JPWO2014157141A1 - 細胞表層発現用ポリヌクレオチド - Google Patents
細胞表層発現用ポリヌクレオチド Download PDFInfo
- Publication number
- JPWO2014157141A1 JPWO2014157141A1 JP2015508514A JP2015508514A JPWO2014157141A1 JP WO2014157141 A1 JPWO2014157141 A1 JP WO2014157141A1 JP 2015508514 A JP2015508514 A JP 2015508514A JP 2015508514 A JP2015508514 A JP 2015508514A JP WO2014157141 A1 JPWO2014157141 A1 JP WO2014157141A1
- Authority
- JP
- Japan
- Prior art keywords
- sed1
- gene
- strain
- cell surface
- yeast
- 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
- 210000004027 cell Anatomy 0.000 title claims abstract description 121
- 108091033319 polynucleotide Proteins 0.000 title claims abstract description 62
- 102000040430 polynucleotide Human genes 0.000 title claims abstract description 62
- 239000002157 polynucleotide Substances 0.000 title claims abstract description 62
- 230000014509 gene expression Effects 0.000 title claims abstract description 32
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 191
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 104
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 90
- 102000004190 Enzymes Human genes 0.000 claims abstract description 60
- 108090000790 Enzymes Proteins 0.000 claims abstract description 60
- 210000000170 cell membrane Anatomy 0.000 claims abstract description 28
- 108010076504 Protein Sorting Signals Proteins 0.000 claims abstract description 25
- 230000027455 binding Effects 0.000 claims abstract description 25
- 230000028327 secretion Effects 0.000 claims abstract description 16
- 238000004519 manufacturing process Methods 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims description 55
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 44
- 238000000855 fermentation Methods 0.000 claims description 19
- 230000004151 fermentation Effects 0.000 claims description 19
- 101150093640 SSD1 gene Proteins 0.000 claims description 18
- 239000013604 expression vector Substances 0.000 claims description 17
- 101100045541 Homo sapiens TBCD gene Proteins 0.000 claims description 12
- 101100111629 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) KAR2 gene Proteins 0.000 claims description 12
- 102100030290 Tubulin-specific chaperone D Human genes 0.000 claims description 12
- 230000002950 deficient Effects 0.000 claims description 11
- 230000001461 cytolytic effect Effects 0.000 claims description 9
- 230000003625 amylolytic effect Effects 0.000 claims description 7
- 101710191666 Lactadherin Proteins 0.000 claims 3
- 102100039648 Lactadherin Human genes 0.000 claims 3
- 101100008072 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) CWP2 gene Proteins 0.000 claims 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 77
- 230000000694 effects Effects 0.000 description 70
- 235000018102 proteins Nutrition 0.000 description 70
- 229940088598 enzyme Drugs 0.000 description 56
- 239000012634 fragment Substances 0.000 description 51
- 239000013612 plasmid Substances 0.000 description 41
- 108010047754 beta-Glucosidase Proteins 0.000 description 36
- 102000006995 beta-Glucosidase Human genes 0.000 description 36
- 108020004414 DNA Proteins 0.000 description 30
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 description 30
- 108010059892 Cellulase Proteins 0.000 description 28
- 150000001413 amino acids Chemical group 0.000 description 26
- 108010091384 endoglucanase 2 Proteins 0.000 description 26
- 108090000637 alpha-Amylases Proteins 0.000 description 24
- 239000001913 cellulose Substances 0.000 description 23
- 229920002678 cellulose Polymers 0.000 description 23
- 108091026890 Coding region Proteins 0.000 description 22
- 102100022624 Glucoamylase Human genes 0.000 description 21
- 230000003248 secreting effect Effects 0.000 description 19
- 108010008885 Cellulose 1,4-beta-Cellobiosidase Proteins 0.000 description 18
- 102000004139 alpha-Amylases Human genes 0.000 description 18
- 101150105426 BGL1 gene Proteins 0.000 description 17
- 229940024171 alpha-amylase Drugs 0.000 description 17
- 229920002472 Starch Polymers 0.000 description 16
- 239000008107 starch Substances 0.000 description 16
- 235000019698 starch Nutrition 0.000 description 16
- 241000793189 Saccharomyces cerevisiae BY4741 Species 0.000 description 15
- 239000013598 vector Substances 0.000 description 15
- 230000004186 co-expression Effects 0.000 description 13
- 230000007062 hydrolysis Effects 0.000 description 13
- 238000006460 hydrolysis reaction Methods 0.000 description 13
- 239000002028 Biomass Substances 0.000 description 12
- 101150009006 HIS3 gene Proteins 0.000 description 12
- 101100394989 Rhodopseudomonas palustris (strain ATCC BAA-98 / CGA009) hisI gene Proteins 0.000 description 12
- XIXADJRWDQXREU-UHFFFAOYSA-M lithium acetate Chemical compound [Li+].CC([O-])=O XIXADJRWDQXREU-UHFFFAOYSA-M 0.000 description 12
- 241000209094 Oryza Species 0.000 description 11
- 235000007164 Oryza sativa Nutrition 0.000 description 11
- 235000009566 rice Nutrition 0.000 description 11
- 101100437484 Arabidopsis thaliana BGLU18 gene Proteins 0.000 description 10
- 101100342633 Bos taurus LLGL1 gene Proteins 0.000 description 10
- 101100065855 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) EXG1 gene Proteins 0.000 description 10
- 101100058298 Saccharomycopsis fibuligera BGL1 gene Proteins 0.000 description 10
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 10
- 235000001014 amino acid Nutrition 0.000 description 10
- 101150100570 bglA gene Proteins 0.000 description 10
- 101150006457 sed1 gene Proteins 0.000 description 10
- 238000006467 substitution reaction Methods 0.000 description 10
- 101710098246 Exoglucanase 2 Proteins 0.000 description 9
- 229940024606 amino acid Drugs 0.000 description 9
- 230000008859 change Effects 0.000 description 9
- 239000010902 straw Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 210000005253 yeast cell Anatomy 0.000 description 9
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 8
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 8
- 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 description 8
- 230000000593 degrading effect Effects 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- 230000036962 time dependent Effects 0.000 description 8
- 101710204899 Alpha-agglutinin Proteins 0.000 description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 239000012153 distilled water Substances 0.000 description 7
- 239000008103 glucose Substances 0.000 description 7
- 239000003550 marker Substances 0.000 description 7
- 239000002773 nucleotide Substances 0.000 description 7
- 125000003729 nucleotide group Chemical group 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 239000004382 Amylase Substances 0.000 description 6
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 6
- 238000003776 cleavage reaction Methods 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 230000007017 scission Effects 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- 230000009466 transformation Effects 0.000 description 6
- 102000013142 Amylases Human genes 0.000 description 5
- 108010065511 Amylases Proteins 0.000 description 5
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 5
- 240000005384 Rhizopus oryzae Species 0.000 description 5
- 235000013752 Rhizopus oryzae Nutrition 0.000 description 5
- 241000499912 Trichoderma reesei Species 0.000 description 5
- 235000019418 amylase Nutrition 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- 229940106157 cellulase Drugs 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 229930182817 methionine Natural products 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 150000007523 nucleic acids Chemical class 0.000 description 5
- 229940035893 uracil Drugs 0.000 description 5
- 239000007222 ypd medium Substances 0.000 description 5
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 4
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 4
- 101710088194 Dehydrogenase Proteins 0.000 description 4
- 238000007792 addition Methods 0.000 description 4
- 238000002869 basic local alignment search tool Methods 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 210000000349 chromosome Anatomy 0.000 description 4
- 238000012217 deletion Methods 0.000 description 4
- 230000037430 deletion Effects 0.000 description 4
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 230000035772 mutation Effects 0.000 description 4
- 150000003905 phosphatidylinositols Chemical class 0.000 description 4
- 108091008146 restriction endonucleases Proteins 0.000 description 4
- 239000001509 sodium citrate Substances 0.000 description 4
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 4
- 241000228212 Aspergillus Species 0.000 description 3
- 240000006439 Aspergillus oryzae Species 0.000 description 3
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 3
- 101710099628 Beta-glucosidase 1 Proteins 0.000 description 3
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 3
- 102000004157 Hydrolases Human genes 0.000 description 3
- 108090000604 Hydrolases Proteins 0.000 description 3
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 3
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 241000194049 Streptococcus equinus Species 0.000 description 3
- IXKSXJFAGXLQOQ-XISFHERQSA-N WHWLQLKPGQPMY Chemical compound C([C@@H](C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)C1=CNC=N1 IXKSXJFAGXLQOQ-XISFHERQSA-N 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 210000004899 c-terminal region Anatomy 0.000 description 3
- 229940041514 candida albicans extract Drugs 0.000 description 3
- 210000002421 cell wall Anatomy 0.000 description 3
- FYGDTMLNYKFZSV-ZWSAEMDYSA-N cellotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](OC(O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-ZWSAEMDYSA-N 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 238000000354 decomposition reaction Methods 0.000 description 3
- -1 inositol phospholipid Chemical class 0.000 description 3
- 150000002632 lipids Chemical group 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 239000002344 surface layer Substances 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- 239000012138 yeast extract Substances 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- FQVLRGLGWNWPSS-BXBUPLCLSA-N (4r,7s,10s,13s,16r)-16-acetamido-13-(1h-imidazol-5-ylmethyl)-10-methyl-6,9,12,15-tetraoxo-7-propan-2-yl-1,2-dithia-5,8,11,14-tetrazacycloheptadecane-4-carboxamide Chemical compound N1C(=O)[C@@H](NC(C)=O)CSSC[C@@H](C(N)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C)NC(=O)[C@@H]1CC1=CN=CN1 FQVLRGLGWNWPSS-BXBUPLCLSA-N 0.000 description 2
- IFBHRQDFSNCLOZ-UHFFFAOYSA-N 2-(hydroxymethyl)-6-(4-nitrophenoxy)oxane-3,4,5-triol Chemical compound OC1C(O)C(O)C(CO)OC1OC1=CC=C([N+]([O-])=O)C=C1 IFBHRQDFSNCLOZ-UHFFFAOYSA-N 0.000 description 2
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 2
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 description 2
- 102100034035 Alcohol dehydrogenase 1A Human genes 0.000 description 2
- 101710191958 Amino-acid acetyltransferase Proteins 0.000 description 2
- 102000009042 Argininosuccinate Lyase Human genes 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 101150032680 CWP2 gene Proteins 0.000 description 2
- 101100032284 Candida albicans (strain SC5314 / ATCC MYA-2876) URA9 gene Proteins 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 2
- 102000048120 Galactokinases Human genes 0.000 description 2
- 108700023157 Galactokinases Proteins 0.000 description 2
- 101000892220 Geobacillus thermodenitrificans (strain NG80-2) Long-chain-alcohol dehydrogenase 1 Proteins 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 229920002527 Glycogen Polymers 0.000 description 2
- 229930186217 Glycolipid Natural products 0.000 description 2
- 101000780443 Homo sapiens Alcohol dehydrogenase 1A Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 108091092724 Noncoding DNA Proteins 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- 108020005091 Replication Origin Proteins 0.000 description 2
- 244000253724 Saccharomyces cerevisiae S288c Species 0.000 description 2
- 235000004905 Saccharomyces cerevisiae S288c Nutrition 0.000 description 2
- 108010075344 Tryptophan synthase Proteins 0.000 description 2
- 101150011703 URA1 gene Proteins 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 239000000910 agglutinin Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000013599 cloning vector Substances 0.000 description 2
- 239000000306 component Substances 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 229940096919 glycogen Drugs 0.000 description 2
- 239000002029 lignocellulosic biomass Substances 0.000 description 2
- 229920001542 oligosaccharide Polymers 0.000 description 2
- 150000002482 oligosaccharides Chemical class 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 108010085336 phosphoribosyl-AMP cyclohydrolase Proteins 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 150000004804 polysaccharides Chemical class 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 159000000000 sodium salts Chemical class 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000014616 translation Effects 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 1
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 1
- 102000005369 Aldehyde Dehydrogenase Human genes 0.000 description 1
- 108020002663 Aldehyde Dehydrogenase Proteins 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 102100033770 Alpha-amylase 1C Human genes 0.000 description 1
- 101100489349 Arabidopsis thaliana PAT24 gene Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 241000228215 Aspergillus aculeatus Species 0.000 description 1
- 241000228245 Aspergillus niger Species 0.000 description 1
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 1
- 102100030981 Beta-alanine-activating enzyme Human genes 0.000 description 1
- 101150089473 CWP1 gene Proteins 0.000 description 1
- 101100083069 Candida albicans (strain SC5314 / ATCC MYA-2876) PGA62 gene Proteins 0.000 description 1
- 101100106993 Candida albicans (strain SC5314 / ATCC MYA-2876) YWP1 gene Proteins 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- 241000186321 Cellulomonas Species 0.000 description 1
- 241000186320 Cellulomonas fimi Species 0.000 description 1
- 241000186217 Cellulomonas uda Species 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- 241000722863 Cortaderia jubata Species 0.000 description 1
- 230000004568 DNA-binding Effects 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 108010052167 Dihydroorotate Dehydrogenase Proteins 0.000 description 1
- 102100032823 Dihydroorotate dehydrogenase (quinone), mitochondrial Human genes 0.000 description 1
- 101150054379 FLO1 gene Proteins 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 229920002488 Hemicellulose Polymers 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 101000779870 Homo sapiens Alpha-amylase 1B Proteins 0.000 description 1
- 101000779869 Homo sapiens Alpha-amylase 1C Proteins 0.000 description 1
- 101000773364 Homo sapiens Beta-alanine-activating enzyme Proteins 0.000 description 1
- 101000579123 Homo sapiens Phosphoglycerate kinase 1 Proteins 0.000 description 1
- 101100315589 Homo sapiens TAX1BP3 gene Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102100033448 Lysosomal alpha-glucosidase Human genes 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108010021466 Mutant Proteins Proteins 0.000 description 1
- 102000008300 Mutant Proteins Human genes 0.000 description 1
- 101100313925 Oryza sativa subsp. japonica TIP1-2 gene Proteins 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- KJWZYMMLVHIVSU-IYCNHOCDSA-N PGK1 Chemical compound CCCCC[C@H](O)\C=C\[C@@H]1[C@@H](CCCCCCC(O)=O)C(=O)CC1=O KJWZYMMLVHIVSU-IYCNHOCDSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102100028251 Phosphoglycerate kinase 1 Human genes 0.000 description 1
- 108010086950 Phosphoribosylanthranilate isomerase Proteins 0.000 description 1
- 244000273256 Phragmites communis Species 0.000 description 1
- 235000014676 Phragmites communis Nutrition 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000589540 Pseudomonas fluorescens Species 0.000 description 1
- 241000193448 Ruminiclostridium thermocellum Species 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 101100313932 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) TIP20 gene Proteins 0.000 description 1
- 241000235344 Saccharomycetaceae Species 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- 108091058545 Secretory proteins Proteins 0.000 description 1
- 102000040739 Secretory proteins Human genes 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 102100036221 Tax1-binding protein 3 Human genes 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 102000014384 Type C Phospholipases Human genes 0.000 description 1
- 108010079194 Type C Phospholipases Proteins 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 101100313921 Zea mays TIP1-1 gene Proteins 0.000 description 1
- 108010084455 Zeocin Proteins 0.000 description 1
- HFYBTHCYPKEDQQ-UHFFFAOYSA-N [2,3-dihydroxy-3-(1h-imidazol-5-yl)propyl] dihydrogen phosphate Chemical compound OP(=O)(O)OCC(O)C(O)C1=CN=CN1 HFYBTHCYPKEDQQ-UHFFFAOYSA-N 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 239000012670 alkaline solution Substances 0.000 description 1
- 108010028144 alpha-Glucosidases Proteins 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 210000004507 artificial chromosome Anatomy 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 239000002551 biofuel Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000010432 diamond Substances 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 108010091371 endoglucanase 1 Proteins 0.000 description 1
- 108010092413 endoglucanase V Proteins 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 102000034287 fluorescent proteins Human genes 0.000 description 1
- 108091006047 fluorescent proteins Proteins 0.000 description 1
- 239000002803 fossil fuel Substances 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 238000003198 gene knock in Methods 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 229960002442 glucosamine Drugs 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 229930004094 glycosylphosphatidylinositol Natural products 0.000 description 1
- 210000002288 golgi apparatus Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 239000010903 husk Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 238000001638 lipofection Methods 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 229940049920 malate Drugs 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N malic acid Chemical compound OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 210000001322 periplasm Anatomy 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- CWCMIVBLVUHDHK-ZSNHEYEWSA-N phleomycin D1 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC[C@@H](N=1)C=1SC=C(N=1)C(=O)NCCCCNC(N)=N)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C CWCMIVBLVUHDHK-ZSNHEYEWSA-N 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000010908 plant waste Substances 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 150000004053 quinones Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000003938 response to stress Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 210000004739 secretory vesicle Anatomy 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 101150023068 tip1 gene Proteins 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000003151 transfection method Methods 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 108010087967 type I signal peptidase Proteins 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 229920001221 xylan Polymers 0.000 description 1
- 150000004823 xylans Chemical class 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
- C07K14/39—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from yeasts
-
- 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
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
-
- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
- C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
-
- 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)
- C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/035—Fusion polypeptide containing a localisation/targetting motif containing a signal for targeting to the external surface of a cell, e.g. to the outer membrane of Gram negative bacteria, GPI- anchored eukaryote proteins
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Biomedical Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
上記形質転換酵母を発酵培養する工程を含む、方法を提供する。
「細胞表層局在タンパク質」は、細胞表層に固定化または付着もしくは接着し、そこに局在するタンパク質をいう。細胞表層局在タンパク質としては、脂質で修飾されたタンパク質が知られており、この脂質が膜成分と共有結合することにより細胞膜に固定される。本明細書中では、それらの役割から、「細胞表層局在タンパク質またはその細胞膜結合領域」をまとめて、単に「アンカー」とも称する。
「分泌シグナル配列」は、分泌シグナルペプチドをコードするポリヌクレオチド配列である。
プロモーターは、プロモーター活性を有していればよく、「プロモーター活性」とは、プロモーター領域に転写因子が結合し、転写を惹起する活性をいう。プロモーターは、所望のプロモーター領域を保有する菌体、ファージなどから、制限酵素を用いて切り出し得る。必要に応じて制限酵素認識部位またはクローニングベクターとの重複部位を設けたプライマーを用い、PCRで所望のプロモーター領域を増幅することによりプロモーター領域のDNA断片を得ることができる。また、既に判明しているプロモーター領域の塩基配列情報をもとにして、所望のプロモーターを化学合成してもよい。
目的タンパク質の種類、もしくはその起源は特に限定されない。目的タンパク質の種類として、例えば、酵素、抗体、リガンド、蛍光タンパク質などが挙げられる。酵素としては、例えば、セルロース分解酵素、デンプン分解酵素、グリコーゲン分解酵素、キシラン分解酵素、キチン分解酵素、脂質分解酵素などが挙げられ、より具体的には、例えば、エンドグルカナーゼ、セロビオヒドロラーゼ、およびβ−グルコシダーゼ、アミラーゼ(例えば、グルコアミラーゼおよびα−アミラーゼ)、リパーゼなどが挙げられる。
細胞表層発現用ポリヌクレオチドは、さらにターミネーターを含むことができる。
アンカー(細胞表層局在タンパク質またはその細胞膜結合領域)をコードする配列を、分泌シグナル配列と共に所望の配置で、目的タンパク質をコードする配列(構造遺伝子)と連結し、この連結物をプロモーターの下流に配置する。細胞表層発現用ポリヌクレオチドは、例えば、目的タンパク質が発現されたときに、アンカー(細胞表層局在タンパク質またはその細胞膜結合領域)のN末端に連結されるように、上記配列を配置させる。すなわち、目的タンパク質をコードする配列は、アンカーをコードする配列の5’側に位置する。さらに、目的タンパク質をコードする配列は分泌シグナル配列の下流に配置される。
発現ベクターは、プラスミドベクターであってもよく、あるいは人工染色体であってもよい。酵母を宿主とする場合、ベクターの調製が容易であり、また酵母細胞の形質転換が容易である点で、プラスミドの形態が好ましい。DNAの取得の簡易化の点からは、酵母と大腸菌とのシャトルベクターであることが好ましい。必要に応じて、ベクターは、調節配列(オペレーター、エンハンサーなど)を含み得る。このようなベクターは、例えば、酵母の2μmプラスミドの複製開始点(Ori)とColE1の複製開始点とを有しており、酵母選択マーカー(以下に説明)および大腸菌の選択マーカー(薬剤耐性遺伝子など)を有する。
宿主として用いる酵母は、子のう菌酵母(Ascomycetous yeast)に属するものであればよく、特に限定されない。その中でもサッカロマイセス科(Saccharomycetaceae)に属するものが好ましく、サッカロマイセス属(Saccharomyces)であるものがより好ましい。
本発明によるセルロース分解酵素およびデンプン分解酵素からなる群から選択される少なくとも1つの酵素を細胞表層提示する酵母は、エタノールの製造に用いられ得る。1つの実施形態では、エンドグルカナーゼ、セロビオヒドロラーゼ、およびβ−グルコシダーゼからなる群から選択される少なくとも1つの酵素を細胞表層提示する酵母(本明細書中、「セルラーゼ細胞表層提示酵母」ともいう)である。このような酵母は、エンドグルカナーゼ、セロビオヒドロラーゼ、およびβ−グルコシダーゼからなる群から選択される2種の酵素;またはエンドグルカナーゼ、セロビオヒドロラーゼ、およびβ−グルコシダーゼを細胞表層提示する酵母であってもよい。別の実施形態では、α−アミラーゼおよび/またはグルコアミラーゼを細胞表層提示する酵母(本明細書中、「アミラーゼ細胞表層提示酵母」ともいう)である。酵母は、セルロース分解酵素およびデンプン分解酵素を共に表層提示するものであってよい。好ましくは、細胞表層発現用カセットとして、遺伝子Sed1のコーディング領域およびその遺伝子Sed1のプロモーターを含むカセットが用いられる。
サッカロマイセス・セレビシエ由来の細胞表層局在タンパク質遺伝子Sed1(以下、便宜上「SED1」ともいう)について、PCR法により、サッカロマイセス・セレビシエBY4741株ゲノムを鋳型とし、プライマー対(SED1a-XhoI-F(配列番号7)およびSED1a-BsrGI-R(配列番号8))を用いて増幅し、コーディング領域を含むDNA断片を調製した。この断片をXhoIおよびBsrGIで処理し、同様に処理したベクタープラスミドpIBG13(栄養要求性マーカー遺伝子HIS3、およびBGL1発現カセット(すなわち、GAPDH(グリセルアルデヒド−3’−リン酸デヒドロゲナーゼ)プロモーター、リゾプス・オリゼ由来グルコアミラーゼの分泌シグナルペプチド配列、BGL1のコーディング領域、α−アグルチニン遺伝子の3’側の半分の領域(α−アグルチニン遺伝子のコーディング領域の991位から1953位までのヌクレオチドからなる領域)、およびコーディング領域下流445bpのターミネーター領域がこの順に配置されているカセット)を有する表層発現用ベクター:非特許文献8)に連結した。得られたプラスミドをpIBG13Sと命名した。
BGL1、およびEGII遺伝子の各プラスミド(pIBG-PG-Agα1、またはpIEG-Agα1)をNdeIで処理し、それぞれ酵母サッカロマイセス・セレビシエBY4741株(MATα his3 leu2 met15 ura3株)に供し、酢酸リチウム法により形質転換した。これらの形質転換株を各遺伝子のGap−Agα1形質転換株と称する。
BGL1遺伝子のGap−Agα1形質転換株、Gap−Sed1形質転換株、Sed1−Agα1形質転換株、およびSed1−Sed1形質転換株について、β−グルコシダーゼ(BGL)活性を検討した。
(1)菌体を蒸留水で2回洗浄;
(2)反応液500μL(組成:10mM pNPG(p−ニトロフェニル−β−D−グルコピラノシド)100μL(最終濃度2mM);500mM クエン酸ナトリウム緩衝液(pH5.0) 50μL(最終濃度50mM);蒸留水250μL;および酵母100μL)(最終菌体濃度1〜10g湿潤菌体/L))を調製し、500rpm、30℃にて10分間反応;
(3)反応終了後、3M Na2CO3 500μLを加え反応を停止;そして
(4)10,000gで5分間遠心後、上清の400nmにおける吸光度ABS400を測定。1分間で1μmolのpNP(p−ニトロフェノール)を遊離する酵素量を1Uとする。
EGII遺伝子のGap−Agα1形質転換株、Gap−Sed1形質転換株、Sed1−Agα1形質転換株、およびSed1−Sed1形質転換株について、エンドグルカナーゼ(EG)活性を検討した。
(1)菌体を蒸留水で2回洗浄;
(2)反応液2500μL(組成:セラザイムCタブレット(Megazyme社製)1錠;500mM クエン酸ナトリウム緩衝液(pH5.0) 250μL(最終濃度50mM);蒸留水2000μL;および酵母250μL(最終菌体濃度10g湿潤菌体/L))を調製し、静置、38℃にて4時間反応;
(3)反応終了後、10,000gで5分間遠心後、上清の590nmの吸光度ABS590を測定。
BGL1遺伝子のSed1−Sed1形質転換株、Gap−Agα1形質転換株、およびCwp2−Cwp2形質転換株について、実施例3と同様にして、β−グルコシダーゼ(BGL)活性を検討した。
EGII−CBHII遺伝子共発現型Sed1−Sed1形質転換株、EGII−CBHII遺伝子共発現型Gap−Agα1形質転換株、および空ベクター導入株について、水熱処理稲ワラ加水分解に対する効果を検討した。
水熱処理稲ワラ 100g乾燥重/L
酵母エキス 10g/L
ペプトン 20g/L
クエン酸ナトリウム緩衝液 50mM(pH5.0)
酵母菌体 100g湿潤菌体重量/L
合計 10mL
上記を回転式発酵装置に入れ、市販酵素剤を添加せずに、38℃、35rpm、96時間反応させた。
BGL1遺伝子のSed1−Sed1形質転換株、Cwp2−Cwp2形質転換株、およびSed1−Cwp2形質転換株について、実施例3と同様にして、β−グルコシダーゼ(BGL)活性を検討した。
本実施例では、BY4741株およびそのSED1遺伝子破壊株(BY4741 SED1Δ株)のそれぞれについて、遺伝子Sed1のコーディング領域およびプロモーターを用いて調製したβ−グルコシダーゼ細胞表層提示酵母のβ−グルコシダーゼ(BGL)活性を検討した。
本実施例では、BY4741株およびそのSSD1遺伝子破壊株(BY4741 SSD1Δ株)のそれぞれについて、遺伝子Sed1のコーディング領域およびプロモーターを用いて調製したβ−グルコシダーゼ細胞表層提示酵母のβ−グルコシダーゼ(BGL)活性を検討した。
本実施例では、BY4741株についてSED1遺伝子およびSSD1遺伝子の両遺伝子を破壊した株(二重破壊株)を、BY4741株、BY4741 SED1Δ株およびBY4741 SSD1Δ株とともに、遺伝子Sed1のコーディング領域およびプロモーターを用いて調製したβ−グルコシダーゼ細胞表層提示酵母のβ−グルコシダーゼ(BGL)活性について検討した。
EGII遺伝子のSed1−Sed1形質転換株およびGap−Agα1形質転換株について、エタノール生産能を検討した。
水熱処理稲ワラ 100g乾燥重/L
酵母エキス 10g/L
ペプトン 20g/L
クエン酸ナトリウム緩衝液 50mM(pH5.0)
酵母菌体 100g湿潤菌体重量/L
酵素剤(Ctec2:Novozymes社製)1FPU/10mL
合計 10mL
リゾプス・オリゼ由来グルコアミラーゼ遺伝子、およびストレプトコッカス・ボビス由来α−アミラーゼ遺伝子の調製は、以下のように行った。
α−アミラーゼ遺伝子のプラスミド(pISpAA-Sed1)をNdeIで処理し、酵母サッカロマイセス・セレビシエBY4741株(MATα his3 leu2 met15 ura3株)に供し、酢酸リチウム法により形質転換した。この形質転換株をα−アミラーゼ遺伝子のSed1−Sed1形質転換株と称する。
α−アミラーゼ−グルコアミラーゼ遺伝子共発現型Sed1−Sed1形質転換株について、α−アミラーゼおよびグルコアミラーゼ活性を検討した。
α−アミラーゼ−グルコアミラーゼ遺伝子共発現型Sed1−Sed1形質転換株について、生デンプンからのエタノール生産能を検討した。
生デンプン 200g乾燥重/L
酵母エキス 10g/L
ペプトン 20g/L
酵母菌体 50g湿潤菌体重量/L
合計 5mL
上記を回転式発酵装置に入れ、市販酵素剤を添加せずに、30℃、35rpm、120時間反応させた。
Claims (8)
- 細胞表層発現用ポリヌクレオチドであって、該ポリヌクレオチドは、プロモーター、分泌シグナル配列、目的タンパク質をコードする配列、および細胞表層局在タンパク質またはその細胞膜結合領域をコードする配列を含み、該プロモーターが、該細胞表層局在タンパク質をコードする遺伝子のプロモーターである、ポリヌクレオチド。
- 前記細胞表層局在タンパク質がSED1またはCWP2である、請求項1に記載のポリヌクレオチド。
- 請求項1または2に記載の細胞表層発現用ポリヌクレオチドを含む、発現ベクター。
- 請求項1または2に記載の細胞表層発現用ポリヌクレオチドまたは請求項3に記載の発現ベクターが導入された、形質転換酵母。
- SED1およびSSD1からなる群より選択される少なくとも1つを欠損している宿主酵母から得られる、請求項4に記載の形質転換酵母。
- SED1およびSSD1を欠損している宿主酵母から得られる、請求項5に記載の形質転換酵母。
- セルロース分解酵素およびデンプン分解酵素からなる群から選択される少なくとも1つの酵素を細胞表層提示する、請求項4から6のいずれかに記載の形質転換酵母。
- エタノールを製造する方法であって、
請求項7に記載の形質転換酵母を発酵培養する工程を含む、方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2013062795 | 2013-03-25 | ||
JP2013062795 | 2013-03-25 | ||
PCT/JP2014/058189 WO2014157141A1 (ja) | 2013-03-25 | 2014-03-25 | 細胞表層発現用ポリヌクレオチド |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2014157141A1 true JPWO2014157141A1 (ja) | 2017-02-16 |
JP6335161B2 JP6335161B2 (ja) | 2018-05-30 |
Family
ID=51624112
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2015508514A Active JP6335161B2 (ja) | 2013-03-25 | 2014-03-25 | 細胞表層発現用ポリヌクレオチド |
Country Status (3)
Country | Link |
---|---|
US (1) | US9751917B2 (ja) |
JP (1) | JP6335161B2 (ja) |
WO (1) | WO2014157141A1 (ja) |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015033948A1 (ja) * | 2013-09-04 | 2015-03-12 | 関西化学機械製作株式会社 | エタノールの製造方法 |
JP6839424B2 (ja) * | 2016-12-27 | 2021-03-10 | 国立大学法人 鹿児島大学 | 真菌におけるタンパク質の選択的分泌技術 |
JP7194112B2 (ja) * | 2017-03-13 | 2022-12-21 | ラレマンド ハンガリー リクィディティー マネジメント エルエルシー | 細胞結合型異種タンパク質を発現する組換え酵母宿主細胞 |
JP6982438B2 (ja) * | 2017-09-06 | 2021-12-17 | 公立大学法人大阪 | デンプン分解酵素、それをコードする核酸、及びその利用 |
EP4010634A4 (en) | 2019-08-07 | 2023-09-13 | ANH Innovation, Inc. | MOBILE RECIRCULATION GRID WITH PLENUM AND DIFFUSER |
KR102268749B1 (ko) * | 2020-05-06 | 2021-06-23 | 전북대학교산학협력단 | 히스타민 결합 단백질 및 효모 표면 발현 단백질을 포함하는 재조합 효모 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005245335A (ja) * | 2004-03-04 | 2005-09-15 | Gekkeikan Sake Co Ltd | 目的タンパク質提示酵母及びその利用 |
JP2008086310A (ja) * | 2006-09-04 | 2008-04-17 | Gekkeikan Sake Co Ltd | セルロース分解酵素を表層提示する酵母及びその利用 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2007189909A (ja) | 2006-01-17 | 2007-08-02 | Gekkeikan Sake Co Ltd | 新規表層提示アンカー |
JP5780576B2 (ja) | 2009-07-08 | 2015-09-16 | 国立大学法人神戸大学 | セルロース分解性酵母およびその作製方法 |
JP2011160727A (ja) | 2010-02-10 | 2011-08-25 | Kobe Univ | 高温でセルロースからエタノールを生産する方法 |
JP2011167096A (ja) | 2010-02-17 | 2011-09-01 | Kobe Univ | バイオマスからのエタノールの生産方法 |
JP2012139211A (ja) | 2010-12-16 | 2012-07-26 | Kobe Univ | エタノールの生産方法 |
-
2014
- 2014-03-25 JP JP2015508514A patent/JP6335161B2/ja active Active
- 2014-03-25 US US14/778,250 patent/US9751917B2/en not_active Expired - Fee Related
- 2014-03-25 WO PCT/JP2014/058189 patent/WO2014157141A1/ja active Application Filing
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005245335A (ja) * | 2004-03-04 | 2005-09-15 | Gekkeikan Sake Co Ltd | 目的タンパク質提示酵母及びその利用 |
JP2008086310A (ja) * | 2006-09-04 | 2008-04-17 | Gekkeikan Sake Co Ltd | セルロース分解酵素を表層提示する酵母及びその利用 |
Non-Patent Citations (10)
Title |
---|
BIOTECHNOLOGY FOR BIOFUELS [ONLINE], vol. Vol.7, JPN6014026638, 14 January 2014 (2014-01-14), pages 8 (doi:10.1186/1754-6834-7-8) * |
BIOTECHNOLOGY LETTERS, vol. Vol.32, JPN6014026637, 2010, pages 1131 - 1136 * |
FEMS MICROBIOLOGY LETTERS, vol. Vol.162, JPN6014026631, 1998, pages 249 - 255 * |
INOKUMA, K. ET AL.: "Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and", BIOTECHNOLOGY FOR BIOFUELS [ONLINE], vol. Vol.7, JPN6014026638, 14 January 2014 (2014-01-14), pages 8 (doi:10.1186/1754-6834-7-8) * |
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, vol. 109, no. 5, JPN6014026633, 2010, pages 442 - 446 * |
KOTAKA, A. ET AL.: "Enhancement of β-glucosidase activity on the cell-surface of sake yeast by disruption of SED1", JOURNAL OF BIOSCIENCE AND BIOENGINEERING, vol. 109, no. 5, JPN6014026633, 2010, pages 442 - 446 * |
PROC. NATL. ACAD. SCI. USA, vol. 100, no. 5, JPN6014026635, 2003, pages 2766 - 2770 * |
RAM, A. F. J. ET AL.: "Green fluorescent protein-cell wall fusion proteins are covalently incorporated into the cell wall o", FEMS MICROBIOLOGY LETTERS, vol. Vol.162, JPN6014026631, 1998, pages 249 - 255, XP009076367, DOI: doi:10.1111/j.1574-6968.1998.tb13006.x * |
SU, G.-D. ET AL.: "Surface display of active lipase in Pichia pastoris using Sed1 as an anchor protein", BIOTECHNOLOGY LETTERS, vol. Vol.32, JPN6014026637, 2010, pages 1131 - 1136, XP019813452 * |
WHEELER, R. T. ET AL.: "A Saccharomyces cerevisiae mutant with increased virulence", PROC. NATL. ACAD. SCI. USA, vol. 100, no. 5, JPN6014026635, 2003, pages 2766 - 2770 * |
Also Published As
Publication number | Publication date |
---|---|
WO2014157141A1 (ja) | 2014-10-02 |
US20160326224A1 (en) | 2016-11-10 |
US9751917B2 (en) | 2017-09-05 |
JP6335161B2 (ja) | 2018-05-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11332728B2 (en) | Yeast strains for the expression and secretion of heterologous proteins at high temperatures | |
Manavalan et al. | Secretome analysis of Ganoderma lucidum cultivated in sugarcane bagasse | |
CA2657684C (en) | Construction of highly efficient cellulase compositions for enzymatic hydrolysis of cellulose | |
JP6335161B2 (ja) | 細胞表層発現用ポリヌクレオチド | |
CA3021166C (en) | Heterologous expression of fungal cellobiohydrolases in yeast | |
JP6537076B2 (ja) | 分泌シグナルペプチドならびにそれを利用したタンパク質の分泌および細胞表層提示 | |
DK2483415T3 (en) | RECOMBINANT C1 B-glucosidase FOR MANUFACTURE OF sugars from cellulosic BIOMASS | |
US20130280764A1 (en) | Method of improving the activity of cellulase enzyme mixtures in the saccharification (ligno)cellulosic material | |
WO2010148148A2 (en) | β-GLUCOSIDASE VARIANTS | |
US8906689B2 (en) | Endoglucanase variants | |
US20200347371A1 (en) | Variants of exoglucanases having improved actvity and uses thereof | |
WO2015033948A1 (ja) | エタノールの製造方法 | |
CA2780974C (en) | Mutli-cellulase enzyme compositions for hydrolysis of cellulosic biomass | |
WO2010101158A1 (ja) | クロストリジウム セルロボランス由来新規遺伝子及びその利用 | |
Mo et al. | Direct ethanol production from steam-exploded corn stover using a synthetic diploid cellulase-displaying yeast consortium | |
KR101106061B1 (ko) | 바이오 에탄올 생산능이 향상된 재조합 벡터 및 그 형질 전환체 | |
JP2021090384A (ja) | セルラーゼを細胞表層発現する形質転換酵母 | |
JP7319652B2 (ja) | タンパク質細胞表層発現酵母 | |
KR102092429B1 (ko) | 저온에서 강화된 베타-글루코시다아제 활성을 갖는 폴리펩티드 | |
CN111088244B (zh) | 蛋白酶基因在促进纤维素酶生产和复杂氮源利用中的应用 | |
JP2011160727A (ja) | 高温でセルロースからエタノールを生産する方法 | |
Sakwa | Cloning and expression of fungal alpha-amylase genes in Saccharomyces cerevisiae with integrated glucoamylase gene for raw starch conversion into bioethanol | |
Liao | Use of genetically modified saccharomyces cerevisiae to convert soluble starch directly to bioethanol |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20170215 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20171107 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20171129 |
|
RD04 | Notification of resignation of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7424 Effective date: 20180410 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20180424 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20180427 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6335161 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |