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 108010036533 arginylvaline Proteins 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 239000007767 bonding agent Substances 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000007771 core particle Substances 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 108010027668 glycyl-alanyl-valine Proteins 0.000 description 1
- 108010051307 glycyl-glycyl-proline Proteins 0.000 description 1
- 108010010096 glycyl-glycyl-tyrosine Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010084389 glycyltryptophan Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 108010083708 leucyl-aspartyl-valine Proteins 0.000 description 1
- 108010051673 leucyl-glycyl-phenylalanine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 108010009298 lysylglutamic acid Proteins 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229940029985 mineral supplement Drugs 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 108010025488 pinealon Proteins 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 108010079317 prolyl-tyrosine Proteins 0.000 description 1
- 108010029020 prolylglycine Proteins 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 108010071207 serylmethionine Proteins 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 108010033670 threonyl-aspartyl-tyrosine Proteins 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- IBIDRSSEHFLGSD-UHFFFAOYSA-N valinyl-arginine Natural products CC(C)C(N)C(=O)NC(C(O)=O)CCCN=C(N)N IBIDRSSEHFLGSD-UHFFFAOYSA-N 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 229920001221 xylan Polymers 0.000 description 1
- 150000004823 xylans Chemical class 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2477—Hemicellulases not provided in a preceding group
- C12N9/248—Xylanases
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Fodder In General (AREA)
Abstract
The invention provides the zytase of mutation, it includes mutation Arg45Ser.Other application present invention also offers it in the product for preparing the lysine containing L etc..
Description
Technical field
The invention belongs to enzyme field, specifically, contain L- the present invention relates to the zytase of new mutation and its preparing
Application in the product of lysine etc..
Background technology
1B is important amino acid starting material, can be used as flavouring, food, feed addictive, can also
As the effective or adjunct ingredient in health products, medicine, it is widely used in grocery trade, feed industry, pharmacy industry and other chemical works
In industry.Currently, the production of 1B mainly passes through the fermenting and producing of microorganism, can such as utilize corynebacteria production.
However, when lysine is as feed, because it belongs to small molecule nutriment, will soon be discharged into dynamic
In object, especially in huge uptake, the efficiency that animal absorbs have impact on.The method generally solved is to feed in multiple times on a small quantity
Animal is eaten, but this will increase the workload of animal feeding.Therefore, the present invention, which is proposed, directly uses fermenting lysine nutrient solution system
The method of standby coated lysine product, can effectively slow down the speed that lysine discharges in animal body.
The preceding Chinese patent 201210440083 of the present inventor describes a kind of product of coated lysine, main to use
In feed.On this basis, present inventor has performed in-depth study, unexpectedly in many aspects, including process aspect,
Composition（Including trace mineral supplement）In terms of aspect and enzyme transformation etc., obtain what can be improved coating and/or increase operation rate
The product of lysine.
Invention summary
The technical problem to be solved in the present invention is to provide the zytase of new mutation and its contain 1B preparing
Product in application.In addition, the present invention can also provide each branch invention obtained from product optimization, including rely containing L-
The feed addictive and product of propylhomoserin, and/or, the application of sodium glutamate in the preparation of the product containing 1B etc..
Preferably, in one aspect of the invention, the invention provides the zytase of mutation, it includes mutation
Arg45Ser。
It is preferred that the amino acid sequence of the zytase
（1）Such as SEQ ID No:Shown in 2；Or
（2）It is to SEQ ID No:2 lack, add and/or replace one or several（Preferably more than 7, more preferably not
More than 5,3 are such as no more than）Amino acid residue and the reservation SEQ ID No obtained:The sequence of 2 activity, and wherein include
It is mutated Arg45Ser.
On the other hand, the invention provides the gene for encoding foregoing zytase.
It is preferred that the nucleotide sequence of the gene such as SEQ ID No:Shown in 1.
On the other hand, the invention provides the carrier for including foregoing aspects of gene.
On the other hand, the invention provides strain（Especially secreting, expressing bacterium（Such as, yeast））, it includes foregoing aspect
Gene, or with foregoing aspects of carrier convert or transfect and obtain.
On the other hand, the invention provides the method for the foregoing aspects of zytase of fermenting and producing, it is included in fermentation
Under the conditions of cultivate foregoing aspects of strain, foregoing aspects of zytase is isolated from culture.
On the other hand, the product containing 1B is being prepared the invention provides foregoing aspects of zytase（Such as, raise
Feed additives）In purposes.
Preferably wherein, the coated layer that the product containing 1B includes coating slug particle and is wrapped on coating slug particle,
Wherein coated layer is rumen-undegraded fat, and coating slug particle includes foregoing aspects of zytase, lysine salt, filler and bonding
Agent and optional sodium glutamate.
Preferably wherein, the xylanase activity in every gram of product containing 1B described in claim 1 or 2 be 5 ~
15IU, more preferably 8 ~ 10IU, most preferably 9IU.
Preferably wherein, the method for preparation includes：
（1）Lysine salt and filler and optional sodium glutamate are well mixed, mixture is formed；
（2）To step（1）The binder solution containing zytase is added in the mixture of acquisition, is well mixed, is formed
Softwood；
（3）By step（2）The softwood sieving of acquisition（Such as, 30 mesh sieve）, form coarse granule；
（4）By step（3）The coarse granule ball blast of acquisition, forms circular micropill；
（5）Drying steps（4）The circular micropill of acquisition is simultaneously sieved, and forms coating slug particle；With,
（6）Optionally, rumen-undegraded fat is sprayed on step（5）On the coating slug particle of acquisition.
Detailed description of the invention
The technical problem to be solved in the present invention is to provide the zytase of new new mutation and its rely preparing containing L-
Application in the product of propylhomoserin.In addition, the present invention can also provide the preparation method of the feed addictive and the technique of optimization
Application in the feed addictive containing 1B is prepared, the product containing 1B, and/or, a small amount of sodium glutamate exists
The branches such as the application in the feed addictive containing 1B are prepared to invent.
One, term is explained
Herein, term has the implication that those skilled in the art routinely understands.For the sake of clarity, herein
Following term is illustrated.
Herein, term is " optional " has its dictionary justice, i.e., the object being accordingly defined is chosen or do not selected.Example
Such as, sodium glutamate is referred to containing or not contained containing optional sodium glutamate.And for example, optionally referred to containing zytase
Contain or do not contain zytase.
Herein, term " filler " and " adhesive " refer respectively to each serve as non-human animal's edible safety
Filling and the assistant agent of adhesive effect, it is preferable that " filler " and " adhesive " herein is food grade（That is people's edible safety
's）, it is referred to corresponding（People uses）The index of food.
Herein, term "comprising" or " comprising " are open statements, i.e., except list contain or comprise into
Outside part, can no longer contain other compositions or step, but be also not excluded for not influenceing containing other also the composition specifically listed or
Step plays the other compositions or step of function.Preferably, "comprising" herein or " comprising " are preferably " Consists of ".
Herein, as do not indicated plus especially, term " lysine " refers to 1B.
Two, the product containing 1B
The second of the present invention（1）Aspect, the invention provides the feed addictive containing 1B, it includes being coated with core
Particle and the coated layer being wrapped on coating slug particle, wherein coated layer is rumen-undegraded fat, is coated with the preparation method of slug particle
Including：
（1）Lysine salt and filler and optional sodium glutamate are well mixed, mixture is formed；
（2）To step（1）The optionally binder solution containing zytase is added in the mixture of acquisition, is well mixed,
Form softwood；
（3）By step（2）The softwood sieving of acquisition, forms coarse granule；
（4）By step（3）The coarse granule ball blast of acquisition, forms circular micropill；With
（5）Drying steps（4）The circular micropill of acquisition is simultaneously sieved, and forms coating slug particle.
Preferably wherein, step（1）In filler be microcrystalline cellulose and/or dextrin.
Preferably wherein, step（2）In adhesive be carboxymethyl cellulose.
Preferably wherein, step（2）In xylanase amino acid sequence such as SEQ ID NO：Shown in 2.
Preferably wherein, the weight ratio of lysine salt, filler, sodium glutamate and adhesive is 40 ~ 55：4~6：0~2：0.3~
0.8, preferably 42 ~ 50：4.5~5.5：0~1.5（Such as 0.5 ~ 1.5）：0.4 ~ 0.5, more preferably 43 ~ 48：4.8~5.2：0~1.2
（Such as 0.8 ~ 1.2）：0.45 ~ 0.55, most preferably 45 ~ 46：5：0~1：0.5.
Preferably wherein, coating slug particle and the weight ratio of coated layer are 40 ~ 60：40 ~ 60, preferably 46 ~ 56：43 ~ 53, more
Preferably 48.5 ~ 51.5：48.5 ~ 51.5, most preferably 1：1.
Preferably wherein, xylanase activity is 0 ~ 30IU, preferably 0 ~ 15IU in every gram of feed addictive（Such as 5 ~ 15IU）,
More preferably 0 ~ 10IU（Such as 8 ~ 10IU）, most preferably 0 ~ 9IU.
Preferably wherein, step（2）In sieve be 30 mesh sieves.
Preferably wherein, step（5）In screening be separation obtain can by 30 mesh sieves and by 40 mesh sieves retain particle.
The second of the present invention（2）Aspect, the invention provides the second of the present invention（1）The system of the feed addictive of aspect
Preparation Method, it includes the step being sprayed on rumen-undegraded fat on coating slug particle.
It is preferred that it includes：
（1）Lysine salt and filler and optional sodium glutamate are well mixed, mixture is formed；
（2）To step（1）The optionally binder solution containing zytase is added in the mixture of acquisition, is well mixed,
Form softwood；
（3）By step（2）The softwood sieving of acquisition, forms coarse granule；
（4）By step（3）The coarse granule ball blast of acquisition, forms circular micropill；
（5）Drying steps（4）The circular micropill of acquisition is simultaneously sieved, and forms coating slug particle；With
（6）Rumen-undegraded fat is sprayed on step（5）On the coating slug particle of acquisition.
The second of the present invention（3）Aspect, the invention provides the product containing 1B, it include coating slug particle and
The coated layer on coating slug particle is wrapped in, wherein coated layer is rumen-undegraded fat, coating slug particle includes lysine salt, filling
Agent and adhesive and optional sodium glutamate and optional zytase.
Preferably wherein, product is feed addictive.
Preferably wherein, lysine salt is lysine hydrochloride, lysine sulphate or their mixture, is preferably relied
Propylhomoserin sulfate.
Preferably wherein, filler is microcrystalline cellulose and/or dextrin.
Preferably wherein, adhesive is carboxymethyl cellulose.
Preferably wherein, xylanase amino acid sequence such as SEQ ID NO：Shown in 2.
Preferably wherein, the weight ratio of lysine salt, filler, sodium glutamate and adhesive is 40 ~ 55：4~6：0~2：0.3~
0.8, preferably 42 ~ 50：4.5~5.5：0~1.5（Such as 0.5 ~ 1.5）：0.4 ~ 0.5, more preferably 43 ~ 48：4.8~5.2：0~1.2
（Such as 0.8 ~ 1.2）：0.45 ~ 0.55, most preferably 45 ~ 46：5：0~1：0.5.
Preferably wherein, coating slug particle and the weight ratio of coated layer are 40 ~ 60：40 ~ 60, preferably 46 ~ 56：43 ~ 53, more
Preferably 48.5 ~ 51.5：48.5 ~ 51.5, most preferably 1：1.
Preferably wherein, often xylanase activity is 0 ~ 30IU, preferably 0 ~ 15IU in restraint product（Such as 5 ~ 15IU）, more preferably
For 0 ~ 10IU（Such as 8 ~ 10IU）, most preferably 0 ~ 9IU.
Preferably wherein, coating slug particle is the particle that can be retained by 30 mesh sieves and by 40 mesh sieves.
Preferably wherein, fat is preferably palm fat and/or lecithin, the production that can be more preferably obtained by commercial sources
Product, such as hundred Canons, Pepsi U.S., preferably hundred Canon HTL316, Pepsi U.S. T300 etc..In the embodiment of the present invention
In, coating fat used is Pepsi U.S. T300.
Such as the instruction without opposite or contradiction, aspects above can preferably present aspect or other each side herein
Content.
Three, the technique for preparing the product containing 1B
The 3rd of the present invention the（1）Aspect, the invention provides the preparation method of the product containing 1B, it includes：
（1）Lysine salt and filler and optional sodium glutamate are well mixed, mixture is formed；
（2）To step（1）The optionally binder solution containing zytase is added in the mixture of acquisition, is well mixed,
Form softwood；
（3）By step（2）The softwood sieving of acquisition（Such as, 30 mesh sieve）, form coarse granule；
（4）By step（3）The coarse granule ball blast of acquisition, forms circular micropill；
（5）Drying steps（4）The circular micropill of acquisition is simultaneously sieved, and forms coating slug particle grain；With,
（6）Rumen-undegraded fat is sprayed on step（5）On the coating slug particle of acquisition.
It is preferred that the product containing 1B prepared by the preparation method is the second of the present invention（3）Product described in aspect.
It is preferred that the preparation method is made up of following steps：
（1）Lysine salt and filler and optional sodium glutamate are well mixed, mixture is formed；
（2）To step（1）The optionally binder solution containing zytase is added in the mixture of acquisition, is well mixed,
Form softwood；
（3）By step（2）The softwood sieving of acquisition（Such as, 30 mesh sieve）, form coarse granule；
（4）By step（3）The coarse granule ball blast of acquisition, forms circular micropill；
（5）Drying steps（4）The circular micropill of acquisition is simultaneously sieved, and forms coating slug particle；With,
（6）Rumen-undegraded fat is sprayed on step（5）On the coating slug particle of acquisition.
The inventors discovered that, the step of to coarse granule ball blast to form circular micropill and/or dry circular micropill and carry out
Screening, can effectively improve the anti-degradation capability of the coated product containing 1B of rumen-undegraded fat.So ensuing
Aspect, the invention provides the 3rd of the present invention（1）Step in aspect（4）And/or step（5）Application.Throw preferably wherein
Ball is carried out in shot-blasting machine；And/or, screening is separated with the screen cloth of one or more special pore size distributions.
The 3rd of the present invention the（2）Aspect, the step of the invention provides to coarse granule ball blast to form circular micropill
Prepare the application in the method for the coated product containing 1B of rumen-undegraded fat.
The 3rd of the present invention the（3）Aspect, the invention provides dry circular micropill and sieved preparing rumen bypass
Application in the method for the coated product containing 1B of fat.
The 3rd of the present invention the（4）Aspect, the step of the invention provides to coarse granule ball blast to form circular micropill
Improve the application in the anti-degradation capability of the coated product containing 1B of rumen-undegraded fat.
The 3rd of the present invention the（5）Aspect, is being improved the invention provides dry circular micropill and the step of screening
Application in the anti-degradation capability of the coated product containing 1B of rumen-undegraded fat.
Preferably wherein, the step of to coarse granule ball blast to form circular micropill, is in（It is preferred that being next to）Formed coarse granule it
Afterwards.
Preferably wherein, dry circular micropill and be in the step of screening（It is preferred that being next to）Rumen-undegraded fat is sprayed on
Before being coated with slug particle.
Preferably wherein, the coated product containing 1B of rumen-undegraded fat is the second of the present invention（3）Described in aspect
Product.
Preferably wherein, the method for preparing the coated product containing 1B of rumen-undegraded fat is the 3 of the present invention（1）
Method described in aspect.
Preferably wherein, screening is that separation obtains the particle that can be retained by 30 mesh sieves and by 40 mesh sieves.
Such as the instruction without opposite or contradiction, aspects above can preferably present aspect or other each side herein
Content.
Four, application of the sodium glutamate in the product containing 1B
The inventors discovered that, a small amount of sodium glutamate is beneficial to the anti-degradation capability of the product containing 1B, and
Lysine is not influenceed to play the effect of feed.So, present invention also offers sodium glutamate in the product containing 1B
Using and correlation method.
The 4th of the present invention the（1）Aspect, the invention provides the preparation method of the product containing 1B, it includes：
（1）Lysine salt and filler and sodium glutamate are well mixed, mixture is formed；
（2）To step（1）The optionally binder solution containing zytase is added in the mixture of acquisition, is well mixed,
Form softwood；
（3）By step（2）The softwood sieving of acquisition（Such as, 30 mesh sieve）, form coarse granule；
（4）By step（3）The coarse granule ball blast of acquisition, forms circular micropill；
（5）Drying steps（4）The circular micropill of acquisition is simultaneously sieved, and forms coating slug particle；With,
（6）Rumen-undegraded fat is sprayed on step（5）On the coating slug particle of acquisition.
Preferably wherein, product is feed addictive.
Preferably wherein, step（1）In filler be microcrystalline cellulose and/or dextrin.
Preferably wherein, step（2）In adhesive be carboxymethyl cellulose.
Preferably wherein, step（2）In xylanase amino acid sequence such as SEQ ID NO：Shown in 2.
Preferably wherein, the weight ratio of lysine salt, filler, sodium glutamate and adhesive is 42 ~ 50：4.5~5.5：0.5~
1.5：0.4 ~ 0.5, more preferably 43 ~ 48：4.8~5.2：0.8~1.2：0.45 ~ 0.55, most preferably 45 ~ 46：5：1：0.5.
Preferably wherein, coating slug particle and the weight ratio of coated layer are 40 ~ 60：40 ~ 60, preferably 46 ~ 56：43 ~ 53, more
Preferably 48.5 ~ 51.5：48.5 ~ 51.5, most preferably 1：1.
Preferably wherein, xylanase activity is 0 ~ 30IU, preferably 0 ~ 15IU in every gram of feed addictive（Such as 5 ~ 15IU）,
More preferably 0 ~ 10IU（Such as 8 ~ 10IU）, most preferably 0 ~ 9IU.
Preferably wherein, xylanase activity is 0 ~ 30IU, preferably 0 ~ 15IU in every gram of feed addictive（Such as 5 ~ 15IU）,
More preferably 0 ~ 10IU（Such as 8 ~ 10IU）, most preferably 0 ~ 9IU.
Preferably wherein, step（5）In screening be separation obtain can by 30 mesh sieves and by 40 mesh sieves retain particle.
The 4th of the present invention the（2）Aspect, the invention provides the 4th of the present invention（1）Prepared by the method described in aspect
Product.
The 4th of the present invention the（3）Aspect, the invention provides sodium glutamate prepare rumen-undegraded fat it is coated contain L-
In the method for the product of lysine or in the anti-degradation capability for improving the coated product containing 1B of rumen-undegraded fat
Application.That is, the invention provides sodium glutamate in the method for preparing the coated product containing 1B of rumen-undegraded fat
Application, present invention also offers sodium glutamate improve the coated product containing 1B of rumen-undegraded fat anti-degraded energy
Application in power.
Preferably wherein, sodium glutamate is seldom, and the weight of such as sodium glutamate accounts for that rumen-undegraded fat is coated to be relied containing L-
The 0.5 ~ 1.5% of the product of propylhomoserin, preferably 0.8 ~ 1.2%, most preferably 1%.
Preferably wherein, the coated product containing 1B of rumen-undegraded fat is the 4th of the present invention（2）Described in aspect
Product.
Preferably wherein, the method for preparing the coated product containing 1B of rumen-undegraded fat is the 4 of the present invention（1）
Method described in aspect.
Such as the instruction without opposite or contradiction, aspects above can preferably present aspect or other each side herein
Content.
Five, the zytase of mutation and its application
The present inventor's surprisingly xylanase mutant, it is keeping the base of original high-temperature stability substantially
Enzymatic activity is added significantly on plinth, is particularly useful for being added into the feed addictive containing 1B.
The 5th of the present invention the（1）Aspect, the invention provides the zytase of mutation, it includes mutation Arg45Ser,
I.e. the 45th amino acids residue sports Ser by Arg.
It is preferred that the amino acid sequence of the zytase of the mutation
（1）Such as SEQ ID No:Shown in 2；Or
（2）It is to SEQ ID No:2 lack, add and/or replace one or several（Preferably more than 7, more preferably not
More than 5,3 are such as no more than）Amino acid residue and the reservation SEQ ID No obtained:The sequence of 2 activity, and wherein include
It is mutated Arg45Ser.
The 5th of the present invention the（2）Aspect, the invention provides the 5th of the coding present invention（1）Xylan described in aspect
The gene of enzyme.
It is preferred that the nucleotide sequence of the gene such as SEQ ID No:Shown in 1.
The 5th of the present invention the（3）Aspect, the invention provides include the 5th of the present invention（2）Gene described in aspect
Carrier.
The 5th of the present invention the（4）Aspect, the invention provides strain, it includes the 5th of the present invention（2）Described in aspect
Gene, or with the present invention the 5th（3）Carrier described in aspect converts or transfects and obtain.
It is preferred that the strain is secreting, expressing bacterium, and e.g., yeast.
The 5th of the present invention the（5）Aspect, the invention provides the 5th of the fermenting and producing present invention（1）Wood described in aspect
The method of dextranase, it includes cultivating the 5th of the present invention under fermentation conditions（4）Strain described in aspect, divides from culture
Separate out the 5th of the present invention（1）Zytase described in aspect.
The 5th of the present invention the（6）Aspect, the invention provides the 5th of the present invention（1）Zytase described in aspect exists
Prepare the purposes in the product containing 1B.
Preferably wherein, the of the invention 5th in every gram of product containing 1B（1）Xylanase activity described in aspect
For 5 ~ 15IU, most preferably more preferably 8 ~ 10IU, 9IU.
Preferably wherein, product is feed addictive.
Preferably wherein, the coated layer that the product containing 1B includes coating slug particle and is wrapped on coating slug particle,
Wherein coated layer is rumen-undegraded fat, and coating slug particle includes the 5th of the present invention（1）Zytase, lysine described in aspect
Salt, filler and adhesive and optional sodium glutamate.
More preferably wherein, lysine salt is lysine hydrochloride, lysine sulphate or their mixture, is preferably
Lysine sulphate.
More preferably wherein, filler is microcrystalline cellulose and/or dextrin.
More preferably wherein, adhesive is carboxymethyl cellulose.
More preferably wherein, xylanase amino acid sequence such as SEQ ID NO：Shown in 2.
Preferably wherein, the weight ratio of lysine salt, filler, sodium glutamate and adhesive is 40 ~ 55：4~6：0~2：0.3~
0.8, preferably 42 ~ 50：4.5~5.5：0~1.5（Such as 0.5 ~ 1.5）：0.4 ~ 0.5, more preferably 43 ~ 48：4.8~5.2：0~1.2
（Such as 0.8 ~ 1.2）：0.45 ~ 0.55, most preferably 45 ~ 46：5：0~1：0.5.
Preferably wherein, coating slug particle and the weight ratio of coated layer are 40 ~ 60：40 ~ 60, preferably 46 ~ 56：43 ~ 53, more
Preferably 48.5 ~ 51.5：48.5 ~ 51.5, most preferably 1：1.
Preferably wherein, coating slug particle is the particle that can be retained by 30 mesh sieves and by 40 mesh sieves.
Preferably wherein, fat is preferably palm fat and/or lecithin, the production that can be more preferably obtained by commercial sources
Product, such as hundred Canons, Pepsi U.S., preferably hundred Canon HTL316, Pepsi U.S. T300 etc..In the embodiment of the present invention
In, coating fat used is Pepsi U.S. T300.
Preferably wherein, the method for preparation includes：
（1）Lysine salt and filler and optional sodium glutamate are well mixed, mixture is formed；
（2）To step（1）The binder solution containing zytase is added in the mixture of acquisition, is well mixed, is formed
Softwood；
（3）By step（2）The softwood sieving of acquisition（Such as, 30 mesh sieve）, form coarse granule；
（4）By step（3）The coarse granule ball blast of acquisition, forms circular micropill；
（5）Drying steps（4）The circular micropill of acquisition is simultaneously sieved, and forms coating slug particle；With,
（6）Optionally, rumen-undegraded fat is sprayed on step（5）On the coating slug particle of acquisition.
Such as the instruction without opposite or contradiction, aspects above can preferably present aspect or other each side herein
Content.
The invention has the advantages that：By the improvement of many aspects, product safety of the invention is reliable, relative to
Prior art, can more effectively reduce the rate of release of lysine in animal body, and anti-degradation capability is stronger, more improve the profit of feed
Promote growing for animal with rate.
In order to make it easy to understand, the present invention will be described in detail by specific embodiment below.Need to refer in particular to
Go out, the description that these descriptions are merely exemplary, and be not meant to limit the scope of the invention.Opinion according to this specification
State, many changes of the invention, change and will be apparent from for one of ordinary skill in the art.
In addition, the present invention refer to open source literature, these documents are their full text in order to more clearly describe the present invention
Content is included and referred to herein, just looks like that their full text is repeated and was expressly recited the same herein.
Embodiment
Present disclosure is further illustrated by the following examples.As do not specialized, technology used in embodiment
Conventional meanses that means are well known to those skilled in the art and commercially available common instrument, reagent, reference can be made to《Molecular Cloning: A Laboratory
Guide（3rd edition）》（Science Press）、《Microbiology Experiment（4th edition）》（Higher Education Publishing House）And corresponding instrument and
Manufacturers instruction of reagent etc. is referred to.
The preparation of the expression construct of the xylanase gene of the saltant type of embodiment 1
According to us in the xylanase gene of preceding No. 201210185883 records of Chinese Patent No., the 45th will be encoded thereon
The Arg of position is mutated into Ser, i.e., the CGC of nucleotide sequence 133-135 is mutated into TCC, the core of the zytase of saltant type
The SEQ ID No of nucleotide sequence such as sequence table:Shown in 1, the amino acid sequence such as SEQ ID No of coded zytase:2 institutes
Show.Then, according to《Molecular Cloning:A Laboratory guide》And the operating guidance of commercial reagents used builds the yeast of secreting, expressing,
In brief, the gene SEQ ID No of forward primer such as sequence table of the zytase will be encoded:Shown in 3（Introduce
EcoR I restriction enzyme sites）, reverse primer such as sequence table SEQ ID No:Shown in 4（Introduce Not I restriction enzyme sites）Expand
After increasing use EcoR I and Not I double digestions, and with the pPIC3.5 plasmids through the two endonuclease digestions（It is purchased from
Invitrogen companies）It is attached, is transformed into bacillus coli DH 5 alpha with T4 DNA ligases.Choose positive colony, extract
Go out plasmid therein, through sequence verification it is correct after, by positive plasmid with after Sal I linearization for enzyme restriction, converted by electrotransformation
Enter GS115 plants of pichia yeast, positive yeast transformant is chosen on the flat board containing G418, is returned.
The fermentation preparation and determination of activity of the zytase of the saltant type of embodiment 2
The positive yeast transformant that embodiment 1 is obtained is inoculated into 50ml BMGY fluid nutrient mediums（100mM potassium phosphates delay
Fliud flushing（pH6.0）, wherein containing 1% yeast extract, 2% peptone, Yeast Nitrogen base 1.34%, biotin 4*10-5%, glucose
2%, glycerine 1%）In, cultivated in 30 DEG C, 200rpm to OD600 and reach 5.0.
Above-mentioned culture is transferred in 1L BMGY fluid nutrient mediums, is cultivated 24 hours in 30 DEG C, 200rpm, adds 5ml
Methanol, then proceedes to cultivate 8 days in 30 DEG C, 200rpm, adds within during which every 24 hours 5ml methanol, ventilation control dissolved oxygen（DO）Greatly
In 20%, and it is 6.0 to control pH（Adjusted with ammoniacal liquor）.After the completion of culture, with 0.22 μm of membrane filtration and retain supernatant, on this
Clear liquid is detected through SDS-PAGE is wherein rich in the corresponding protein of zytase, and thus fermentation obtains zytase supernatant.
Fresh supernatant is taken, is divided into three parts, portion refrigeration is stand-by（A）, potassium phosphate is used after portion freeze-drying to solid
Buffer solution（pH6.0）Again dissolve（B）, it is a to use kaliumphosphate buffer after evaporation drying to solid in 80 DEG C of baking ovens
（pH6.0）Again dissolve（C）；Control uses phosphoric acid using the xylanase gene of No. 201210185883 discoveries of Chinese Patent No.
Potassium buffer solution（pH6.0）It is divided into three parts after dissolving, portion refrigeration is stand-by（CA）, another two parts are freeze-dried respectively（CB）With 80 DEG C of bakings
Case evaporation drying（CC）, kaliumphosphate buffer is then dissolved in again（pH6.0）.These xylanase activities are determined respectively, are converted into
The enzyme activity of original unit's volume supernatant or Unit Weight, is calculated due to the enzyme activity loss late that high temperature dehydration is caused, as a result such as the institute of table 1
Show, show relative to it is original we have found that high temperature resistant dehydration zytase, the zytase of saltant type of the invention, although
High temperature resistant water separation capability is slightly worse than original, but still remains more than 75% activity, especially surprisingly, enzyme activity
Property, which has, to be obviously improved, even if by high temperature dehydration, the absolute value of its enzymatic activity is still above original zytase, even
Higher than enzyme activity of original zytase when without high temperature dehydration.
The high temperature dehydration enzyme activity loss late of the zytase of the invention and commercially available of table 1
| Sample | Enzyme activity | Loss late | Sample | Enzyme activity | Loss late |
| A | 3.7*104IU/L | CA | 2.4*104IU/L | ||
| B | 3.5*104IU/L | 5.4% | CB | 2.2*104IU/L | 8.3% |
| C | 2.8*104IU/L | 24% | CC | 2.0*104IU/L | 17% |
The coating of the 1B of embodiment 3
The method recorded according to us in preceding Chinese patent 201210440083, it is 63.5% to obtain 1B content（w/
w）The sulfate containing 1B granular core（Itself it is commercially pure 1B sulfate）, no longer with low-purity
1B outer layer covers, directly as the lysine raw material of the feed of the present invention.
The formula of the feed addictive of the present invention of table 2
| Formula 1 | Formula 2 | Formula 3 | Formula 4 | |
| Lysine raw material | 46kg | 45kg | 46kg | 45kg |
| Sodium glutamate | / | 1kg | / | 1kg |
| Microcrystalline cellulose | 4kg | 4kg | 4kg | 4kg |
| Dextrin | 1kg | 1kg | 1kg | 1kg |
| Carboxymethyl cellulose（CMC） | 0.5 | 0.5 | 0.5 | 0.5 |
| Zytase | / | / | 9*105IU | 9*105IU |
| Rumen-undegraded fat powder | 48.5kg | 48.5kg | 48.5kg | 48.5kg |
As shown in table 2, detailed process is as follows for coating raw material used：
By lysine raw material, microcrystalline cellulose and the dextrin of formula ratio and（When formula contains sodium glutamate）Paddy
Propylhomoserin sodium pours into trough type mixing machine, is well mixed, and obtains coating core mixture；
By the carboxymethyl cellulose of formula ratio 25L water（When formula does not contain zytase）Or according to implementation
Zytase supernatant prepared by the method for example 2（When formula contains zytase）Soak, be configured to 2% CMC solution,
Then directly pour into above-mentioned coating core mixture, be well mixed, form softwood；
Above-mentioned softwood is poured into swing comminutor, through 30 eye mesh screens（Aperture is 0.6mm）, diameter, which is made, is about
0.6mm coarse granule；
Above-mentioned coarse granule is poured into ball blast 3 minutes in the ball-type shot-blasting machine run with 2200rpm, circular micropill is formed；
Above-mentioned micropill is placed in the fluidized granulation seed-coating machine of 60 DEG C of EAT and dried 45 minutes, then respectively through 30
Eye mesh screen（Aperture is 0.6mm）With 40 eye mesh screens（Aperture is 0.425mm）, reservation can be by 30 eye mesh screens and by 40 mu of screen clothes section
The particle diameter stayed is 0.425-0.6mm coating slug particle；
The rumen-undegraded fat powder of formula ratio is placed in oil tank and is heated to 90 DEG C, until completely melted, in 80 DEG C of insulations simultaneously
The shower nozzle for connecting fluidized granulation seed-coating machine is sprayed into fluidized granulation seed-coating machine, is sprayed on above-mentioned coating slug particle surface therein, its
Middle fluidized granulation seed-coating machine air-introduced machine rotating speed 3500rpm, 60 DEG C of EAT finally respectively obtains coating slug particle and rumen bypass
Fatty powder weight ratio about 1：1 above-mentioned four kinds of formulas distinguish corresponding coated 1B feed addictive.
In addition, being prepared according to the formula of formula 1 substantially according to said process, different is to omit ball blast step, i.e.,
Directly coarse granule is placed in the fluidized granulation seed-coating machine of 60 DEG C of EAT and dried, the 1B for obtaining control technique 1 is raised
Feed additives；
Prepared according to the formula of formula 1 substantially according to said process, different is that two steps are sieved after omission is dried
Step, i.e., after being dried 45 minutes in the fluidized granulation seed-coating machine of 60 DEG C of EAT, do not sieve, directly by all particles all
Proceed next step coating, obtain the 1B feed addictive of control technique 2.
The degraded test of the coated 1B feed addictive of embodiment 4
According to Zhao He etc.（Different proportion sheep's hay and the research China of ensilage dry matter degradability in cow rumen
Ox industry science, 34 (2):13-16）Description internal Nylon Bag, in adult dairy cattle to according to the method for embodiment 3 prepare it is each
Plant coated feed addictive and carry out degraded test, testing index is used as using wherein 1B.In the case of not coated,
The degradation rate of 1B is up to more than 95% after 2 hours（That is 1B conservation rate is less than 5%）, and coated feed addictive
Conservation rate is as shown in table 3 below：
The anti-degradation effect of the feed addictive of the present invention of table 3
As shown in table 3, but if the method recorded relative to us in preceding Chinese patent 201210440083, the present invention
Coating technique can further extend remaining time of the 1B in animal stomach, wherein whether with the addition of the present invention wood
Dextranase influences little for encapsulating effect, although it was unexpectedly determined that ball blast and the technique of screening are very small, but adopt
Conservation rate about 10% can be improved after, more surprisingly, if wherein adding a small amount of sodium glutamate（If only added
Plus glutamic acid, effect will significantly be not so good as its sodium salt）, will make it that coated particle is not allowed more easily rupturable so that bad ammonia therein
Acid release is more smooth, either in short term（In 2 hours）Or it is long-term（Such as 24 or 48 hours）, all significantly improve lysine
Anti- degradation effect in cud.
Effectiveness of the coated 1B feed addictive of embodiment 5 to calf growth effect
Commission herding research institute of Ningxia academy of agricultural sciences is fed with March with all feeds additive prepared according to the method for embodiment 3
The calf in age 8 weeks, to be fed with the feed addictive without the present invention（Only feed basal diet）With with the addition of commercially available non-packet
The equivalent lysine additives for forage of quilt is control.As a result it is as shown in table 4 below：
Growth effect of the feed addictive of the present invention of table 4 to calf
| Feed | Initial average weight（kg） | Terminate average weight（kg） | Increased weight（kg） | Increased weight increase rate（%） |
| Basal diet | 125.7 | 165.3 | 39.6 | / |
| Basal diet+not coated lysine | 126.1 | 166.9 | 40.8 | 3.0 |
| Basal diet+formula 1 | 125.6 | 169.5 | 43.9 | 10.9 |
| Basal diet+formula 2 | 126.4 | 170.5 | 44.1 | 11.4 |
| Basal diet+formula 3 | 125.8 | 171.0 | 45.2 | 14.1 |
| Basal diet+formula 4 | 124.8 | 170.3 | 45.5 | 14.9 |
As shown in table 4, growth of the feeding 1B to calf has certain facilitation, but if directly feeds
Raise, its facilitation is very limited, only less than 5%, and the feed of equivalent lysine is added after being coated with by the method for the present invention
Additive, can remarkably promote growth, improve more than 10%, more can be further after the zytase that especially with the addition of the present invention
Promote growth about 4%.Due to being fed with frequency in experiment（Twice daily）It is higher, therefore the longer time of some formulas of the present invention
Encapsulating conservation rate advantage embody not to be in the influence for growing calf very fully, but still old 0.5% and more than
Lifting, it is contemplated that it is actual raise in can embody more notable.In addition, during experiment, without adverse reaction report, showing the present invention
Formula be safe.
<110>Ningbo Eppen Biotech Co., Ltd.
<120>The zytase of mutation and its application in 1B product
<130>Electronic application
<160> 5
<170> PatentIn version 3.3
<210> 1
<211> 1428
<212> DNA
<213>Artificial sequence
<400> 1
atgattagcc tggcggcgag cctgccgatt ccgctggtga gcgcgctgcc gcatgcgcat 60
attagcgtga acgcgcatac ccgctttcag caggtggatg gctttggctt tagcgaagcg 120
tttcagcgcg cgtccgatat ttatggcaaa gatggcctga gcccggaaga tcgcacccgc 180
gtgctggatc tgctgtttag cgatacccgc ggcgcgggcc tgaccattgt gcgcaacggc 240
attggcagca acagcaccat taaagatttt atgaacagca ttgaaccgtt tagcccgggc 300
agcccgagcg cgccgccgca ttatgtgtgg gatcgcaacg atagcggcca ggtgtggctg 360
agccatgaag cggcgagcta tggcgtgaac accttttatg cggatgcgtg gagcgcgccg 420
ggctatatga aaaccaacga tgatgatagc aacggcggct atctgtgcgg cgtgagcaac 480
accagctgcg cgagcggcga ttggaaacag gcgtatgcga actatctggt gcagtatatt 540
catttttatc agcaggtggg cattaaagtg acccatctgg gctttctgaa cgaaccgcag 600
gaagatgtga cctatgcgag catgctgagc gatggcaccc aggcggcggc ggattttatt 660
aaagtgctgg cgccgaccgt gaaagcggcg ggcctggatg tgaaactgac ctgctgcgat 720
ggcgtgggct gggaagaaca gcgcgcgatg ctgccgggcc tgcaggcggg cggcccggaa 780
catagcgcgg aaagctatct gagcgtgatt accgcgcatg gctataacag cccgccgacc 840
accccgctgg aaaccagcct gccggtgtgg atgaccgaat gggcggatct gaacgataac 900
tataccgcgg cgtggtatga aaacggcgcg ccgggcgaag gcctgacctg ggcgaaccgc 960
attcaggatg cgtttacccg cagcaacgtg agcgcgtttc tgcattggat tggcgcggaa 1020
aacggcacca gcaacagccc gctgattaac ctgaacggcg atagctatgt ggcgagcaaa 1080
cgcctgtggg cgtttggcca gtttagccgc tttgtgcgcc cgggcgcggt gcgcattgat 1140
gcggcgagca gcgatccgct ggtgaccgtg agcgcgtttc agaacaaaga aaacggcgtg 1200
gtggcgaccc aggtgattaa caacgcggat accgattatg aagtggaagt ggatctgacc 1260
ggccgcggcc atctggtgag cgtggtgcag ccgtatctga ccaacaacga acatgatctg 1320
gaagcgagca gcccgattat taccctggcg ctgggcgcgg cgaccaaatt taccagcatt 1380
gtgccggcgc gcagcctggt gagctttgtg agcaaaaccg cgctgtaa 1428
<210> 2
<211> 475
<212> PRT
<213>Artificial sequence
<400> 2
Met Ile Ser Leu Ala Ala Ser Leu Pro Ile Pro Leu Val Ser Ala Leu
1 5 10 15
Pro His Ala His Ile Ser Val Asn Ala His Thr Arg Phe Gln Gln Val
20 25 30
Asp Gly Phe Gly Phe Ser Glu Ala Phe Gln Arg Ala Ser Asp Ile Tyr
35 40 45
Gly Lys Asp Gly Leu Ser Pro Glu Asp Arg Thr Arg Val Leu Asp Leu
50 55 60
Leu Phe Ser Asp Thr Arg Gly Ala Gly Leu Thr Ile Val Arg Asn Gly
65 70 75 80
Ile Gly Ser Asn Ser Thr Ile Lys Asp Phe Met Asn Ser Ile Glu Pro
85 90 95
Phe Ser Pro Gly Ser Pro Ser Ala Pro Pro His Tyr Val Trp Asp Arg
100 105 110
Asn Asp Ser Gly Gln Val Trp Leu Ser His Glu Ala Ala Ser Tyr Gly
115 120 125
Val Asn Thr Phe Tyr Ala Asp Ala Trp Ser Ala Pro Gly Tyr Met Lys
130 135 140
Thr Asn Asp Asp Asp Ser Asn Gly Gly Tyr Leu Cys Gly Val Ser Asn
145 150 155 160
Thr Ser Cys Ala Ser Gly Asp Trp Lys Gln Ala Tyr Ala Asn Tyr Leu
165 170 175
Val Gln Tyr Ile His Phe Tyr Gln Gln Val Gly Ile Lys Val Thr His
180 185 190
Leu Gly Phe Leu Asn Glu Pro Gln Glu Asp Val Thr Tyr Ala Ser Met
195 200 205
Leu Ser Asp Gly Thr Gln Ala Ala Ala Asp Phe Ile Lys Val Leu Ala
210 215 220
Pro Thr Val Lys Ala Ala Gly Leu Asp Val Lys Leu Thr Cys Cys Asp
225 230 235 240
Gly Val Gly Trp Glu Glu Gln Arg Ala Met Leu Pro Gly Leu Gln Ala
245 250 255
Gly Gly Pro Glu His Ser Ala Glu Ser Tyr Leu Ser Val Ile Thr Ala
260 265 270
His Gly Tyr Asn Ser Pro Pro Thr Thr Pro Leu Glu Thr Ser Leu Pro
275 280 285
Val Trp Met Thr Glu Trp Ala Asp Leu Asn Asp Asn Tyr Thr Ala Ala
290 295 300
Trp Tyr Glu Asn Gly Ala Pro Gly Glu Gly Leu Thr Trp Ala Asn Arg
305 310 315 320
Ile Gln Asp Ala Phe Thr Arg Ser Asn Val Ser Ala Phe Leu His Trp
325 330 335
Ile Gly Ala Glu Asn Gly Thr Ser Asn Ser Pro Leu Ile Asn Leu Asn
340 345 350
Gly Asp Ser Tyr Val Ala Ser Lys Arg Leu Trp Ala Phe Gly Gln Phe
355 360 365
Ser Arg Phe Val Arg Pro Gly Ala Val Arg Ile Asp Ala Ala Ser Ser
370 375 380
Asp Pro Leu Val Thr Val Ser Ala Phe Gln Asn Lys Glu Asn Gly Val
385 390 395 400
Val Ala Thr Gln Val Ile Asn Asn Ala Asp Thr Asp Tyr Glu Val Glu
405 410 415
Val Asp Leu Thr Gly Arg Gly His Leu Val Ser Val Val Gln Pro Tyr
420 425 430
Leu Thr Asn Asn Glu His Asp Leu Glu Ala Ser Ser Pro Ile Ile Thr
435 440 445
Leu Ala Leu Gly Ala Ala Thr Lys Phe Thr Ser Ile Val Pro Ala Arg
450 455 460
Ser Leu Val Ser Phe Val Ser Lys Thr Ala Leu
465 470 475
<210> 3
<211> 22
<212> DNA
<213> Artificial
<220>
<223>Forward primer
<400> 3
cgaattcgat gattagcctg gc 22
<210> 4
<211> 25
<212> DNA
<213> Artificial
<220>
<223>Reverse primer
<400> 4
agcggccgca ttacagcgcg gtttt 25
Claims (16)
1. the zytase of mutation, its amino acid sequence such as SEQ ID No:Shown in 2.
2. encode the gene of the zytase described in claim 1.
3. the gene described in claim 2, its nucleotide sequence such as SEQ ID No:Shown in 1.
4. include the carrier of the gene described in Claims 2 or 3.
5. strain, it includes the gene described in Claims 2 or 3, or with the carrier conversion or transfection described in claim 4
.
6. the strain described in claim 5, it is secreting, expressing bacterium.
7. the strain described in claim 6, it is yeast.
8. the method for the zytase described in fermenting and producing claim 1, it includes culture claim 5-7 under fermentation conditions
One of described in strain, the zytase described in claim 1 is isolated from culture.
9. purposes of the zytase in the product containing 1B is prepared described in claim 1.
10. the purposes described in claim 9, wherein product are feed addictives.
11. the purposes described in claim 9, wherein the product containing 1B includes coating slug particle and is wrapped in coating core
Coated layer on grain, wherein coated layer are rumen-undegraded fats, coating slug particle include claim 1 described in zytase, rely
Propylhomoserin salt, filler and adhesive and sodium glutamate.
12. the purposes described in claim 9, wherein the zytase in every gram of product containing 1B described in claim 1
Activity is 5 ~ 15IU.
13. the purposes described in claim 12, wherein the zytase in every gram of product containing 1B described in claim 1
Activity is 8 ~ 10IU.
14. the purposes described in claim 13, wherein the zytase in every gram of product containing 1B described in claim 1
Activity is 9IU.
15. the purposes described in claim 9, wherein the method prepared includes：
（1）Lysine salt and filler and sodium glutamate are well mixed, mixture is formed；
（2）To step（1）The binder solution containing zytase is added in the mixture of acquisition, is well mixed, softwood is formed；
（3）By step（2）The softwood sieving of acquisition, forms coarse granule；
（4）By step（3）The coarse granule ball blast of acquisition, forms circular micropill；
（5）Drying steps（4）The circular micropill of acquisition is simultaneously sieved, and forms coating slug particle；With,
（6）Optionally, rumen-undegraded fat is sprayed on step（5）On the coating slug particle of acquisition.
16. the purposes described in claim 15, wherein step（3）In sieve be 30 mesh sieves.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410777733.6A CN104480089B (en) | 2014-12-17 | 2014-12-17 | The zytase of mutation and its application in L lysine products |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410777733.6A CN104480089B (en) | 2014-12-17 | 2014-12-17 | The zytase of mutation and its application in L lysine products |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104480089A CN104480089A (en) | 2015-04-01 |
| CN104480089B true CN104480089B (en) | 2017-07-18 |
Family
ID=52754697
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410777733.6A Active CN104480089B (en) | 2014-12-17 | 2014-12-17 | The zytase of mutation and its application in L lysine products |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104480089B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102763771A (en) * | 2012-06-07 | 2012-11-07 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and xylanase |
| CN103355502A (en) * | 2012-06-07 | 2013-10-23 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and compound enzyme |
| CN104039959A (en) * | 2011-11-11 | 2014-09-10 | 诺维信股份有限公司 | Polypeptides having xylanase activity and polynucleotides encoding same |
-
2014
- 2014-12-17 CN CN201410777733.6A patent/CN104480089B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104039959A (en) * | 2011-11-11 | 2014-09-10 | 诺维信股份有限公司 | Polypeptides having xylanase activity and polynucleotides encoding same |
| CN102763771A (en) * | 2012-06-07 | 2012-11-07 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and xylanase |
| CN103355502A (en) * | 2012-06-07 | 2013-10-23 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and compound enzyme |
| CN103444984A (en) * | 2012-06-07 | 2013-12-18 | 宁夏伊品生物科技股份有限公司 | Lysine and xylanase fermented and coated feed |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104480089A (en) | 2015-04-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103652452B (en) | A kind of prawn biological feedstuff and application thereof | |
| CN106260662A (en) | A kind of preparation method of enzymatic hydrolysis and fermentation type premix material | |
| CN101156659A (en) | Organic swine core premix | |
| CN107208080A (en) | Composition including the polypeptide with xylanase activity and the polypeptide with nofuranosidase activity | |
| CN101904432B (en) | Feed for enhancing disease resistance of piglets and accelerating growth of piglets and preparation method thereof | |
| CN103734501A (en) | Grass carp feed, and preparation method thereof | |
| CN103734549A (en) | Large-grain milk replacer and preparing method thereof | |
| CN105146173A (en) | Straw mixed particle fodder for sheep and preparing method | |
| CN104335910A (en) | Novel edible pig house pad materials and preparation method of pig house pad bed | |
| CN109153981A (en) | Polypeptide with alpha-galactosidase activity and the polynucleotides for encoding it | |
| CN109196098A (en) | Composition comprising the polypeptide with galactanase activity and the polypeptide with betagalactosidase activity | |
| CN104605155B (en) | A kind of Caragana Microphylla biological composite granular feedstuff | |
| CN106720973A (en) | A kind of organic standard pig premix and preparation method thereof | |
| CN104472877B (en) | The preparation technology of L lysine products | |
| CN106417912A (en) | Pleurotus fungus residue feed and preparation method thereof | |
| CN105861615A (en) | Production method of amino acid bioactive peptide for aquatic products | |
| CN109497262A (en) | Utilize the method for wet basis fermentation raw material preparation sheep feed and the sheep feed of preparation | |
| CN104431369B (en) | Rumen-protected lysine feed | |
| CN108740528A (en) | Kind geese in laying period feed and preparation method thereof and the method for breeding for planting goose | |
| CN104221885A (en) | Preparation method of novel hog house padding having food calling effect and bedding with novel hog house padding | |
| CN104397333A (en) | Production method of selenium-enriched ryegrass hay meal feed | |
| CN109312321A (en) | Composition comprising the polypeptide with galactanase activity and the polypeptide with betagalactosidase activity | |
| CN104480089B (en) | The zytase of mutation and its application in L lysine products | |
| CN109415709A (en) | Polypeptide with xylanase activity and the polynucleotides for encoding it | |
| CN106316533A (en) | Culture medium of lentinula edodes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Mutational xylanase and application thereof in L-lysine product Effective date of registration: 20190201 Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd.|INNER MONGOLIA EPPEN BIOTECH Co.,Ltd. Registration number: 2019640000001 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20200116 Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd. Registration number: 2019640000001 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Mutational xylanase and application thereof in L-lysine product Effective date of registration: 20200120 Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd. Registration number: Y2020640000002 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20210126 Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd. Registration number: Y2020640000002 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Mutant xylanase and its application in L-lysine products Effective date of registration: 20210127 Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd. Registration number: Y2021640000001 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20220129 Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd. Registration number: Y2021640000001 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Mutant xylanase and its application in L-lysine products Effective date of registration: 20220323 Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd. Registration number: Y2022640000002 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20230315 Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd.|INNER MONGOLIA EPPEN BIOTECH Co.,Ltd.|Heilongjiang Yipin Biotechnology Co.,Ltd. Registration number: Y2022640000002 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Mutated Xylanase and Its Application in L-Lysine Products Granted publication date: 20170718 Pledgee: Yongning County Branch of China Agricultural Development Bank Pledgor: NINGXIA EPPEN BIOTECH Co.,Ltd.|INNER MONGOLIA EPPEN BIOTECH Co.,Ltd.|Heilongjiang Yipin Biotechnology Co.,Ltd. Registration number: Y2024640000005 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |