CN109072270A - 由β葡聚糖、低聚糖或葡糖产生的新型β-1,3-1,6-内切葡聚糖酶 - Google Patents
由β葡聚糖、低聚糖或葡糖产生的新型β-1,3-1,6-内切葡聚糖酶 Download PDFInfo
- Publication number
- CN109072270A CN109072270A CN201780026365.3A CN201780026365A CN109072270A CN 109072270 A CN109072270 A CN 109072270A CN 201780026365 A CN201780026365 A CN 201780026365A CN 109072270 A CN109072270 A CN 109072270A
- Authority
- CN
- China
- Prior art keywords
- oligosaccharide
- glucose
- endoglucanase
- kelp
- generating
- 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.)
- Granted
Links
- 229920001542 oligosaccharide Polymers 0.000 title claims abstract description 81
- 150000002482 oligosaccharides Chemical class 0.000 title claims abstract description 81
- 108010059892 Cellulase Proteins 0.000 title claims abstract description 53
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 title claims abstract description 51
- 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 title claims abstract description 48
- 239000008103 glucose Substances 0.000 title claims abstract description 48
- 229920002498 Beta-glucan Polymers 0.000 title abstract description 11
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 title abstract description 10
- 241000512259 Ascophyllum nodosum Species 0.000 claims abstract description 54
- 238000006116 polymerization reaction Methods 0.000 claims abstract description 9
- 239000000758 substrate Substances 0.000 claims description 45
- DBTMGCOVALSLOR-DEVYUCJPSA-N (2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](CO)O[C@H](O)[C@@H]2O)O)O[C@H](CO)[C@H]1O DBTMGCOVALSLOR-DEVYUCJPSA-N 0.000 claims description 42
- 239000005717 Laminarin Substances 0.000 claims description 42
- 229920001543 Laminarin Polymers 0.000 claims description 42
- 239000000203 mixture Substances 0.000 claims description 16
- 108090000623 proteins and genes Proteins 0.000 claims description 16
- 230000001580 bacterial effect Effects 0.000 claims description 14
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 108010001682 Dextranase Proteins 0.000 claims description 12
- 241001670248 Saccharophagus degradans Species 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 11
- 241001071795 Gentiana Species 0.000 claims description 8
- 230000014509 gene expression Effects 0.000 claims description 8
- 239000013598 vector Substances 0.000 claims description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 2
- 239000002253 acid Substances 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 229910021529 ammonia Inorganic materials 0.000 claims 1
- 150000002016 disaccharides Chemical class 0.000 claims 1
- 210000000232 gallbladder Anatomy 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 37
- 230000013595 glycosylation Effects 0.000 abstract description 10
- 238000006206 glycosylation reaction Methods 0.000 abstract description 10
- 230000012666 negative regulation of transcription by glucose Effects 0.000 abstract 1
- 102000004190 Enzymes Human genes 0.000 description 27
- 108090000790 Enzymes Proteins 0.000 description 27
- 229940088598 enzyme Drugs 0.000 description 27
- 238000012790 confirmation Methods 0.000 description 14
- 210000004027 cell Anatomy 0.000 description 13
- 108020004414 DNA Proteins 0.000 description 10
- 150000001413 amino acids Chemical group 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 10
- 238000004128 high performance liquid chromatography Methods 0.000 description 9
- 150000007523 nucleic acids Chemical class 0.000 description 9
- 238000004809 thin layer chromatography Methods 0.000 description 9
- 101710130619 Glucan endo-1,3-beta-glucosidase Proteins 0.000 description 8
- 101710184061 Glucan endo-1,3-beta-glucosidase, acidic isoform Proteins 0.000 description 8
- 101710180749 Glucan endo-1,3-beta-glucosidase, basic isoform Proteins 0.000 description 8
- 230000002255 enzymatic effect Effects 0.000 description 8
- 238000006911 enzymatic reaction Methods 0.000 description 8
- 150000004676 glycans Chemical class 0.000 description 8
- 108010070643 prolylglutamic acid Proteins 0.000 description 8
- 229920001503 Glucan Polymers 0.000 description 7
- 241000588724 Escherichia coli Species 0.000 description 6
- 239000007795 chemical reaction product Substances 0.000 description 6
- 108020004707 nucleic acids Proteins 0.000 description 6
- 102000039446 nucleic acids Human genes 0.000 description 6
- 229920001282 polysaccharide Polymers 0.000 description 6
- 239000005017 polysaccharide Substances 0.000 description 6
- 238000000074 matrix-assisted laser desorption--ionisation tandem time-of-flight detection Methods 0.000 description 5
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 4
- QIGJYVCQYDKYDW-UHFFFAOYSA-N 3-O-alpha-D-mannopyranosyl-D-mannopyranose Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(CO)OC(O)C1O QIGJYVCQYDKYDW-UHFFFAOYSA-N 0.000 description 4
- DBTMGCOVALSLOR-UHFFFAOYSA-N 32-alpha-galactosyl-3-alpha-galactosyl-galactose Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(OC2C(C(CO)OC(O)C2O)O)OC(CO)C1O DBTMGCOVALSLOR-UHFFFAOYSA-N 0.000 description 4
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 4
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 4
- 229930045534 Me ester-Cyclohexaneundecanoic acid Natural products 0.000 description 4
- LGSANCBHSMDFDY-GARJFASQSA-N Pro-Glu-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCC(=O)O)C(=O)N2CCC[C@@H]2C(=O)O LGSANCBHSMDFDY-GARJFASQSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- DBTMGCOVALSLOR-AKJQSPAISA-N beta-D-Glcp-(1->3)-beta-D-Glcp-(1->3)-beta-D-Glcp Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](CO)O[C@@H](O)[C@@H]2O)O)O[C@H](CO)[C@H]1O DBTMGCOVALSLOR-AKJQSPAISA-N 0.000 description 4
- 235000014633 carbohydrates Nutrition 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- QIGJYVCQYDKYDW-LCOYTZNXSA-N laminarabiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@H](O)[C@@H](CO)OC(O)[C@@H]1O QIGJYVCQYDKYDW-LCOYTZNXSA-N 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 235000002639 sodium chloride Nutrition 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 108010061238 threonyl-glycine Proteins 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 229920002558 Curdlan Polymers 0.000 description 3
- 239000001879 Curdlan Substances 0.000 description 3
- DXVOKNVIKORTHQ-GUBZILKMSA-N Glu-Pro-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O DXVOKNVIKORTHQ-GUBZILKMSA-N 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 108010047495 alanylglycine Proteins 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 235000019316 curdlan Nutrition 0.000 description 3
- 229940078035 curdlan Drugs 0.000 description 3
- 108010016616 cysteinylglycine Proteins 0.000 description 3
- 150000002275 gentiobioses Chemical class 0.000 description 3
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 3
- 230000003301 hydrolyzing effect Effects 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 150000003538 tetroses Chemical class 0.000 description 3
- IVLXQGJVBGMLRR-UHFFFAOYSA-N 2-aminoacetic acid;hydron;chloride Chemical compound Cl.NCC(O)=O IVLXQGJVBGMLRR-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- HPNDBHLITCHRSO-WHFBIAKZSA-N Asp-Ala-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)NCC(O)=O HPNDBHLITCHRSO-WHFBIAKZSA-N 0.000 description 2
- 101710130006 Beta-glucanase Proteins 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 108010084185 Cellulases Proteins 0.000 description 2
- 102000005575 Cellulases Human genes 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000227647 Fucus vesiculosus Species 0.000 description 2
- VFZIDQZAEBORGY-GLLZPBPUSA-N Glu-Gln-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VFZIDQZAEBORGY-GLLZPBPUSA-N 0.000 description 2
- 108010056771 Glucosidases Proteins 0.000 description 2
- 102000004366 Glucosidases Human genes 0.000 description 2
- 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 2
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 2
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 2
- SKICPQLTOXGWGO-GARJFASQSA-N Pro-Gln-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCC(=O)N)C(=O)N2CCC[C@@H]2C(=O)O SKICPQLTOXGWGO-GARJFASQSA-N 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- FQPQPTHMHZKGFM-XQXXSGGOSA-N Thr-Ala-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O FQPQPTHMHZKGFM-XQXXSGGOSA-N 0.000 description 2
- LVHHEVGYAZGXDE-KDXUFGMBSA-N Thr-Ala-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(=O)O)N)O LVHHEVGYAZGXDE-KDXUFGMBSA-N 0.000 description 2
- CEKSLIVSNNGOKH-KZVJFYERSA-N Val-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](C(C)C)N)O CEKSLIVSNNGOKH-KZVJFYERSA-N 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 108010048056 beta-1,3-exoglucanase Proteins 0.000 description 2
- AFYNADDZULBEJA-UHFFFAOYSA-N bicinchoninic acid Chemical compound C1=CC=CC2=NC(C=3C=C(C4=CC=CC=C4N=3)C(=O)O)=CC(C(O)=O)=C21 AFYNADDZULBEJA-UHFFFAOYSA-N 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000013467 fragmentation Methods 0.000 description 2
- 238000006062 fragmentation reaction Methods 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 125000003147 glycosyl group Chemical group 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 2
- 229930027917 kanamycin Natural products 0.000 description 2
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical class O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 2
- 230000002018 overexpression Effects 0.000 description 2
- 238000013081 phylogenetic analysis Methods 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 108091033319 polynucleotide Proteins 0.000 description 2
- 102000040430 polynucleotide Human genes 0.000 description 2
- 239000002157 polynucleotide Substances 0.000 description 2
- 238000002731 protein assay Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000036632 reaction speed Effects 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 238000010188 recombinant method Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 238000001269 time-of-flight mass spectrometry Methods 0.000 description 2
- 238000010361 transduction Methods 0.000 description 2
- 230000026683 transduction Effects 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 108010080629 tryptophan-leucine Proteins 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 229920001221 xylan Polymers 0.000 description 2
- 150000004823 xylans Chemical class 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- WXTMDXOMEHJXQO-UHFFFAOYSA-N 2,5-dihydroxybenzoic acid Chemical class OC(=O)C1=CC(O)=CC=C1O WXTMDXOMEHJXQO-UHFFFAOYSA-N 0.000 description 1
- HXUVTXPOZRFMOY-NSHDSACASA-N 2-[[(2s)-2-[[2-[(2-aminoacetyl)amino]acetyl]amino]-3-phenylpropanoyl]amino]acetic acid Chemical compound NCC(=O)NCC(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=CC=C1 HXUVTXPOZRFMOY-NSHDSACASA-N 0.000 description 1
- CBNVOPQSNMPGDO-UHFFFAOYSA-N 2-[[2-[[1-[2-amino-3-(4-hydroxyphenyl)propanoyl]pyrrolidine-2-carbonyl]amino]acetyl]amino]-3-methylpentanoic acid Chemical compound CCC(C)C(C(O)=O)NC(=O)CNC(=O)C1CCCN1C(=O)C(N)CC1=CC=C(O)C=C1 CBNVOPQSNMPGDO-UHFFFAOYSA-N 0.000 description 1
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 1
- DECCMEWNXSNSDO-ZLUOBGJFSA-N Ala-Cys-Ala Chemical compound C[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O DECCMEWNXSNSDO-ZLUOBGJFSA-N 0.000 description 1
- FUSPCLTUKXQREV-ACZMJKKPSA-N Ala-Glu-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O FUSPCLTUKXQREV-ACZMJKKPSA-N 0.000 description 1
- NJPMYXWVWQWCSR-ACZMJKKPSA-N Ala-Glu-Asn Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O NJPMYXWVWQWCSR-ACZMJKKPSA-N 0.000 description 1
- CBCCCLMNOBLBSC-XVYDVKMFSA-N Ala-His-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O CBCCCLMNOBLBSC-XVYDVKMFSA-N 0.000 description 1
- CKLDHDOIYBVUNP-KBIXCLLPSA-N Ala-Ile-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O CKLDHDOIYBVUNP-KBIXCLLPSA-N 0.000 description 1
- DPNZTBKGAUAZQU-DLOVCJGASA-N Ala-Leu-His Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N DPNZTBKGAUAZQU-DLOVCJGASA-N 0.000 description 1
- QUIGLPSHIFPEOV-CIUDSAMLSA-N Ala-Lys-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O QUIGLPSHIFPEOV-CIUDSAMLSA-N 0.000 description 1
- BLTRAARCJYVJKV-QEJZJMRPSA-N Ala-Lys-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](Cc1ccccc1)C(O)=O BLTRAARCJYVJKV-QEJZJMRPSA-N 0.000 description 1
- DEWWPUNXRNGMQN-LPEHRKFASA-N Ala-Met-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N1CCC[C@@H]1C(=O)O)N DEWWPUNXRNGMQN-LPEHRKFASA-N 0.000 description 1
- DCVYRWFAMZFSDA-ZLUOBGJFSA-N Ala-Ser-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O DCVYRWFAMZFSDA-ZLUOBGJFSA-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
- WNHNMKOFKCHKKD-BFHQHQDPSA-N Ala-Thr-Gly Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O WNHNMKOFKCHKKD-BFHQHQDPSA-N 0.000 description 1
- IOFVWPYSRSCWHI-JXUBOQSCSA-N Ala-Thr-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C)N IOFVWPYSRSCWHI-JXUBOQSCSA-N 0.000 description 1
- KUFVXLQLDHJVOG-SHGPDSBTSA-N Ala-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C)N)O KUFVXLQLDHJVOG-SHGPDSBTSA-N 0.000 description 1
- IETUUAHKCHOQHP-KZVJFYERSA-N Ala-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@H](C)N)[C@@H](C)O)C(O)=O IETUUAHKCHOQHP-KZVJFYERSA-N 0.000 description 1
- BGGAIXWIZCIFSG-XDTLVQLUSA-N Ala-Tyr-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O BGGAIXWIZCIFSG-XDTLVQLUSA-N 0.000 description 1
- YJHKTAMKPGFJCT-NRPADANISA-N Ala-Val-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O YJHKTAMKPGFJCT-NRPADANISA-N 0.000 description 1
- NLYYHIKRBRMAJV-AEJSXWLSSA-N Ala-Val-Pro Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N NLYYHIKRBRMAJV-AEJSXWLSSA-N 0.000 description 1
- YYOVLDPHIJAOSY-DCAQKATOSA-N Arg-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N YYOVLDPHIJAOSY-DCAQKATOSA-N 0.000 description 1
- OTOXOKCIIQLMFH-KZVJFYERSA-N Arg-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N OTOXOKCIIQLMFH-KZVJFYERSA-N 0.000 description 1
- RRGPUNYIPJXJBU-GUBZILKMSA-N Arg-Asp-Met Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O RRGPUNYIPJXJBU-GUBZILKMSA-N 0.000 description 1
- ASQYTJJWAMDISW-BPUTZDHNSA-N Arg-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N ASQYTJJWAMDISW-BPUTZDHNSA-N 0.000 description 1
- PBSOQGZLPFVXPU-YUMQZZPRSA-N Arg-Glu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O PBSOQGZLPFVXPU-YUMQZZPRSA-N 0.000 description 1
- NMRHDSAOIURTNT-RWMBFGLXSA-N Arg-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N NMRHDSAOIURTNT-RWMBFGLXSA-N 0.000 description 1
- UZSQXCMNUPKLCC-FJXKBIBVSA-N Arg-Thr-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O UZSQXCMNUPKLCC-FJXKBIBVSA-N 0.000 description 1
- BRCVLJZIIFBSPF-ZLUOBGJFSA-N Asn-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)N)N BRCVLJZIIFBSPF-ZLUOBGJFSA-N 0.000 description 1
- WVCJSDCHTUTONA-FXQIFTODSA-N Asn-Asp-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WVCJSDCHTUTONA-FXQIFTODSA-N 0.000 description 1
- OKZOABJQOMAYEC-NUMRIWBASA-N Asn-Gln-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OKZOABJQOMAYEC-NUMRIWBASA-N 0.000 description 1
- IICZCLFBILYRCU-WHFBIAKZSA-N Asn-Gly-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IICZCLFBILYRCU-WHFBIAKZSA-N 0.000 description 1
- FTCGGKNCJZOPNB-WHFBIAKZSA-N Asn-Gly-Ser Chemical compound NC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O FTCGGKNCJZOPNB-WHFBIAKZSA-N 0.000 description 1
- BXUHCIXDSWRSBS-CIUDSAMLSA-N Asn-Leu-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O BXUHCIXDSWRSBS-CIUDSAMLSA-N 0.000 description 1
- UWFOMGUWGPRVBW-GUBZILKMSA-N Asn-Pro-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CC(=O)N)N UWFOMGUWGPRVBW-GUBZILKMSA-N 0.000 description 1
- MYTHOBCLNIOFBL-SRVKXCTJSA-N Asn-Ser-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MYTHOBCLNIOFBL-SRVKXCTJSA-N 0.000 description 1
- BCADFFUQHIMQAA-KKHAAJSZSA-N Asn-Thr-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O BCADFFUQHIMQAA-KKHAAJSZSA-N 0.000 description 1
- UPAGTDJAORYMEC-VHWLVUOQSA-N Asn-Trp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CC(=O)N)N UPAGTDJAORYMEC-VHWLVUOQSA-N 0.000 description 1
- IPAQILGYEQFCFO-NYVOZVTQSA-N Asn-Trp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CNC4=CC=CC=C43)C(=O)O)NC(=O)[C@H](CC(=O)N)N IPAQILGYEQFCFO-NYVOZVTQSA-N 0.000 description 1
- XEGZSHSPQNDNRH-JRQIVUDYSA-N Asn-Tyr-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XEGZSHSPQNDNRH-JRQIVUDYSA-N 0.000 description 1
- JNCRAQVYJZGIOW-QSFUFRPTSA-N Asn-Val-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JNCRAQVYJZGIOW-QSFUFRPTSA-N 0.000 description 1
- KDFQZBWWPYQBEN-ZLUOBGJFSA-N Asp-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N KDFQZBWWPYQBEN-ZLUOBGJFSA-N 0.000 description 1
- PBVLJOIPOGUQQP-CIUDSAMLSA-N Asp-Ala-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O PBVLJOIPOGUQQP-CIUDSAMLSA-N 0.000 description 1
- NAPNAGZWHQHZLG-ZLUOBGJFSA-N Asp-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)O)N NAPNAGZWHQHZLG-ZLUOBGJFSA-N 0.000 description 1
- XJQRWGXKUSDEFI-ACZMJKKPSA-N Asp-Glu-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O XJQRWGXKUSDEFI-ACZMJKKPSA-N 0.000 description 1
- MFTVXYMXSAQZNL-DJFWLOJKSA-N Asp-Ile-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC(=O)O)N MFTVXYMXSAQZNL-DJFWLOJKSA-N 0.000 description 1
- OEDJQRXNDRUGEU-SRVKXCTJSA-N Asp-Leu-His Chemical compound N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)O OEDJQRXNDRUGEU-SRVKXCTJSA-N 0.000 description 1
- IVPNEDNYYYFAGI-GARJFASQSA-N Asp-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N IVPNEDNYYYFAGI-GARJFASQSA-N 0.000 description 1
- HJCGDIGVVWETRO-ZPFDUUQYSA-N Asp-Lys-Ile Chemical compound CC[C@H](C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC(O)=O)C(O)=O HJCGDIGVVWETRO-ZPFDUUQYSA-N 0.000 description 1
- WDMNFNXKGSLIOB-GUBZILKMSA-N Asp-Met-Met Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)O)N WDMNFNXKGSLIOB-GUBZILKMSA-N 0.000 description 1
- LTCKTLYKRMCFOC-KKUMJFAQSA-N Asp-Phe-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O LTCKTLYKRMCFOC-KKUMJFAQSA-N 0.000 description 1
- DRCOAZZDQRCGGP-GHCJXIJMSA-N Asp-Ser-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O DRCOAZZDQRCGGP-GHCJXIJMSA-N 0.000 description 1
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 1
- KCOPOPKJRHVGPE-AQZXSJQPSA-N Asp-Thr-Trp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O KCOPOPKJRHVGPE-AQZXSJQPSA-N 0.000 description 1
- XOASPVGNFAMYBD-WFBYXXMGSA-N Asp-Trp-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(O)=O XOASPVGNFAMYBD-WFBYXXMGSA-N 0.000 description 1
- DKQCWCQRAMAFLN-UBHSHLNASA-N Asp-Trp-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(O)=O)C(O)=O DKQCWCQRAMAFLN-UBHSHLNASA-N 0.000 description 1
- CXEFNHOVIIDHFU-IHPCNDPISA-N Asp-Trp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)[C@H](CC(=O)O)N CXEFNHOVIIDHFU-IHPCNDPISA-N 0.000 description 1
- PLNJUJGNLDSFOP-UWJYBYFXSA-N Asp-Tyr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O PLNJUJGNLDSFOP-UWJYBYFXSA-N 0.000 description 1
- PLOKOIJSGCISHE-BYULHYEWSA-N Asp-Val-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O PLOKOIJSGCISHE-BYULHYEWSA-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
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000006382 Bacillus halodurans Species 0.000 description 1
- 108700038091 Beta-glucanases Proteins 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- GRNOCLDFUNCIDW-ACZMJKKPSA-N Cys-Ala-Glu Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CS)N GRNOCLDFUNCIDW-ACZMJKKPSA-N 0.000 description 1
- UXUSHQYYQCZWET-WDSKDSINSA-N Cys-Glu-Gly Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O UXUSHQYYQCZWET-WDSKDSINSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 101000874334 Dalbergia nigrescens Isoflavonoid 7-O-beta-apiosyl-glucoside beta-glycosidase Proteins 0.000 description 1
- 101710156496 Endoglucanase A Proteins 0.000 description 1
- 101000757733 Enterococcus faecalis (strain ATCC 700802 / V583) Autolysin Proteins 0.000 description 1
- 241000195620 Euglena Species 0.000 description 1
- 241000195619 Euglena gracilis Species 0.000 description 1
- 101710112457 Exoglucanase Proteins 0.000 description 1
- MFLMFRZBAJSGHK-ACZMJKKPSA-N Gln-Cys-Ser Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(=O)O)N MFLMFRZBAJSGHK-ACZMJKKPSA-N 0.000 description 1
- BLOXULLYFRGYKZ-GUBZILKMSA-N Gln-Glu-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O BLOXULLYFRGYKZ-GUBZILKMSA-N 0.000 description 1
- LFIVHGMKWFGUGK-IHRRRGAJSA-N Gln-Glu-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCC(=O)N)N LFIVHGMKWFGUGK-IHRRRGAJSA-N 0.000 description 1
- ZNTDJIMJKNNSLR-RWRJDSDZSA-N Gln-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](CCC(=O)N)N ZNTDJIMJKNNSLR-RWRJDSDZSA-N 0.000 description 1
- AKDOUBMVLRCHBD-SIUGBPQLSA-N Gln-Tyr-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O AKDOUBMVLRCHBD-SIUGBPQLSA-N 0.000 description 1
- ZZLDMBMFKZFQMU-NRPADANISA-N Gln-Val-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O ZZLDMBMFKZFQMU-NRPADANISA-N 0.000 description 1
- KBKGRMNVKPSQIF-XDTLVQLUSA-N Glu-Ala-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KBKGRMNVKPSQIF-XDTLVQLUSA-N 0.000 description 1
- NLKVNZUFDPWPNL-YUMQZZPRSA-N Glu-Arg-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O NLKVNZUFDPWPNL-YUMQZZPRSA-N 0.000 description 1
- ZOXBSICWUDAOHX-GUBZILKMSA-N Glu-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCC(O)=O ZOXBSICWUDAOHX-GUBZILKMSA-N 0.000 description 1
- PXHABOCPJVTGEK-BQBZGAKWSA-N Glu-Gln-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O PXHABOCPJVTGEK-BQBZGAKWSA-N 0.000 description 1
- KASDBWKLWJKTLJ-GUBZILKMSA-N Glu-Glu-Met Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O KASDBWKLWJKTLJ-GUBZILKMSA-N 0.000 description 1
- AAJHGGDRKHYSDH-GUBZILKMSA-N Glu-Pro-Gln Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCC(=O)O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O AAJHGGDRKHYSDH-GUBZILKMSA-N 0.000 description 1
- CQAHWYDHKUWYIX-YUMQZZPRSA-N Glu-Pro-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O CQAHWYDHKUWYIX-YUMQZZPRSA-N 0.000 description 1
- JWNZHMSRZXXGTM-XKBZYTNZSA-N Glu-Ser-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JWNZHMSRZXXGTM-XKBZYTNZSA-N 0.000 description 1
- VIPDPMHGICREIS-GVXVVHGQSA-N Glu-Val-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O VIPDPMHGICREIS-GVXVVHGQSA-N 0.000 description 1
- GZUKEVBTYNNUQF-WDSKDSINSA-N Gly-Ala-Gln Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(O)=O GZUKEVBTYNNUQF-WDSKDSINSA-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
- QXPRJQPCFXMCIY-NKWVEPMBSA-N Gly-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN QXPRJQPCFXMCIY-NKWVEPMBSA-N 0.000 description 1
- LJPIRKICOISLKN-WHFBIAKZSA-N Gly-Ala-Ser Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O LJPIRKICOISLKN-WHFBIAKZSA-N 0.000 description 1
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 1
- SOEATRRYCIPEHA-BQBZGAKWSA-N Gly-Glu-Glu Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O SOEATRRYCIPEHA-BQBZGAKWSA-N 0.000 description 1
- JUBDONGMHASUCN-IUCAKERBSA-N Gly-Glu-His Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O JUBDONGMHASUCN-IUCAKERBSA-N 0.000 description 1
- QPTNELDXWKRIFX-YFKPBYRVSA-N Gly-Gly-Gln Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCC(N)=O QPTNELDXWKRIFX-YFKPBYRVSA-N 0.000 description 1
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 1
- UQJNXZSSGQIPIQ-FBCQKBJTSA-N Gly-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)CN UQJNXZSSGQIPIQ-FBCQKBJTSA-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
- UHPAZODVFFYEEL-QWRGUYRKSA-N Gly-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN UHPAZODVFFYEEL-QWRGUYRKSA-N 0.000 description 1
- IBYOLNARKHMLBG-WHOFXGATSA-N Gly-Phe-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IBYOLNARKHMLBG-WHOFXGATSA-N 0.000 description 1
- GLACUWHUYFBSPJ-FJXKBIBVSA-N Gly-Pro-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN GLACUWHUYFBSPJ-FJXKBIBVSA-N 0.000 description 1
- OHUKZZYSJBKFRR-WHFBIAKZSA-N Gly-Ser-Asp Chemical compound [H]NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O OHUKZZYSJBKFRR-WHFBIAKZSA-N 0.000 description 1
- GULGDABMYTYMJZ-STQMWFEESA-N Gly-Trp-Asp Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(O)=O)C(O)=O GULGDABMYTYMJZ-STQMWFEESA-N 0.000 description 1
- PYFHPYDQHCEVIT-KBPBESRZSA-N Gly-Trp-Gln Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(N)=O)C(O)=O PYFHPYDQHCEVIT-KBPBESRZSA-N 0.000 description 1
- NIOPEYHPOBWLQO-KBPBESRZSA-N Gly-Trp-Glu Chemical compound NCC(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCC(O)=O)C(O)=O NIOPEYHPOBWLQO-KBPBESRZSA-N 0.000 description 1
- YDIDLLVFCYSXNY-RCOVLWMOSA-N Gly-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)CN YDIDLLVFCYSXNY-RCOVLWMOSA-N 0.000 description 1
- MUGLKCQHTUFLGF-WPRPVWTQSA-N Gly-Val-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)CN MUGLKCQHTUFLGF-WPRPVWTQSA-N 0.000 description 1
- SBVMXEZQJVUARN-XPUUQOCRSA-N Gly-Val-Ser Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O SBVMXEZQJVUARN-XPUUQOCRSA-N 0.000 description 1
- ZNPRMNDAFQKATM-LKTVYLICSA-N His-Ala-Tyr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZNPRMNDAFQKATM-LKTVYLICSA-N 0.000 description 1
- ZHHLTWUOWXHVQJ-YUMQZZPRSA-N His-Ser-Gly Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZHHLTWUOWXHVQJ-YUMQZZPRSA-N 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- HDOYNXLPTRQLAD-JBDRJPRFSA-N Ile-Ala-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)O)N HDOYNXLPTRQLAD-JBDRJPRFSA-N 0.000 description 1
- FVEWRQXNISSYFO-ZPFDUUQYSA-N Ile-Arg-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N FVEWRQXNISSYFO-ZPFDUUQYSA-N 0.000 description 1
- YPQDTQJBOFOTJQ-SXTJYALSSA-N Ile-Asn-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N YPQDTQJBOFOTJQ-SXTJYALSSA-N 0.000 description 1
- NKRJALPCDNXULF-BYULHYEWSA-N Ile-Asp-Gly Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O NKRJALPCDNXULF-BYULHYEWSA-N 0.000 description 1
- BEWFWZRGBDVXRP-PEFMBERDSA-N Ile-Glu-Asn Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O BEWFWZRGBDVXRP-PEFMBERDSA-N 0.000 description 1
- LGMUPVWZEYYUMU-YVNDNENWSA-N Ile-Glu-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N LGMUPVWZEYYUMU-YVNDNENWSA-N 0.000 description 1
- PHIXPNQDGGILMP-YVNDNENWSA-N Ile-Glu-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N PHIXPNQDGGILMP-YVNDNENWSA-N 0.000 description 1
- LPFBXFILACZHIB-LAEOZQHASA-N Ile-Gly-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)O)C(=O)O)N LPFBXFILACZHIB-LAEOZQHASA-N 0.000 description 1
- UAQSZXGJGLHMNV-XEGUGMAKSA-N Ile-Gly-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N UAQSZXGJGLHMNV-XEGUGMAKSA-N 0.000 description 1
- VNDQNDYEPSXHLU-JUKXBJQTSA-N Ile-His-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N VNDQNDYEPSXHLU-JUKXBJQTSA-N 0.000 description 1
- PWDSHAAAFXISLE-SXTJYALSSA-N Ile-Ile-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O PWDSHAAAFXISLE-SXTJYALSSA-N 0.000 description 1
- TWPSALMCEHCIOY-YTFOTSKYSA-N Ile-Ile-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)O)N TWPSALMCEHCIOY-YTFOTSKYSA-N 0.000 description 1
- QZZIBQZLWBOOJH-PEDHHIEDSA-N Ile-Ile-Val Chemical compound N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(=O)O QZZIBQZLWBOOJH-PEDHHIEDSA-N 0.000 description 1
- XLXPYSDGMXTTNQ-DKIMLUQUSA-N Ile-Phe-Leu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CC(C)C)C(O)=O XLXPYSDGMXTTNQ-DKIMLUQUSA-N 0.000 description 1
- XLXPYSDGMXTTNQ-UHFFFAOYSA-N Ile-Phe-Leu Natural products CCC(C)C(N)C(=O)NC(C(=O)NC(CC(C)C)C(O)=O)CC1=CC=CC=C1 XLXPYSDGMXTTNQ-UHFFFAOYSA-N 0.000 description 1
- ZLFNNVATRMCAKN-ZKWXMUAHSA-N Ile-Ser-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZLFNNVATRMCAKN-ZKWXMUAHSA-N 0.000 description 1
- JDCQDJVYUXNCGF-SPOWBLRKSA-N Ile-Ser-Trp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N JDCQDJVYUXNCGF-SPOWBLRKSA-N 0.000 description 1
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 1
- TYYLDKGBCJGJGW-UHFFFAOYSA-N L-tryptophan-L-tyrosine Natural products C=1NC2=CC=CC=C2C=1CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 TYYLDKGBCJGJGW-UHFFFAOYSA-N 0.000 description 1
- 241001466453 Laminaria Species 0.000 description 1
- CQQGCWPXDHTTNF-GUBZILKMSA-N Leu-Ala-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O CQQGCWPXDHTTNF-GUBZILKMSA-N 0.000 description 1
- BQSLGJHIAGOZCD-CIUDSAMLSA-N Leu-Ala-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O BQSLGJHIAGOZCD-CIUDSAMLSA-N 0.000 description 1
- VKOAHIRLIUESLU-ULQDDVLXSA-N Leu-Arg-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O VKOAHIRLIUESLU-ULQDDVLXSA-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
- OXRLYTYUXAQTHP-YUMQZZPRSA-N Leu-Gly-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(O)=O OXRLYTYUXAQTHP-YUMQZZPRSA-N 0.000 description 1
- FMEICTQWUKNAGC-YUMQZZPRSA-N Leu-Gly-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O FMEICTQWUKNAGC-YUMQZZPRSA-N 0.000 description 1
- YFBBUHJJUXXZOF-UWVGGRQHSA-N Leu-Gly-Pro Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O YFBBUHJJUXXZOF-UWVGGRQHSA-N 0.000 description 1
- HGFGEMSVBMCFKK-MNXVOIDGSA-N Leu-Ile-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O HGFGEMSVBMCFKK-MNXVOIDGSA-N 0.000 description 1
- LXKNSJLSGPNHSK-KKUMJFAQSA-N Leu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N LXKNSJLSGPNHSK-KKUMJFAQSA-N 0.000 description 1
- RXGLHDWAZQECBI-SRVKXCTJSA-N Leu-Leu-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O RXGLHDWAZQECBI-SRVKXCTJSA-N 0.000 description 1
- BIZNDKMFQHDOIE-KKUMJFAQSA-N Leu-Phe-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(N)=O)C(O)=O)CC1=CC=CC=C1 BIZNDKMFQHDOIE-KKUMJFAQSA-N 0.000 description 1
- ZAVCJRJOQKIOJW-KKUMJFAQSA-N Leu-Phe-Asp Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(O)=O)CC1=CC=CC=C1 ZAVCJRJOQKIOJW-KKUMJFAQSA-N 0.000 description 1
- AIRUUHAOKGVJAD-JYJNAYRXSA-N Leu-Phe-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIRUUHAOKGVJAD-JYJNAYRXSA-N 0.000 description 1
- WMIOEVKKYIMVKI-DCAQKATOSA-N Leu-Pro-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O WMIOEVKKYIMVKI-DCAQKATOSA-N 0.000 description 1
- ZJZNLRVCZWUONM-JXUBOQSCSA-N Leu-Thr-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O ZJZNLRVCZWUONM-JXUBOQSCSA-N 0.000 description 1
- LINKCQUOMUDLKN-KATARQTJSA-N Leu-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(C)C)N)O LINKCQUOMUDLKN-KATARQTJSA-N 0.000 description 1
- ILDSIMPXNFWKLH-KATARQTJSA-N Leu-Thr-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ILDSIMPXNFWKLH-KATARQTJSA-N 0.000 description 1
- SNOUHRPNNCAOPI-SZMVWBNQSA-N Leu-Trp-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N SNOUHRPNNCAOPI-SZMVWBNQSA-N 0.000 description 1
- 239000006137 Luria-Bertani broth Substances 0.000 description 1
- JYXBNQOKPRQNQS-YTFOTSKYSA-N Lys-Ile-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JYXBNQOKPRQNQS-YTFOTSKYSA-N 0.000 description 1
- AIRZWUMAHCDDHR-KKUMJFAQSA-N Lys-Leu-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O AIRZWUMAHCDDHR-KKUMJFAQSA-N 0.000 description 1
- YSPZCHGIWAQVKQ-AVGNSLFASA-N Lys-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCCCN YSPZCHGIWAQVKQ-AVGNSLFASA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- CTVJSFRHUOSCQQ-DCAQKATOSA-N Met-Arg-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O CTVJSFRHUOSCQQ-DCAQKATOSA-N 0.000 description 1
- OSOLWRWQADPDIQ-DCAQKATOSA-N Met-Asp-Leu Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O OSOLWRWQADPDIQ-DCAQKATOSA-N 0.000 description 1
- XPCLRYNQMZOOFB-ULQDDVLXSA-N Met-His-Phe Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)O)N XPCLRYNQMZOOFB-ULQDDVLXSA-N 0.000 description 1
- 102000008300 Mutant Proteins Human genes 0.000 description 1
- 108010021466 Mutant Proteins Proteins 0.000 description 1
- 101000757734 Mycolicibacterium phlei 38 kDa autolysin Proteins 0.000 description 1
- PESQCPHRXOFIPX-UHFFFAOYSA-N N-L-methionyl-L-tyrosine Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 PESQCPHRXOFIPX-UHFFFAOYSA-N 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- AYRXSINWFIIFAE-UHFFFAOYSA-N O6-alpha-D-Galactopyranosyl-D-galactose Natural products OCC1OC(OCC(O)C(O)C(O)C(O)C=O)C(O)C(O)C1O AYRXSINWFIIFAE-UHFFFAOYSA-N 0.000 description 1
- VHWOBXIWBDWZHK-IHRRRGAJSA-N Phe-Arg-Asp Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](CC(O)=O)C(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 VHWOBXIWBDWZHK-IHRRRGAJSA-N 0.000 description 1
- IDUCUXTUHHIQIP-SOUVJXGZSA-N Phe-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O IDUCUXTUHHIQIP-SOUVJXGZSA-N 0.000 description 1
- JJHVFCUWLSKADD-ONGXEEELSA-N Phe-Gly-Ala Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](C)C(O)=O JJHVFCUWLSKADD-ONGXEEELSA-N 0.000 description 1
- VJLLEKDQJSMHRU-STQMWFEESA-N Phe-Gly-Met Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O VJLLEKDQJSMHRU-STQMWFEESA-N 0.000 description 1
- QPVFUAUFEBPIPT-CDMKHQONSA-N Phe-Gly-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O QPVFUAUFEBPIPT-CDMKHQONSA-N 0.000 description 1
- OVJMCXAPGFDGMG-HKUYNNGSSA-N Phe-Gly-Trp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O OVJMCXAPGFDGMG-HKUYNNGSSA-N 0.000 description 1
- NRKNYPRRWXVELC-NQCBNZPSSA-N Phe-Ile-Trp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC3=CC=CC=C3)N NRKNYPRRWXVELC-NQCBNZPSSA-N 0.000 description 1
- BPCLGWHVPVTTFM-QWRGUYRKSA-N Phe-Ser-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)NCC(O)=O BPCLGWHVPVTTFM-QWRGUYRKSA-N 0.000 description 1
- SHUFSZDAIPLZLF-BEAPCOKYSA-N Phe-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N)O SHUFSZDAIPLZLF-BEAPCOKYSA-N 0.000 description 1
- MSSXKZBDKZAHCX-UNQGMJICSA-N Phe-Thr-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O MSSXKZBDKZAHCX-UNQGMJICSA-N 0.000 description 1
- KIQUCMUULDXTAZ-HJOGWXRNSA-N Phe-Tyr-Tyr Chemical compound N[C@@H](Cc1ccccc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(O)=O KIQUCMUULDXTAZ-HJOGWXRNSA-N 0.000 description 1
- CDHURCQGUDNBMA-UBHSHLNASA-N Phe-Val-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 CDHURCQGUDNBMA-UBHSHLNASA-N 0.000 description 1
- TXPUNZXZDVJUJQ-LPEHRKFASA-N Pro-Asn-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC(=O)N)C(=O)N2CCC[C@@H]2C(=O)O TXPUNZXZDVJUJQ-LPEHRKFASA-N 0.000 description 1
- HXOLCSYHGRNXJJ-IHRRRGAJSA-N Pro-Asp-Phe Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HXOLCSYHGRNXJJ-IHRRRGAJSA-N 0.000 description 1
- WSRWHZRUOCACLJ-UWVGGRQHSA-N Pro-Gly-His Chemical compound C([C@@H](C(=O)O)NC(=O)CNC(=O)[C@H]1NCCC1)C1=CN=CN1 WSRWHZRUOCACLJ-UWVGGRQHSA-N 0.000 description 1
- UIMCLYYSUCIUJM-UWVGGRQHSA-N Pro-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 UIMCLYYSUCIUJM-UWVGGRQHSA-N 0.000 description 1
- WHNJMTHJGCEKGA-ULQDDVLXSA-N Pro-Phe-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O WHNJMTHJGCEKGA-ULQDDVLXSA-N 0.000 description 1
- 241000983742 Saccharina Species 0.000 description 1
- ZOHGLPQGEHSLPD-FXQIFTODSA-N Ser-Gln-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ZOHGLPQGEHSLPD-FXQIFTODSA-N 0.000 description 1
- HJEBZBMOTCQYDN-ACZMJKKPSA-N Ser-Glu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O HJEBZBMOTCQYDN-ACZMJKKPSA-N 0.000 description 1
- IOVHBRCQOGWAQH-ZKWXMUAHSA-N Ser-Gly-Ile Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOVHBRCQOGWAQH-ZKWXMUAHSA-N 0.000 description 1
- SFTZWNJFZYOLBD-ZDLURKLDSA-N Ser-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO SFTZWNJFZYOLBD-ZDLURKLDSA-N 0.000 description 1
- LQESNKGTTNHZPZ-GHCJXIJMSA-N Ser-Ile-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O LQESNKGTTNHZPZ-GHCJXIJMSA-N 0.000 description 1
- HBTCFCHYALPXME-HTFCKZLJSA-N Ser-Ile-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HBTCFCHYALPXME-HTFCKZLJSA-N 0.000 description 1
- NQZFFLBPNDLTPO-DLOVCJGASA-N Ser-Phe-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CO)N NQZFFLBPNDLTPO-DLOVCJGASA-N 0.000 description 1
- OZPDGESCTGGNAD-CIUDSAMLSA-N Ser-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CO OZPDGESCTGGNAD-CIUDSAMLSA-N 0.000 description 1
- XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 description 1
- PURRNJBBXDDWLX-ZDLURKLDSA-N Ser-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CO)N)O PURRNJBBXDDWLX-ZDLURKLDSA-N 0.000 description 1
- FGBLCMLXHRPVOF-IHRRRGAJSA-N Ser-Tyr-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FGBLCMLXHRPVOF-IHRRRGAJSA-N 0.000 description 1
- VVKVHAOOUGNDPJ-SRVKXCTJSA-N Ser-Tyr-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O VVKVHAOOUGNDPJ-SRVKXCTJSA-N 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 241000946750 Streptomyces sioyaensis Species 0.000 description 1
- IGROJMCBGRFRGI-YTLHQDLWSA-N Thr-Ala-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O IGROJMCBGRFRGI-YTLHQDLWSA-N 0.000 description 1
- VOGXLRKCWFLJBY-HSHDSVGOSA-N Thr-Arg-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O VOGXLRKCWFLJBY-HSHDSVGOSA-N 0.000 description 1
- UNURFMVMXLENAZ-KJEVXHAQSA-N Thr-Arg-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O UNURFMVMXLENAZ-KJEVXHAQSA-N 0.000 description 1
- JEDIEMIJYSRUBB-FOHZUACHSA-N Thr-Asp-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O JEDIEMIJYSRUBB-FOHZUACHSA-N 0.000 description 1
- CQNFRKAKGDSJFR-NUMRIWBASA-N Thr-Glu-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O CQNFRKAKGDSJFR-NUMRIWBASA-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
- YSXYEJWDHBCTDJ-DVJZZOLTSA-N Thr-Gly-Trp Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O YSXYEJWDHBCTDJ-DVJZZOLTSA-N 0.000 description 1
- CRZNCABIJLRFKZ-IUKAMOBKSA-N Thr-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N CRZNCABIJLRFKZ-IUKAMOBKSA-N 0.000 description 1
- GXUWHVZYDAHFSV-FLBSBUHZSA-N Thr-Ile-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GXUWHVZYDAHFSV-FLBSBUHZSA-N 0.000 description 1
- AMXMBCAXAZUCFA-RHYQMDGZSA-N Thr-Leu-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AMXMBCAXAZUCFA-RHYQMDGZSA-N 0.000 description 1
- RRRRCRYTLZVCEN-HJGDQZAQSA-N Thr-Leu-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O RRRRCRYTLZVCEN-HJGDQZAQSA-N 0.000 description 1
- MEJHFIOYJHTWMK-VOAKCMCISA-N Thr-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)[C@@H](C)O MEJHFIOYJHTWMK-VOAKCMCISA-N 0.000 description 1
- PRNGXSILMXSWQQ-OEAJRASXSA-N Thr-Leu-Phe Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PRNGXSILMXSWQQ-OEAJRASXSA-N 0.000 description 1
- QFCQNHITJPRQTB-IEGACIPQSA-N Thr-Lys-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O QFCQNHITJPRQTB-IEGACIPQSA-N 0.000 description 1
- NWECYMJLJGCBOD-UNQGMJICSA-N Thr-Phe-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O NWECYMJLJGCBOD-UNQGMJICSA-N 0.000 description 1
- YGZWVPBHYABGLT-KJEVXHAQSA-N Thr-Pro-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 YGZWVPBHYABGLT-KJEVXHAQSA-N 0.000 description 1
- SGAOHNPSEPVAFP-ZDLURKLDSA-N Thr-Ser-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SGAOHNPSEPVAFP-ZDLURKLDSA-N 0.000 description 1
- YRJOLUDFVAUXLI-GSSVUCPTSA-N Thr-Thr-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(O)=O YRJOLUDFVAUXLI-GSSVUCPTSA-N 0.000 description 1
- ZESGVALRVJIVLZ-VFCFLDTKSA-N Thr-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@@H]1C(=O)O)N)O ZESGVALRVJIVLZ-VFCFLDTKSA-N 0.000 description 1
- NJGMALCNYAMYCB-JRQIVUDYSA-N Thr-Tyr-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O NJGMALCNYAMYCB-JRQIVUDYSA-N 0.000 description 1
- -1 Tris-HCl Polysaccharide Chemical class 0.000 description 1
- MQVGIFJSFFVGFW-XEGUGMAKSA-N Trp-Ala-Glu Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O MQVGIFJSFFVGFW-XEGUGMAKSA-N 0.000 description 1
- RYXOUTORDIUWNI-BPUTZDHNSA-N Trp-Asn-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N RYXOUTORDIUWNI-BPUTZDHNSA-N 0.000 description 1
- GWQUSADRQCTMHN-NWLDYVSISA-N Trp-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N)O GWQUSADRQCTMHN-NWLDYVSISA-N 0.000 description 1
- IJRXQJVGFBSKIV-ZFWWWQNUSA-N Trp-Gly-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC1=CNC2=CC=CC=C21)N IJRXQJVGFBSKIV-ZFWWWQNUSA-N 0.000 description 1
- CCZXBOFIBYQLEV-IHPCNDPISA-N Trp-Leu-Leu Chemical compound CC(C)C[C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)Cc1c[nH]c2ccccc12)C(O)=O CCZXBOFIBYQLEV-IHPCNDPISA-N 0.000 description 1
- ABEVJDLMFPTGPS-SZMVWBNQSA-N Trp-Met-Met Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCSC)C(O)=O ABEVJDLMFPTGPS-SZMVWBNQSA-N 0.000 description 1
- ABRICLFKFRFDKS-IHPCNDPISA-N Trp-Ser-Tyr Chemical compound C([C@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)N)C(O)=O)C1=CC=C(O)C=C1 ABRICLFKFRFDKS-IHPCNDPISA-N 0.000 description 1
- SEXRBCGSZRCIPE-LYSGOOTNSA-N Trp-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N)O SEXRBCGSZRCIPE-LYSGOOTNSA-N 0.000 description 1
- PCXFIOFKIMNHGR-UHFFFAOYSA-N Tyr Trp Gly Ser Chemical compound C=1NC2=CC=CC=C2C=1CC(C(=O)NCC(=O)NC(CO)C(O)=O)NC(=O)C(N)CC1=CC=C(O)C=C1 PCXFIOFKIMNHGR-UHFFFAOYSA-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
- IIJWXEUNETVJPV-IHRRRGAJSA-N Tyr-Arg-Ser Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N)O IIJWXEUNETVJPV-IHRRRGAJSA-N 0.000 description 1
- JRXKIVGWMMIIOF-YDHLFZDLSA-N Tyr-Asn-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CC=C(C=C1)O)N JRXKIVGWMMIIOF-YDHLFZDLSA-N 0.000 description 1
- MNMYOSZWCKYEDI-JRQIVUDYSA-N Tyr-Asp-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MNMYOSZWCKYEDI-JRQIVUDYSA-N 0.000 description 1
- ZRPLVTZTKPPSBT-AVGNSLFASA-N Tyr-Glu-Ser 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](CO)C(O)=O ZRPLVTZTKPPSBT-AVGNSLFASA-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
- HZDQUVQEVVYDDA-ACRUOGEOSA-N Tyr-Tyr-Leu Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HZDQUVQEVVYDDA-ACRUOGEOSA-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
- AUMNPAUHKUNHHN-BYULHYEWSA-N Val-Asn-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N AUMNPAUHKUNHHN-BYULHYEWSA-N 0.000 description 1
- ISERLACIZUGCDX-ZKWXMUAHSA-N Val-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C(C)C)N ISERLACIZUGCDX-ZKWXMUAHSA-N 0.000 description 1
- PWRITNSESKQTPW-NRPADANISA-N Val-Gln-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N PWRITNSESKQTPW-NRPADANISA-N 0.000 description 1
- XGJLNBNZNMVJRS-NRPADANISA-N Val-Glu-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O XGJLNBNZNMVJRS-NRPADANISA-N 0.000 description 1
- SZTTYWIUCGSURQ-AUTRQRHGSA-N Val-Glu-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O SZTTYWIUCGSURQ-AUTRQRHGSA-N 0.000 description 1
- KVRLNEILGGVBJX-IHRRRGAJSA-N Val-His-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CN=CN1 KVRLNEILGGVBJX-IHRRRGAJSA-N 0.000 description 1
- FTKXYXACXYOHND-XUXIUFHCSA-N Val-Ile-Leu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O FTKXYXACXYOHND-XUXIUFHCSA-N 0.000 description 1
- SDUBQHUJJWQTEU-XUXIUFHCSA-N Val-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](C(C)C)N SDUBQHUJJWQTEU-XUXIUFHCSA-N 0.000 description 1
- MHHAWNPHDLCPLF-ULQDDVLXSA-N Val-Phe-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CC=CC=C1 MHHAWNPHDLCPLF-ULQDDVLXSA-N 0.000 description 1
- MIKHIIQMRFYVOR-RCWTZXSCSA-N Val-Pro-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C(C)C)N)O MIKHIIQMRFYVOR-RCWTZXSCSA-N 0.000 description 1
- PQSNETRGCRUOGP-KKHAAJSZSA-N Val-Thr-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(N)=O PQSNETRGCRUOGP-KKHAAJSZSA-N 0.000 description 1
- DLRZGNXCXUGIDG-KKHAAJSZSA-N Val-Thr-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N)O DLRZGNXCXUGIDG-KKHAAJSZSA-N 0.000 description 1
- YQYFYUSYEDNLSD-YEPSODPASA-N Val-Thr-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O YQYFYUSYEDNLSD-YEPSODPASA-N 0.000 description 1
- 229920002522 Wood fibre Polymers 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010306 acid treatment Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010078114 alanyl-tryptophyl-alanine Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 108010018691 arginyl-threonyl-arginine Proteins 0.000 description 1
- 108010010430 asparagine-proline-alanine Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000002551 biofuel Substances 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 108010089934 carbohydrase Proteins 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 229940105329 carboxymethylcellulose Drugs 0.000 description 1
- 108020001778 catalytic domains Proteins 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000003245 coal Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- IJKVHSBPTUYDLN-UHFFFAOYSA-N dihydroxy(oxo)silane Chemical compound O[Si](O)=O IJKVHSBPTUYDLN-UHFFFAOYSA-N 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
- 239000012153 distilled water Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000002803 fossil fuel Substances 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- DLRVVLDZNNYCBX-CQUJWQHSSA-N gentiobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)C(O)O1 DLRVVLDZNNYCBX-CQUJWQHSSA-N 0.000 description 1
- 108010052221 glucan synthase Proteins 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 1
- 108010082286 glycyl-seryl-alanine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 150000002600 laminaribioses Chemical class 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010000761 leucylarginine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 229920005610 lignin Polymers 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 108010003700 lysyl aspartic acid Proteins 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 108010064486 phenylalanyl-leucyl-valine Proteins 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 210000002706 plastid Anatomy 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 230000007026 protein scission Effects 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000002787 reinforcement Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 108010038196 saccharide-binding proteins Proteins 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- CIJQGPVMMRXSQW-UHFFFAOYSA-M sodium;2-aminoacetic acid;hydroxide Chemical compound O.[Na+].NCC([O-])=O CIJQGPVMMRXSQW-UHFFFAOYSA-M 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000010421 standard material Substances 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- 230000017105 transposition Effects 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010020532 tyrosyl-proline Proteins 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 230000035899 viability Effects 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)
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
-
- 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)
- C12N9/2405—Glucanases
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/20—Preparation of compounds containing saccharide radicals produced by the action of an exo-1,4 alpha-glucosidase, e.g. dextrose
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01039—Glucan endo-1,3-beta-D-glucosidase (3.2.1.39)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01075—Glucan endo-1,6-beta-glucosidase (3.2.1.75)
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
本发明涉及一种由β‑葡聚糖产生的新型β‑1,3‑1,6‑内切葡聚糖酶,并产生低聚糖或葡糖。更具体地说,本发明提供一种通过使用对β‑葡聚糖展现β‑1,3‑内切葡聚糖酶活性和β‑1,6‑内切葡聚糖酶活性以及对海带低聚糖展现转糖基化活性的β‑1,3‑1,6‑内切葡聚糖酶而以高产率产生不同聚合度的低聚糖或葡糖的效应。
Description
技术领域
本发明涉及一种由β-葡聚糖产生的新型β-1,3-1,6-内切葡聚糖酶,并产生低聚糖或葡糖。
背景技术
当前,由于全球石油资源的消耗所导致的增加的石油价格及根据《联合国气候变化框架公约(the United Nations Framework Convention on Climate Change,UNFCCC)》的加强而加强环境法规,因此关于开发能够代替例如石油、煤等的常规化石燃料的生态友好型环保能源的研究正在积极进展。其中,由多种多糖组成的褐藻比木质纤维及草本生物质生长的更快且具有每培育面积的高生产率的优点。另外,由于褐藻具有较低木质素含量,因此容易转化为用于产生生物能源和化学品的原材料,且由于空气中的二氧化碳通过光合作用吸收,因此褐藻作为用于产生环境友好和经济的生物能源及化学品的资源而备受关注。
褐藻包括藻酸盐、褐藻糖胶、甘露醇、海带多糖等等,且根据褐藻的种类,褐藻衍生的多糖含有极高比率的海带多糖。举例来说,海带(Laminaria hyperborean)及糖精(Saccharina latissimi)含有呈其干重的大约32%的海带多糖,且墨角藻(Fucusvesiculosus)中总碳水化合物的大约85%是海带多糖。海带多糖具有主链,所述主链主要具有β-1,3-糖苷键和一些β-1,6-糖苷键。对于海带多糖的有效糖化,需要糖苷水解酶(glycoside hydrolases(GHs))来裂解糖苷键,且β-1,3-葡聚糖酶根据酶性质分类成两大类。β-1,3-内切葡聚糖酶裂解内部β-1,3糖苷键,由此产生多种低聚糖,且已知β-1,3-外切葡聚糖酶从非还原端产生葡糖。通过此酶反应产生的最终产物葡糖可以用作适用于产生例如乙醇等的生物燃料的原料,且通过酶反应产生的海带低聚糖也可以用作有用的生理上活性材料。
就β-1,3-内切葡聚糖酶来说,韩国专利第10-1483182号公开海带多糖通过β-1,3-内切葡聚糖酶降解成葡糖和海带二糖,但并未公开酶相对于石耳素的反应性。
发明内容
技术问题
本发明针对提供可由β-葡聚糖产生低聚糖或葡糖的新型β-1,3-1,6-内切葡聚糖酶的用途。
技术解决方案
为实现目标,本发明提供一种用于产生低聚糖或葡糖的组合物,所述组合物包含由SEQ ID NO:1中所阐述的氨基酸序列表示的β-1,3-1,6-内切葡聚糖酶(β-1,3-1,6-endoglucanase),且β-1,3-1,6-内切葡聚糖酶包含选自由以下组成的群组中的一或多种作为底物:海带多糖(laminarin)、石耳素(pustulan)以及海带低聚糖(laminarioligosaecharide)。
本发明还提供一种产生低聚糖或葡糖的方法,所述方法包含使由SEQ ID NO:1中所阐述的氨基酸序列表示的β-1,3-1,6-内切葡聚糖酶(β-1,3-1,6-endoglueanase)与一或多种底物反应,所述一或多种底物选自由以下组成的群组:海带多糖(laminarin)、石耳素(pustulan)以及海带低聚糖(laminarioligosaccharide)。
有利效应
存在本发明提供一种对于β-葡聚糖展现β-1,6-内切葡聚糖酶活性以及β-1,3-内切葡聚糖酶活性且对于对于海带低聚糖展现转糖基化活性的β-1,3-1,6-内切葡聚糖酶的效应。
β-1,3-1,6-内切葡聚糖酶可使用海带多糖、石耳素以及海带低聚糖作为底物,且由此提供以高产率产生具有多种聚合度或葡糖的低聚糖的效应。
附图说明
图1是鉴别根据本发明的β-1,3-1,6-内切葡聚糖酶的表达的凝胶图像。
图2是鉴别用于本发明的β-1,3-1,6-内切葡聚糖酶的最优活性pH的结果。
图3是鉴别用于本发明的β-1,3-1,6-内切葡聚糖酶的最优活性温度的结果。
图4是确认本发明的β-1,3-1,6-内切葡聚糖酶的热稳定性的结果。
图5示出本发明的β-1,3-1,6-内切葡聚糖酶相对于作为底物的海带多糖的水解的利-伯二氏标绘图(Lineweaver-Burk plot)。
图6是使用海带多糖作为底物的本发明的β-1,3-1,6-内切葡聚糖酶的反应产物的TLC(a)和HPLC(b)分析的结果。
图7是使用石耳素作为底物的本发明的β-1,3-1,6-内切葡聚糖酶的反应产物的TLC(a)和HPLC(b)分析的结果。
图8是使用海带多糖(a)和石耳素(b)作为底物的本发明的β-1,3-1,6-内切葡聚糖酶的反应产物的MALDI-TOF/TOF MS分析的结果。
图9是使用海带低聚糖作为底物的本发明的β-1,3-1,6-内切葡聚糖酶的TLC分析的结果。
图10a-c是使用海带低聚糖作为底物的本发明的β-1,3-1,6-内切葡聚糖酶的MALDI-TOF/TOF MS分析的结果。
图11示出本发明的β-1,3-1,6-内切葡聚糖酶的系统发生树。
具体实施方式
本发明人鉴别出属于GH5族的Gly5M蛋白的β-葡聚糖酶活性,假定所述GH5族具有β-糖苷酶活性。因此,发现Gly5M蛋白裂解了作为底物的海带多糖的β-1,3-糖苷键,所述是海带多糖由β-1,3-键联的葡糖主链和β-1,6-键联的葡糖支链组成的多糖,由此产生海带低聚糖和葡糖,且裂解了底物石耳素的β-1,6-糖苷键,所述石耳素是由通过β-1,6-键键联的葡糖单体组成的多糖,由此产生龙胆低聚糖和葡糖。当海带低聚糖用作底物时,同样确认Gly5M蛋白展现转糖基化活性,且因此产生具有比海带低聚糖作为底物更高的DP值的低聚糖。由于Gly5M蛋白的各种酶性质包含属于GH5族的其它酶中并未展示的β-1,3-内切葡聚糖酶活性,因此提供先前未知的关于GH5族酶的新信息。
因此,本发明提供一种用于产生低聚糖或葡糖的组合物,其包含由SEQ ID NO:1中所阐述的氨基酸序列表示的β-1,3-1,6-内切葡聚糖酶(β-1,3-1,6-endoglucanase),其中β-1,3-1,6-内切葡聚糖酶使用选自由以下组成的群组中的一或多种作为底物:海带多糖(laminarin)、石耳素(pustulan)以及海带低聚糖(laminarioligosaccharide)。
另外,本发明还提供一种产生低聚糖或葡糖的方法,所述方法包含使由SEQ IDNO:1中所阐述的氨基酸序列表示的β-1,3-1,6-内切葡聚糖酶与一或多种底物反应,所述一或多种底物选自由以下组成的群组:海带多糖(laminarin)、石耳素(pustulan)以及海带低聚糖(laminarioligosaccharide)。
β-1,3-1,6-内切葡聚糖酶对于β-葡聚糖展现β-1,3-内切葡聚糖酶和β-1,6-内切葡聚糖酶活性,且对于海带低聚糖展现转糖基化活性。
β-1,3-1,6-内切葡聚糖酶在至多大约20℃到45℃下保持热稳定性,且对于海带多糖、海带低聚糖或石耳素展现出最优降解活性。更具体地说,可在大约30℃到40℃下展现出最优活性。
另外,含β-1,3-1,6-内切葡聚糖酶的缓冲液的最优pH可根据缓冲液的类型而变化,但可范围介于大约pH 4到pH 10,更具体地说,大约pH 4到pH 9,且最确切地说大约pH6。
β-1,3-1,6-内切葡聚糖酶可使用海带多糖、石耳素以及海带低聚糖作为底物。
用作底物的海带低聚糖更具体来说是聚合度(degree of polymerization,DP)为2到6的海带低聚糖中的任一种。举例来说,海带低聚糖可以是海带二糖(DP2)、海带三糖(DP3)、海带四糖(DP4)、海带五糖(DP5)或海带六糖(DP6)。当DP为2到6的海带低聚糖中的任一种用作底物时,由于转糖基化活性,可获得展示指示比底物的DP值更高的DP值的峰的海带低聚糖作为降解产物。
另外,酶的反应产物可为海带低聚糖、龙胆低聚糖或葡糖。
海带低聚糖可以是聚合度为2到17的海带低聚糖中的任何一种。更具体地说,可以产生聚合度为2到6的大量海带二糖、海带三糖、海带四糖、海带五糖或海带六糖。
龙胆低聚糖可以是聚合度为2到8的龙胆低聚糖中的任何一种。更具体地说,可以产生聚合度为2的大量龙胆二糖。
β-1,3-1,6-内切葡聚糖酶可衍生自变形菌株(Saccharophagus degradans)2-40T,但本发明不受其特定限制。
另外,β-1,3-1,6-内切葡聚糖酶可由与多肽的产生相关联的DNA片段(即编码基因)转录和转译,不仅包含酶的编码区的上游区域和下游区域,还包含单个编码片段之间的内含子。举例来说,β-1,3-1,6-内切葡聚糖酶可由SEQ ID NO:2中所阐述的序列转录和转译,但本发明不受其特定限制。另外,作为衍生自具有一或多个取代、缺失、易位以及添加的酶的突变体蛋白的具有低聚糖或葡糖水解活性的蛋白同样包含于本发明的酶的范围中,且蛋白优选地包含与SEQ ID NO:1中所阐述的氨基酸序列具有至少80%、85%、90%、93%、94%、95%、96%、97%、98%或99%序列一致性的氨基酸序列。
β-1,3-1,6-内切葡聚糖酶可从变形菌株(Saccharophagus degradans)2-40T培养物的上清液分离和纯化,或可使用除变形菌株(Saccharophagus degradans)2-40T以外的菌株通过基因工程重组技术,或通过人工、化学合成方法来产生和分离。
当使用重组技术时,可使用用以促进常规重组蛋白表达的因子(例如抗生素抗性基因)以及可用于亲和性柱色谱中的报导体蛋白或肽,且这种技术包含在本发明所属领域的普通技术人员可易于实施的范围中。举例来说,β-1,3-1,6-内切葡聚糖酶可从重组载体转化的宿主细胞获得,所述重组载体包括编码β-1,3-1,6-内切葡聚糖酶的基因(即,SEQ IDNO:2中所阐述的基底序列),或其培养物。宿主细胞可以是大肠杆菌(Escherichia coli),但本发明不限于此。
可在范围介于20℃到45℃的温度和范围介于5到10的pH下进行β-1,3-1,6-内切葡聚糖酶与底物之间的反应5分钟到1天。更具体地说,当海带多糖或石耳素用作底物时,可在范围介于38℃到42℃的温度和范围介于5到7的pH下进行反应5分钟到5小时。当海带低聚糖用作底物时,可在范围介于38℃到42℃的温度和范围介于5到7的pH下进行反应1小时到5小时。
酶的降解产物可依序经历硅胶色谱(其是吸附色谱)和生物凝胶P2色谱(其是凝胶渗透色谱),以分离和纯化具有大约95%的高纯度的低聚糖或葡糖。
在本文中所使用的“蛋白”和“多肽”可互换地使用。
在本发明中,表述“多肽与另一序列具有特定百分比(例如,80%、85%、90%、95%或99%)的序列一致性”意味着当将两个序列比对且进行比较时,氨基酸残基的特定百分比是相同的。可使用本领域中已知的适合软件程序来判定比对以及同源性百分比或一致性,例如文献[《现代分子生物学实验技术(CURRENT PROTOCOLS IN MOLECULAR BIOLOGY)》(F.M.奥斯贝(F.M.Ausubel)等人,(编(eds))1987年副刊(Supplement)30章节(section)7.7.18)]中所公开的方法。可在本文中使用的优选程序包含:GCG Pileup程序、FASTA(皮尔逊(Pearson)等人1988年《美国科学院院刊(Proc.Natl Acad.Sci USA)》85:2444-2448)以及BLAST(《BLAST手册(BLAST Manual)》,阿尔丘尔(Altschul)等人,国立生物技术信息中心(Natl.Cent.Biotechnol.Inf.),国立医学图书馆(Natl Lib.Med.)(NCIB NLM NIH),贝塞斯达(Bethesda),MD以及阿尔丘尔(Altschul)等人1997年NAR25:3389-3402)。另一优选比对程序是ALIGN Plus(科学和教育软件(Scientific and Educational Software),PA),优选地使用基本参数。另一可用序列软件程序是可用于序列软件包版本6.0(SequenceSoftware Package Version 6.0)(《遗传学计算机组(Genetics Computer Group)》,威斯康星大学(University of Wisconsin),威斯康星州麦迪逊(Madison,WI))中的TFASTA数据搜索程序(TFASTA Data Searching Program)。
当关于细胞、核酸、蛋白或载体使用时,本文中所使用的术语“重组”意味着细胞、核酸、蛋白或载体已通过引入异源核酸或蛋白或改变固有核酸或蛋白而被修饰,或细胞衍生自以这种方式修饰的细胞。换句话说,重组细胞表达不以细胞的固有(非重组)形式发现的基因,或可替代地,表达异常表达或从不表达的固有基因。
本文中所使用的“核酸”涵盖单链或双链DNA、RNA以及其化学修饰的形式。本文中所使用的“核酸”和“多核苷酸”可互换地使用。由于遗传密码的简并,因此一个或多个密码子可用以编码特定氨基酸,且本发明涵盖编码特定氨基酸序列的多核苷酸。
在本文中用以描述将核酸序列插入到细胞中的术语“引入”是指“转染(transfection)”、“转化”或“转导(transduction)”,且包含将核酸序列并入到真核或原核细胞中的描述。此时,核酸序列并入到细胞的基因组(例如,染色体、质粒、质体或粒线体DNA)中,且因此转化成自主复制子,或瞬时表达。
在下文中,将参考根据本发明的实例进一步详细描述本发明,但本发明的范围不限于以下实例。
[具体实施方式]
<实例1>通过克隆技术获取gly5m基因
在含有23g/L(克/升)即溶海盐(instant ocean sea salt)、50mM(毫摩尔/升)Tris-HCl、2g/L葡糖、2g/L酵母提取物以及0.5g/L氯化铵的基本培养基中在30℃下将变形菌株(Saccharophagus degradans)2-40T(ATCC 43961)培养12个小时。
使用可商购的DNA分离试剂盒(美国加利福尼亚州凯杰巴伦西亚市(Qiagen,Valencia,CA,USA))获得变形菌株(Saccharophagus degradans)2-40T(ATCC 43961)的基因组DNA。使用Solg 2×Taq PCR智能混合2(smart mix 2)(韩国大田索尔金有限公司(SolGent,Daejeon,Korea))扩增靶基因gly5m(基因库(GenBank)ID.ABD82251.1)。本文中所使用的引物是5′-GCGGGATCCATGAGAGAAAAACTACTGCGCG-3′(正向)和5′-GCGCTCGAGGTGGTGGTGGTGGTGGTGGTCAACTGCTTCAACACTCCA-3′(反向),其中的每一个具有5’端处的BamHI限制位点和XhoI限制位点。另外,为增加HisTrap柱的亲和性,添加编码组氨酸的基因的基底序列。PCR产物和pET28a+载体两个用BamHI和XhoI进行双酶切,且连接最终的DNA片段。携带gly5m的质粒转化成大肠杆菌(Escherichia coli)DH5α。
<实例2>Gly5M的过度表达和纯化
对于实例1中所获取的基因的过度表达,基因转化成蛋白表现宿主大肠杆菌(Escherichia coli)BL21(DE3)。在含有50mg/L(毫克/升)卡那霉素(kanamycin)的卢里亚-贝尔塔尼培养液(Luria-Bertani(LB)broth)(美国马里兰州斯帕克斯BD公司(BD,Sparks,MD,USA))中在37℃下培养细菌细胞直到600nm(纳米)下的吸光率达到0.6。在16℃下使用0.1mM(毫摩尔/升)IPTG诱导蛋白表达,且因此重组蛋白以水溶性类型形式表达。对于表达的Gly5M蛋白的分离,细胞通过超声波处理和离心断裂,接着通过使用HisTrap柱(HisTrapcolumn)(美国皮斯卡塔威通用电气医疗集团(GE Healthcare,Piscataway,USA))纯化上清液。使用阿弥康超离心过滤器(Amicon Ultra Centrifugal filter)(美国马萨诸塞州比尔里卡密理博公司(Millipore,Billerica,MA,USA))浓缩纯化的蛋白,且使用二喹啉甲酸(bicinchoninic acid(BCA))蛋白分析试剂盒(protein assay kit)(美国伊利诺伊州罗克福德皮尔斯公司(Pierce,Rockford,IL,USA))来测量蛋白的浓度。
使用SDS-PAGE测量表达的Gly5M的分子量为大约95kDa(千道尔顿)(图1)。
<实例3>确认Gly5M的底物特异性和阳离子效应
为确认蛋白的底物特异性,使含于20mM(毫摩尔/升)Tris-HCl(pH6.0)中的包含石耳素、海带多糖、凝胶多糖(日本大阪和光株式会社(Wako,Osaka,Japan))、羧基甲基纤维素(美国密苏里州圣路易斯市西格玛-奥德里奇公司(Sigma-Aldrich,St Louis,MO,USA))以及木聚糖(美国密苏里州圣路易斯市西格玛-奥德里奇公司(Sigma-Aldrich,St Louis,MO,USA))的1%的不同葡聚糖与10.5μM(微摩尔/升)的Gly5M蛋白在40℃下反应30分钟。通过DNS方法来定量所产生的还原糖。
如表1中所展示,Gly5M蛋白对于海带多糖展现出最高活性,且在将石耳素用作底物时,相较于在仅将海带多糖用作底物时,Gly5M蛋白展现大约55.6%的相对活性。已确认相较于使用海带多糖的情况,Gly5M蛋白质并不水解凝胶多糖,不同于其它β-葡聚糖酶,其极其类似于在凝胶多糖中高度活跃的β-1,3-葡聚糖酶的特性。同样确认Gly5M蛋白并未在Avicel中的β-1,4-糖苷键、大麦β-葡聚糖、CM-纤维素以及木聚糖处反应。
由于确认Gly5M蛋白的阳离子效应,如表2中所展示,发现Gly5M蛋白的反应性由阳离子(例如Ni2+、Cu2+、Fe2+以及Co2+)抑制。
[表1]
[表2]
<实例4>确认Gly5M的最优pH和活化温度
为调查相对于Gly5M蛋白的活性的最优pH,使含于20mM(毫摩尔/升)glycine-HCl(甘氨酸-HCl)(pH 2.0到pH 4.0)、20mM醋酸钠(pH 4.0-6.0)、20mM Tris-HCl(pH 6.0-9.0)以及20mM glyeine-NaOH(甘氨酸-NaOH)(pH 9.0-10.0)中的2%海带多糖与Gly5M蛋白反应。
图2展示在范围介于2.0-10.0的pH下的Gly5M的相对活性。确认Gly5M在pH 6.0下展现最高活性,且酶促活性在稍微小于或高于该pH的pH下大幅度减小。
为调查相对于Gly5M蛋白的活性的最优温度,使含于20mM Tris-HCl中的2%海带多糖与Gly5M反应。
图3展示在范围介于20-70℃的温度下的Gly5M的相对活性。随着温度在20℃到40℃的范围内增加,酶促活性逐渐增强且在40℃下达到其最高值。相较于在40℃下观察到的酶促活性,酶促活性从40℃的反应温度大幅度减小。
为确认Gly5M的热稳定性,相较于在40℃下反应1小时的稳定性,当在高于40℃的温度下进行反应1小时时,确认相对活性大幅度减小(图4)。因此,将Gly5M的最优反应温度判定为30℃,其应用于所有后续实验中。
<实例5>确认Gly5M的酶反应速率
为确认Gly5M蛋白相对于海带多糖的酶反应速率,使以0.45-9.1%的不同浓度含于20mM(毫摩尔/升)Tris-HCl缓冲液中的海带多糖在pH 6.0和40℃下与蛋白反应。
因此,根据利-伯二氏标绘图(Lineweaver-Burk plot)(图5),将Km值、Vmax值以及Kcat值分别地判定为10.4g/L、10.6U/mg以及16.8s-1。
<实例6>使用TLC和HPLC确认Gly5M蛋白的酶反应的特性
使用薄层色谱(thin layer chromatography(TLC))和高效液相色谱(highperformance liquid chromatography(HPLC))分析Gly5M蛋白随时间变化的酶反应的特性。
使用正丁醇∶乙酸∶水(按体积计3∶2∶2)混合溶剂系统在硅胶60盘(silica gel 60plate)(默克公司(Merck))上产生用于TLC分析的反应产物,通过用10%(v/v)硫酸处理目测,且接着在130℃下热处理5分钟。
使用配备有凝胶穿透和配体交换柱(gel permeation and ligand exchangecolumn)(KS-802;昭和(Shodex))的安捷伦1100HPLC(Agilent 1100 HPLC)(安捷伦(Agilent))进行HPLC分析,且使用折射率检测器(refractive index detector)(安捷伦(Agilent))进行检测。用于HPLC分析的溶剂是蒸馏水,流动速率(flow rate)是0.5ml/min(毫升/分钟),且将柱温设定成80℃。用于分析的标准材料包含海带二糖(DP2)、海带三糖(DP3)、海带四糖(DP4)、海带五糖(DP5)以及海带六糖(DP6),所有均购自美嘉酶公司(Megazyme)。
作为获自Gly5M蛋白与海带多糖之间的反应的产物的TLC分析的结果(图6a),确认展现不同DP值的低聚糖是在反应后5分钟或大于5分钟产生,且同样确认随时间产生对应于DP2到DP5和葡糖的具有较低DP值的大量低聚糖。
作为针对相同反应产物进行HPLC分析的结果(图6b),确认指示DP值低于DP5的峰数量随时间增加。此结果展示Gly5M蛋白随机裂解海带多糖中的β-1,3-糖苷键。
确认当Gly5M蛋白与石耳素反应时,在较早阶段,产生具有不同DP值的低聚糖,但最终产生大量龙胆二糖(DP2)和葡糖(图7a和图7b)。
<实例7>使用MALDI-TOF/TOF MS确认Gly5M蛋白的酶反应的特性
为进一步确认Gly5M蛋白的酶反应的特性,使用海带多糖、石耳素以及海带低聚糖作为底物进行基质辅助激光解吸电离-串联飞行时间质谱(matrix-assisted laserdesorption ionization-tandem time of flight mass spectrometry(MALDI-TOF/TOFMS))分析2小时。使用于分析的每一样品与溶解于50%(v/v)乙腈中的0.3μl(微升)0.01MNaCl和0.5μl的50g/L(克/升)2,5-二羟基苯甲酸盐混合,且接着点样到不锈钢目标盘上。点样样品快速干燥以用于结晶且使用ultrafleXtreme MALDI-TOF/TOF MS system(系统)(布鲁克道尔顿(Bruker Daltonics))进行分析。使用FlexAnalysis软件(3.3版本;布鲁克道尔顿)(FlexAnalysis software(version 3.3;Bruker Daltonics))来处理原始MS数据。
当将海带多糖用作底物时,确认大部分底物通过Gly5M蛋白转化为葡糖,且除葡糖以外,主要产生范围介于DP2到DP17的不同低聚糖中的DP4和DP5(图8a)。
确认当将石耳素用作底物时,确认大部分底物转化为葡糖,且除葡糖以外,产生范围介于DP2到DP8的不同低聚糖(图8b)。
当海带低聚糖(DP2-6)用作底物时,鉴别出指示比特定底物的DP值更高的DP值的峰,且因此确认当Gly5M蛋白与海带低聚糖(DP2-6)反应时,Gly5M蛋白展现出转糖基化活性(参看图9和图10a-e)。
<实例8>Gly5M蛋白的类似性分析和系统发生分析
根据使用基因库(GenBank)中保藏的不同蛋白的推导氨基酸序列的类似性分析,Gly5M由以下组成:GH5催化域和CBM6(carbohydrate-binding module 6(碳水化合物结合模块6))域。为水解由海带多糖表示的β-1,3-葡聚糖,需要β-1,3-内切葡聚糖酶(EC3.2.1.39)和β-1,3-葡糖苷酶或β-1,3-外切葡聚糖酶(EC3.2.1.58)。在GH16、GH17、GH55、GH64、GH81以及GH128中的每一个中,β-1,3-内切葡聚糖酶在其底物中需要至少两个邻接β-1,3-糖苷键。然而,在GH5族、GH7族、GH12族、GH16族以及GH17族中,β-1,3-葡糖苷酶或β-1,3-外切葡聚糖酶(EC3.2.1.58)作用于β-1,3-葡聚糖的非还原端上以释放葡糖。GH5是最大GH族中的一个,先前被称为纤维素酶A族。到目前为止,可在CAZy数据库中获得5,000或更多个GH5酶的氨基酸序列。当前,GH5含有具有EC数值的21种以实验方式特征化的酶,其包含衍生自变形菌株(S.degradans)2-40T(表3)的19种GH5酶。GH5酶通过包含共价糖基酶中间物来共享标准机制。因此,GH5酶具有水解活性以及转糖基化活性两种。迄今为止,已知GH5族的酶展现β-1,3-内切葡聚糖酶活性。Gly5M属于GH5,且由GH5催化剂域和CBM6域组成。预测CAZy中的Gly5M是内-(1,3或1,4)-β-葡聚糖酶,且全部7个保守GH5的氨基酸残基均存在于Gly5M中。类似于Gly5M的衍生变形菌株(S.degradans)2-40T的Cel5A由两个GH5催化域和三个CBM6域组成。然而,并不熟知Cel5A的底物特异性。CBM6域充当CBM(碳水化合物结合模块(earbohydrate-binding modules)),且广泛存在于纤维素酶中。然而,一些CBM通过展示特定配体结合表面来结合于海带多糖,所述表面识别1,3-键联的葡聚糖的非还原端,例如衍生自耐盐杆菌(Bacillus halodurans)C-125和盐屋链霉菌(Streptomycessioyaensis)的海带多糖酶(laminarinases)。衍生变形菌株(S.degradans)2-40T的Gly5L由GH5域和CBM6域组成,且展现如同Gly5M的β-1,3-内切葡聚糖酶活性。由于在CBM6中并未发现GH5中的任何特征化β-1,3-外切葡聚糖酶,因此CMB6将在把β-1,3-内切葡聚糖酶活性赋予GH5酶中发挥重要作用。通常,β-1,3-葡聚糖酶的共同广泛特异性是通过β-1,3-1,4-葡聚糖酶活性对β-1,3-1,4-葡聚糖的裂解作用。然而,Gly5M中未示出这种催化活性。Gly5M不仅展现β-1,3-葡聚糖酶活性且还展现β-1,6-葡聚糖酶活性,且由此展现β-1,3-葡聚糖酶的罕见广泛特异性。另外,相较于海带多糖,Gly5M展示55.6%的相对于β-1,6-葡聚糖和石耳素的相对活性。Gly5M有效地水解石耳素以产生葡糖和龙胆二糖作为主要产物。相对于β-1,6-糖苷键的Gly5M活性大大高于任何β-1,6-葡聚糖-特定β-1,6-葡聚糖酶的活性。
对于Gly5M蛋白的系统发生分析,通过获得衍生自CAZy数据库(CAZy database)(http://www.cazy.org)的先前已知微生物的β-1,3-内切葡聚糖酶的基因信息来绘制系统发生树。
如图11中所展示,Gly5M蛋白是GH5中发现的唯一β-1,3-内切葡聚糖酶,其明确展示不同于其它先前已知β-1,3-内切葡聚糖酶的系统发生。GH5酶分类成53个亚科,且Gly5M被分类为GH547。Gly5L与Gly5M具有44%类似性,且属于GH5_47。因此,表明GH5_47酶可充当相对于整个GH5展现新型活性的β-1,3-内切葡聚糖酶。
此外,已知糖苷水解酶具有转糖基活性以及水解活性。在图11中,Gly5M相对于β-1,3-低聚糖展现出转糖基化活性。葡糖从底物的非还原端中释放且接着转移到底物自身的O-3位置处。因此,底物(海带低聚糖)可充当供体和受体以便形成具有比底物更高的DP的低聚糖。出于这个原因,一些β-1,3-1,4-内切葡聚糖酶可通过相对于糖类受体来转移具有不同DP的低聚糖来充当葡聚糖合成酶。超微眼虫藻(Euglena gracilis)衍生的EgCel17A(其是β-1,3-内切葡聚糖酶)催化此类转糖基化,但最终所有中间物降解成葡糖、海带二糖以及海带三糖。Gly5M似乎将海带低聚糖识别为高DPβ-1,3-葡聚糖低聚糖的合成中的供体和受体两个。通常,低聚糖的酶催化合成是有效方法,因为其允许在无任何保护基的情况下选择性合成定义明确的低聚糖。因此,Gly5M可用作低聚糖产生催化剂。
因此,确认Gly5M充当相对于海带多糖和石耳素的β-1,3-1,6-内切葡聚糖酶,以及具有转糖基化活性。确认根据CAZy数据库,Gly5M是新型糖苷水解酶且存在于GH5中。当海带多糖用作燃料或功能性低聚糖资源时,Gly5M可被认为是用于产生葡糖和多种海带低聚糖的合适酶。
[表3]
衍生变形菌株(S.degradans)2-40T的GH5酶
工业实用性
本发明的酶可用于产生低聚糖或葡糖的领域中。
<110> 高丽大学校产学协力团
<120> 由β葡聚糖、低聚糖或葡糖产生的新型β-1,3-1,6-内切葡聚糖酶
<130> X17U13C0019
<150> KR 10-2016-0053414
<151> 2016-04-29
<160> 4
<170> KopatentIn 2.0
<210> 1
<211> 869
<212> PRT
<213> 变形菌株 2-40T(Saccharophagus degradans 2-40T)
<400> 1
Met Arg Glu Lys Leu Leu Arg Ala Leu Leu Thr Ser Ala Lys Phe Phe
1 5 10 15
Gly Ala Ser Leu Leu Leu Leu Ser Leu Phe Asn Leu Thr Ala Cys Gly
20 25 30
Gly Gly Ser Ser Gly Thr Lys Pro Val Val Glu Glu Pro Gln Pro Glu
35 40 45
Pro Gln Pro Glu Pro Glu Pro Glu Pro Glu Pro Glu Pro Glu Pro Glu
50 55 60
Pro Glu Pro Glu Pro Gln Pro Glu Pro Glu Pro Glu Pro Asp Phe Ser
65 70 75 80
Ala Leu His Thr Asp Gly Thr Lys Trp Val Asn Ala Asn Gly Asp Gln
85 90 95
Val Leu Leu Lys Gly Val Asn Leu Gly Asn Trp Leu Leu Gln Glu Phe
100 105 110
Trp Met Met Glu Gln Gly Ser Glu Asp Val Asn Asp Gln Cys Ser Leu
115 120 125
Glu Ala Val Phe Asp Asp Arg Phe Gly Phe Ala Glu Arg Glu Arg Leu
130 135 140
Met Asp Leu Phe Arg Asp Asn Trp Ile Asn Asp Arg Asp Trp Asp Ile
145 150 155 160
Ile Ala Ser Phe Gly Met Asn Val Ile Arg Leu Pro Phe Ile Trp Asn
165 170 175
Leu Ile Glu Asp Glu Asn Asn Pro Met Thr Leu Arg Asp Asp Ala Trp
180 185 190
Gln Tyr Ile Asp Tyr Ala Ile Glu Gln Ala Glu Ala Arg Asp Met Tyr
195 200 205
Val Ile Leu Asp Leu His Gly Ala Val Gly Ala Gln Gly Trp Glu His
210 215 220
His Ser Gly Cys Ala Glu Leu Asn Glu Tyr Trp Gly Ser Glu Ala Tyr
225 230 235 240
Gln Glu Arg Thr Arg Trp Leu Trp Gln Gln Val Ala Thr Arg Tyr Ala
245 250 255
Asp Arg Asp Ala Val Ala Ala Tyr Gly Val Leu Asn Glu Pro Trp Gly
260 265 270
Thr Thr Pro Glu Asn Leu Ala Val Glu Ala Ile Glu Leu Phe Asp Ala
275 280 285
Ile Arg Glu Val Asp Ala Asp Lys Ile Ile Ile Leu Pro Gly His His
290 295 300
Ser Gly Ile His Ala Tyr Pro Asn Pro Ala Thr Val Asn Gln Thr Asn
305 310 315 320
Val Ala Tyr Glu Met His Phe Tyr Pro Gly Ile Phe Gly Trp Gly Glu
325 330 335
Ile Gly Tyr Asp Val Asn Arg Asp Trp Leu Thr Cys Gly Pro Thr Gly
340 345 350
Thr Ser Gly Val Cys Glu Trp Asp Ala Arg Leu Asp Ala Leu Asp Ser
355 360 365
Pro Phe Leu Ile Gly Glu Phe Gln Pro Trp Thr Gly Leu Gly Pro Glu
370 375 380
Leu Gly Ala Gln Ile Thr Arg Ala Thr Tyr Asp Thr Tyr Ala Ser Phe
385 390 395 400
Asp Trp Ala Ser Thr Ala Trp Ser Tyr Lys Ile Ile Thr Ser Gly Gly
405 410 415
Gly Gln Gly Gly Gly Thr Trp Gly Met Val Thr Asn Glu Arg Gly Leu
420 425 430
Gly Leu Leu Ala Lys Ala Asp Thr Trp Ala Cys Ala Gly Trp Asp Ser
435 440 445
Ser Phe Ala Asn Ala Cys Gly Val Ser Arg Thr Gly Phe Thr Pro Asp
450 455 460
Arg Glu Gly Glu Gln Thr Tyr Tyr Leu Val Ile Lys Phe Gly Ala Cys
465 470 475 480
Cys Glu Gly Asn Leu Asp Ala Thr Leu Asp Ser Ile Ser Ile Ile Asp
485 490 495
Asp Val Thr Gly Glu Glu Ile Ile Val Asn Gly Gly Phe Gly Ala Gly
500 505 510
Thr Gly Trp Thr Glu Trp Tyr Glu Ser Ala Met Pro Ile Ile Asp Tyr
515 520 525
Asn Tyr Thr Gly Ala Gly Val Pro Thr Gly Ser Asp Gly Ala Val Leu
530 535 540
Arg Met Ser Gly Ala Ala Ala Ile Asn Gly Gly Val Tyr Gln Ala Ile
545 550 555 560
Thr Leu Asp Ser Ser Lys Ser Tyr Ser Phe Ser Gly Val Phe Lys Asp
565 570 575
Asn Gly Ser Ala Ser Ala Trp Ala Glu Ile Phe Leu Val Gln Ser Gln
580 585 590
Pro Val Asp Gly Ser Asp Val Leu Ala Glu Gly Pro Phe Ala Ala Val
595 600 605
Asp Phe Leu Thr Ala Pro Ile Glu Glu Ile Glu Asn Leu Phe Glu Ala
610 615 620
Phe Gly Thr Thr Pro Tyr Asp Ile His Glu Glu Met Arg Ala Ala Met
625 630 635 640
Thr Ala Glu Thr Ala Pro Thr Leu Phe Asp Leu Pro Gly Ala Pro Thr
645 650 655
Gly Val Met Leu Ala Glu Asp Ala Gly Ala Ala Thr Ile Ser Trp Thr
660 665 670
Ala Ser Gly Asp Ala Asn Val Thr Gly Tyr Asn Val Tyr Arg Ser Thr
675 680 685
Ile Ser Gly Asn Ser Tyr Thr Leu Leu Ala Glu Asn Val Thr Ala Thr
690 695 700
Thr Phe Val Asp Ser Thr Ile Asp Gly Glu Gln Thr Phe Tyr Tyr Thr
705 710 715 720
Val Thr Ala Val Thr Asp Thr Ala Glu Ser Tyr Arg Ser Gln Glu Val
725 730 735
Ala Thr Thr Phe Val Ala Val His Leu Pro Gly Lys Val Glu Ala Glu
740 745 750
Ala His Ser Asp Met Met Gly Leu Gln Thr Glu Asn Thr Thr Asp Thr
755 760 765
Gly Gly Gly Ile Asn Ile Gly Phe Ile Asp Ala Gly Asp Trp Phe Glu
770 775 780
Tyr Glu Val Thr Ile Asp Thr Ala Ala Thr Tyr Asn Ile His Tyr Arg
785 790 795 800
Leu Ala Ser Glu Pro Gly Ser Thr Gly Phe Thr Val Ser Ile Asn Asp
805 810 815
Glu Val Leu Asn Thr Val Ala Val Pro Ala Thr Gly Gly Trp Gln Thr
820 825 830
Trp Gln Thr Glu Ser Thr Thr Ile Thr Leu Pro Ala Gly Glu His Thr
835 840 845
Leu Arg Phe Asp Ala Leu Gly Gly Gln Trp Asn Met Asn Trp Trp Ser
850 855 860
Val Glu Ala Val Asp
865
<210> 2
<211> 2610
<212> DNA
<213> 变形菌株 2-40T(Saccharophagus degradans 2-40T)
<400> 2
atgagagaaa aactactgcg cgcactgtta acaagcgcca agttctttgg ggcgagctta 60
ctactgctta gcctatttaa ccttaccgcc tgtggcggag gctccagcgg cactaagccc 120
gtagtggaag agccacagcc agaaccgcag ccagaaccag aacctgaacc ggagccagaa 180
cctgaaccgg agccagaacc agaaccgcag ccagaaccag aaccagagcc cgacttctcg 240
gccctacata ccgatggcac taaatgggtt aacgccaatg gcgaccaagt gttgttaaaa 300
ggtgtgaacc tgggcaactg gctattgcaa gagttttgga tgatggagca aggctctgag 360
gatgtgaatg atcaatgctc cctcgaagct gtttttgacg accgctttgg ctttgctgag 420
cgcgagcgtc ttatggatct gttccgcgat aattggataa acgatcgcga ctgggacatt 480
atcgcctcgt tcggtatgaa cgttattcgc ctgccgttta tttggaacct aatagaagac 540
gaaaacaacc ccatgacact gcgtgacgat gcgtggcagt acattgatta cgccattgag 600
caggccgaag cccgcgacat gtatgtaatt ttagatttgc acggtgccgt aggggcacaa 660
gggtgggagc atcacagtgg ctgtgctgag cttaatgaat actggggtag cgaagcttac 720
caagagcgta cgcgctggtt gtggcagcaa gtggctacac gctatgccga ccgcgacgca 780
gtagcggctt acggcgtgct aaacgagccg tggggcacca caccagaaaa cctcgcagta 840
gaagccatcg aattattcga tgctattcgc gaagtggatg ccgacaaaat aattatttta 900
ccagggcacc actcaggtat tcacgcgtac cctaaccctg caactgtaaa ccaaaccaat 960
gttgcttacg aaatgcactt ttaccccggt atttttggtt ggggtgaaat aggctacgac 1020
gtaaaccgcg actggttaac ctgtggccca acgggcacca gcggcgtgtg cgaatgggat 1080
gcgcgcttag atgcattaga ttccccgttt ttaattggtg aatttcaacc gtggacaggc 1140
ctaggccccg aacttggtgc gcaaattaca cgtgccactt acgataccta cgcgagtttc 1200
gattgggcat ctacggcgtg gtcttacaaa attattacca gtggcggcgg tcaaggtggt 1260
ggcacatggg gcatggtaac aaacgagcgc ggtttaggtt tattggccaa agccgatact 1320
tgggcctgtg ccggttggga tagcagcttt gccaacgcat gtggtgtaag tcgcaccggt 1380
tttacgcccg atagagaagg cgagcaaacc tactatttag tgattaaatt cggcgcctgt 1440
tgcgaaggta acctcgatgc aacattagat agcatcagca ttatcgacga tgtaaccggc 1500
gaagaaataa ttgtgaatgg cggctttggt gctggtaccg gttggaccga gtggtacgaa 1560
agtgcaatgc ccattatcga ttacaactac accggtgcag gtgtgcctac gggtagcgat 1620
ggcgcggtgt tacgcatgag tggtgctgca gccattaacg gcggcgtgta ccaagctata 1680
acgttagatt ccagcaaaag ctatagcttc tctggtgtat ttaaagataa cggcagtgca 1740
agtgcatggg cagaaatatt cttagtgcaa agccagccgg ttgatggcag cgatgtatta 1800
gccgaaggcc catttgccgc ggtagatttt ttaaccgcac cgatagaaga aatagaaaat 1860
ctatttgaag cctttggcac taccccgtac gacattcacg aagaaatgcg tgcagccatg 1920
accgccgaaa cagcgccaac cttgtttgac ctgcccggcg cacctaccgg cgtaatgcta 1980
gccgaagatg caggcgcagc aacaataagt tggaccgcaa gcggcgatgc caacgtgact 2040
ggctacaatg tttatcgctc aacaatttct ggcaacagct ataccttgtt agctgaaaat 2100
gtaacagcta ccaccttcgt ggatagcacc atagatggcg agcaaacttt ctattacacc 2160
gtaacggctg taacagacac ggcagaaagc tatcgcagcc aagaggtagc cactaccttt 2220
gtagccgtgc atttacctgg caaagtagaa gcagaagctc atagcgatat gatgggctta 2280
caaaccgaaa acactaccga taccggcggc ggtattaata ttggctttat agatgccggc 2340
gattggtttg aatacgaagt aacaatcgat accgcggcga cctataacat ccactaccgc 2400
ttagcgagtg agccgggcag cacaggcttt accgtatcta taaatgatga agtgctaaat 2460
accgttgccg tacccgctac aggcggttgg caaacatggc aaaccgaaag tacaactatc 2520
accttacccg ccggcgaaca cacattgcgc tttgatgcat tgggtggcca gtggaatatg 2580
aattggtgga gtgttgaagc agttgactag 2610
<210> 3
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> gly5m: 正向引物
<400> 3
gcgggatcca tgagagaaaa actactgcgc g 31
<210> 4
<211> 48
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> gly5m: 奖励引物
<400> 4
gcgctcgagg tggtggtggt ggtggtggtc aactgcttca acactcca 48
Claims (13)
1.一种用于产生低聚糖或葡糖的组合物,包括:
β-1,3-1,6-内切葡聚糖酶(β-1,3-1,6-endoglucanase),由SEQ ID NO:1中所阐述的氨基酸序列表示,
其中所述β-1,3-1,6-内切葡聚糖酶使用选自由以下组成的群组中的一或多种作为底物:海带多糖(laminarin)、石耳素(pustulan)以及海带低聚糖(laminarioligosaeeharide)。
2.根据权利要求1所述的用于产生低聚糖或葡糖的组合物,其中所述β-1,3-1,6-内切葡聚糖酶衍生自变形菌株(Saccharophagus degradans)2-40T。
3.根据权利要求1所述的用于产生低聚糖或葡糖的组合物,其中所述β-1,3-1,6-内切葡聚糖酶从用重组载体转化的宿主细胞获得,所述重组载体包括编码所述β-1,3-1,6-内切葡聚糖酶的基因或其培养物。
4.根据权利要求3所述的用于产生低聚糖或葡糖的组合物,其中所述基因由SEQ IDNO:2中所阐述的基底序列表示。
5.根据权利要求1所述的用于产生低聚糖或葡糖的组合物,其中所述海带低聚糖是聚合度(degree ofpolymerization)为2到6的海带低聚糖中的任一种。
6.根据权利要求1所述的用于产生低聚糖或葡糖的组合物,其中通过所述β-1,3-1,6-内切葡聚糖酶产生的所述低聚糖包含海带低聚糖以及龙胆低聚糖中的任一种。
7.根据权利要求6所述的用于产生低聚糖或葡糖的组合物,其中所述海带低聚糖是聚合度为2到17的海带低聚糖中的任一种。
8.根据权利要求7所述的用于产生低聚糖或葡糖的组合物,其中所述海带低聚糖是聚合度为2到6的海带低聚糖中的任一种。
9.根据权利要求6所述的用于产生低聚糖或葡糖的组合物,其中所述龙胆低聚糖是聚合度为2到8的龙胆低聚糖中的任一种。
10.根据权利要求9所述的用于产生低聚糖或葡糖的组合物,其中所述龙胆低聚糖是龙胆二糖。
11.一种产生低聚糖或葡糖的方法,包括:
使由SEQ ID NO:1中所阐述的所述氨基酸序列表示的β-1,3-1,6-内切葡聚糖酶(β-1,3-1,6-endoglucanase)与一或多种底物反应,所述一或多种底物选自由以下组成的群组:海带多糖(laminarin)、石耳素(pustulan)以及海带低聚糖(laminarioligosaccharide)。
12.根据权利要求11所述的产生低聚糖或葡糖的方法,其中所述海带低聚糖是聚合度为2到6的海带低聚糖中的任一种。
13.根据权利要求11所述的产生低聚糖或葡糖的方法,其中所述反应是在范围介于20℃到45℃的温度以及范围介于5到10的pH下进行5分钟到1天。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020160053414A KR101804050B1 (ko) | 2016-04-29 | 2016-04-29 | 베타―글루칸으로부터 올리고당 또는 글루코스를 생산하는 신규한 베타-1,3-1,6-엔도글루카네이즈 |
KR10-2016-0053414 | 2016-04-29 | ||
PCT/KR2017/004591 WO2017188788A1 (ko) | 2016-04-29 | 2017-04-28 | 베타-글루칸으로부터 올리고당 또는 글루코스를 생산하는 신규한 베타-1,3-1,6-엔도글루카네이즈 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN109072270A true CN109072270A (zh) | 2018-12-21 |
CN109072270B CN109072270B (zh) | 2022-03-08 |
Family
ID=60159933
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201780026365.3A Active CN109072270B (zh) | 2016-04-29 | 2017-04-28 | 用于产生低聚糖或葡萄糖的组合物及其产生的方法 |
Country Status (4)
Country | Link |
---|---|
US (1) | US10724061B2 (zh) |
KR (1) | KR101804050B1 (zh) |
CN (1) | CN109072270B (zh) |
WO (1) | WO2017188788A1 (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109852647A (zh) * | 2019-03-18 | 2019-06-07 | 江南大学 | 一种双酶法制备可溶性的酵母葡聚糖寡糖的方法 |
CN111593034A (zh) * | 2020-06-24 | 2020-08-28 | 江南大学 | 利用β-1,6-葡聚糖酶制备低聚龙胆糖的方法及其应用 |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102011718B1 (ko) | 2018-03-12 | 2019-08-19 | 고려대학교 산학협력단 | 해조류로부터 포도당 및 라미나리올리고당을 생산하는 신규한 베타―글루코시데이즈 |
CN112430614B (zh) * | 2020-11-09 | 2022-10-25 | 集美大学 | 一种昆布多糖酶及其应用 |
CN116262925A (zh) * | 2021-12-13 | 2023-06-16 | 中国科学院大连化学物理研究所 | 一种β-1,3-葡聚糖酶及其制备方法和应用 |
CN115927508A (zh) * | 2022-08-29 | 2023-04-07 | 中国海洋大学 | 昆布多糖降解酶OUC-ScLam39在制备昆布寡糖中的应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20060105442A1 (en) * | 2004-11-10 | 2006-05-18 | Wu J H D | Promoters and proteins from Clostridium thermocellum and uses thereof |
CN103124783A (zh) * | 2010-06-03 | 2013-05-29 | 马斯科马公司 | 表达用于使用淀粉和纤维素进行联合生物加工的糖分解酶的酵母 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101483182B1 (ko) * | 2014-09-02 | 2015-01-19 | 한국해양과학기술원 | 신규한 엔도 베타-1,3-글루카네이즈 및 그 제조 방법 |
-
2016
- 2016-04-29 KR KR1020160053414A patent/KR101804050B1/ko active IP Right Grant
-
2017
- 2017-04-28 WO PCT/KR2017/004591 patent/WO2017188788A1/ko active Application Filing
- 2017-04-28 CN CN201780026365.3A patent/CN109072270B/zh active Active
- 2017-04-28 US US16/096,351 patent/US10724061B2/en active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20060105442A1 (en) * | 2004-11-10 | 2006-05-18 | Wu J H D | Promoters and proteins from Clostridium thermocellum and uses thereof |
CN103124783A (zh) * | 2010-06-03 | 2013-05-29 | 马斯科马公司 | 表达用于使用淀粉和纤维素进行联合生物加工的糖分解酶的酵母 |
Non-Patent Citations (5)
Title |
---|
DAMAO WANG ET AL.: ""A Novel Glycoside Hydrolase Family 5β-1,3-1,6-Endoglucanase from Saccharophagus degradans 2-40T and Its Transglycosylase Activity"", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
LIS SCHWARTZ MIOTTO ET AL.: ""The Characterization of the Endoglucanase Cel12A from Gloeophyllum trabeum Reveals an Enzyme Highly Active on β-Glucan"", 《PLOS ONE》 * |
RONAL D M. WEINER ET AL.: ""Complete Genome Sequence of the Complex Carbohydrate-Degading Marine Bacterium, Saccharophagus degradans Strain 2-40T"", 《PLOS GENET》 * |
WEINER, R.M. ET AL.: ""Saccharophagus degradans 2-40,complete genome(3860209..3862818)",Accession Number:CP000282.1", 《GENBANK》 * |
刘杰凤 等: ""海洋微生物纤维素酶的研究进展及其应用"", 《生物技术通报》 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109852647A (zh) * | 2019-03-18 | 2019-06-07 | 江南大学 | 一种双酶法制备可溶性的酵母葡聚糖寡糖的方法 |
CN111593034A (zh) * | 2020-06-24 | 2020-08-28 | 江南大学 | 利用β-1,6-葡聚糖酶制备低聚龙胆糖的方法及其应用 |
Also Published As
Publication number | Publication date |
---|---|
KR20170124149A (ko) | 2017-11-10 |
US20200002738A1 (en) | 2020-01-02 |
KR101804050B1 (ko) | 2017-12-04 |
WO2017188788A1 (ko) | 2017-11-02 |
CN109072270B (zh) | 2022-03-08 |
US10724061B2 (en) | 2020-07-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109072270A (zh) | 由β葡聚糖、低聚糖或葡糖产生的新型β-1,3-1,6-内切葡聚糖酶 | |
WO2016054176A1 (en) | Compositions comprising beta-mannanase and methods of use | |
KR20150079676A (ko) | 마그나포르테 그리세아로부터의 베타-글루코시다제 | |
Wang et al. | A novel glycoside hydrolase family 5 β-1, 3-1, 6-endoglucanase from Saccharophagus degradans 2-40T and its transglycosylase activity | |
WO2014088935A2 (en) | Compositions and methods of use | |
Aragon et al. | Production of xylo-oligosaccharides by immobilized-stabilized derivatives of endo-xylanase from Streptomyces halstedii | |
WO2014088940A1 (en) | Compositions and methods of use | |
KR101864350B1 (ko) | 베타-아가레이즈, 카파-카라기나아제 및 무수갈락토시다아제를 함유하는 효소 복합체 및 그 용도 | |
EP2929022B1 (en) | Compositions and methods of use | |
US10883128B2 (en) | Method of producing gentiobiose or glucose from β-glucan using β-1,6-endoglucanase | |
CN102719458B (zh) | 一种编码碱性β-葡萄糖苷酶基因及其应用 | |
EP2929023B1 (en) | Compositions and methods of use | |
WO2016054185A1 (en) | Compositions comprising beta-mannanase and methods of use | |
CN107699551B (zh) | 一种β-葡萄糖苷酶SPBGL5在水解木聚糖多糖类物质中的应用 | |
Brumm et al. | Identification, cloning and characterization of Dictyoglomus turgidum CelA, an endoglucanase with cellulase and mannanase activity | |
JP5777128B2 (ja) | 新規なセルラーゼ | |
KR101653340B1 (ko) | 신규의 고호열성 베타-글루코시다아제 및 이의 제조방법 | |
EP3201329A1 (en) | Compositions comprising beta mannanase and methods of use | |
US9371520B2 (en) | Fusion proteins comprising type-II cohesin modules, multi-enzyme complexes comprising same and uses thereof | |
US20170211053A1 (en) | Compositions comprising beta mannanase and methods of use | |
WO2016054205A1 (en) | Compositions comprising beta mannanase and methods of use | |
WO2016054194A1 (en) | Compositions comprising beta-mannanase and methods of use | |
KR20100119410A (ko) | 자일라네이즈 및 익스팬신을 포함하는 자일란 분해 촉진용 조성물 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |