Abstract: Disclosed is a method of producing a protein using a filamentous fungus, in which decrease in dissolved oxygen saturation during culture of the filamentous fungus can be suppressed even when the culture is scaled up. The method of producing a protein includes culturing a fungus belonging to the genus Trichoderma whose BXL1 gene was disrupted, using a biomass containing cellulose and xylan as an inducer. The use of the BXL1 gene-disrupted fungus belonging to the genus Trichoderma enables suppression of the decrease in dissolved oxygen saturation even when xylose and cellulose are used as inducers. [Figure 2]
WE CLAIM:
1. A method of producing a protein using a fungus belonging to the genus Trichoderma, said method comprising culturing a fungus belonging to the genus Trichoderma whose BXL1 gene was disrupted, using a biomass containing cellulose and xylan as an inducer.
2. The method according to claim 1, wherein the culturing is carried out adding said inducer to the culture medium to a concentration of at least 5% by weight (wt/wt).
3. The method according to claim 1 or 2, wherein said inducer has a xylan content of 15 to 40% by weight.
4. The method according to any one of claims 1 to 3, wherein said fungus belonging to the genus Trichoderma is Trichoderma reesei.
5. The method according to claim 4, wherein said Trichoderma reesei is a strain in which carbon catabolite repression is removed.
6. The method according to any one of claims 1 to 5, wherein said protein is a cellulase composition.
7. The method according to claim 6, wherein p-xylosidase specific activity in said cellulase composition in terms of an enzyme activity for degrading p-nitrophenyl-p-D-xylopyranoside per 1 mg of protein in said cellulase composition is 0.006 U/mg protein or less; wherein cellobiohydrolase specific activity in terms of an enzyme activity for degrading/>-nitrophenyl-p-D-lactopyranoside per 1 mg of protein in said cellulase is at least 0.1 U/mg protein; and wherein (3-glucosidase specific activity in terms of an enzyme activity for degrading /?-nitrophenyl-p-D-glucopyranoside per 1 mg of protein in said cellulase is at least 0.25 U/mg protein.
8. The method according to claim 7, wherein said p-xylosidase specific activity is 0.002 U/mg protein or less; and wherein said p-glucosidase specific activity is at least 0.3 U/mg protein.
9. A method of producing xylo-oligosaccharides, said method comprising hydrolyzing a biomass containing xylan and cellulose with said cellulase composition produced by said method according to any one of claims 6 to 8.
10. A method of producing xylo-oligosaccharides and glucose, said method comprising hydrolyzing a biomass containing xylan and cellulose with said cellulase composition produced by said method according to any one of claims 6 to 8.
11. A method of suppressing decrease in dissolved oxygen saturation when culturing a fungus belonging to the genus Trichoderma using a biomass containing cellulose and xylan as an inducer, said method comprising using a fungus belonging to the genus Trichoderma whose BXL1 gene was disrupted as said fungus belonging to the genus Trichoderma.
12. A cellulase composition having the following characteristics (a) to (d):
(a) (3-xylosidase specific activity in said cellulase composition in terms of an enzyme activity for degrading ^>-nitrophenyl-(3-D-xylopyranoside per 1 mg of protein in said cellulase composition is 0.006 U/mg protein or less;
(b) cellobiohydrolase specific activity in terms of an enzyme activity for degrading/>-nitrophenyl-(3-D-lactopyranoside per 1 mg of protein in said cellulase composition is at least 0.1 U/mg protein;
(c) (3-glucosidase specific activity in terms of an enzyme activity for degrading p-nitrophenyl-P-D-glucopyranoside per 1 mg of protein in said cellulase composition is at least 0.25 U/mg protein; and
(d) the protein concentration of said cellulase composition is 3 g/L or more.
13. The cellulase composition according to claim 12, wherein said protein concentration is 9 g/L or more.
14. The cellulase composition according to claim 12 or 13, whereinp-nitrophenyl-p-D-xylobioside-degrading activity per 1 mg of protein in said cellulase composition is at least 5 U/mg protein.
| # | Name | Date |
|---|---|---|
| 1 | 201847038786.pdf | 2018-10-12 |
| 2 | 201847038786-TRANSLATIOIN OF PRIOIRTY DOCUMENTS ETC. [12-10-2018(online)].pdf | 2018-10-12 |
| 3 | 201847038786-STATEMENT OF UNDERTAKING (FORM 3) [12-10-2018(online)].pdf | 2018-10-12 |
| 4 | 201847038786-SEQUENCE LISTING(PDF) [12-10-2018(online)].txt | 2018-10-12 |
| 5 | 201847038786-SEQUENCE LISTING [12-10-2018(online)].pdf | 2018-10-12 |
| 6 | 201847038786-PROOF OF RIGHT [12-10-2018(online)].pdf | 2018-10-12 |
| 7 | 201847038786-PRIORITY DOCUMENTS [12-10-2018(online)].pdf | 2018-10-12 |
| 8 | 201847038786-POWER OF AUTHORITY [12-10-2018(online)].pdf | 2018-10-12 |
| 9 | 201847038786-FORM 1 [12-10-2018(online)].pdf | 2018-10-12 |
| 10 | 201847038786-DRAWINGS [12-10-2018(online)].pdf | 2018-10-12 |
| 11 | 201847038786-DECLARATION OF INVENTORSHIP (FORM 5) [12-10-2018(online)].pdf | 2018-10-12 |
| 12 | 201847038786-COMPLETE SPECIFICATION [12-10-2018(online)].pdf | 2018-10-12 |
| 13 | 201847038786-CLAIMS UNDER RULE 1 (PROVISIO) OF RULE 20 [12-10-2018(online)].pdf | 2018-10-12 |
| 14 | abstract_201847038786.jpg | 2018-10-15 |
| 15 | Correspondence by Agent_Form1_17-10-2018.pdf | 2018-10-17 |
| 16 | 201847038786-FORM 3 [25-03-2019(online)].pdf | 2019-03-25 |
| 17 | 201847038786-FORM 18 [03-02-2020(online)].pdf | 2020-02-03 |
| 18 | 201847038786-FER.pdf | 2021-10-17 |
| 19 | 201847038786-OTHERS [28-02-2022(online)].pdf | 2022-02-28 |
| 20 | 201847038786-OTHERS [28-02-2022(online)]-1.pdf | 2022-02-28 |
| 21 | 201847038786-FORM-26 [28-02-2022(online)].pdf | 2022-02-28 |
| 22 | 201847038786-FORM-26 [28-02-2022(online)]-1.pdf | 2022-02-28 |
| 23 | 201847038786-FORM 3 [28-02-2022(online)].pdf | 2022-02-28 |
| 24 | 201847038786-FORM 3 [28-02-2022(online)]-1.pdf | 2022-02-28 |
| 25 | 201847038786-FER_SER_REPLY [28-02-2022(online)].pdf | 2022-02-28 |
| 26 | 201847038786-FER_SER_REPLY [28-02-2022(online)]-1.pdf | 2022-02-28 |
| 27 | 201847038786-DRAWING [28-02-2022(online)].pdf | 2022-02-28 |
| 28 | 201847038786-DRAWING [28-02-2022(online)]-1.pdf | 2022-02-28 |
| 29 | 201847038786-COMPLETE SPECIFICATION [28-02-2022(online)].pdf | 2022-02-28 |
| 30 | 201847038786-COMPLETE SPECIFICATION [28-02-2022(online)]-1.pdf | 2022-02-28 |
| 31 | 201847038786-CLAIMS [28-02-2022(online)].pdf | 2022-02-28 |
| 32 | 201847038786-CLAIMS [28-02-2022(online)]-1.pdf | 2022-02-28 |
| 33 | 201847038786-ABSTRACT [28-02-2022(online)].pdf | 2022-02-28 |
| 34 | 201847038786-ABSTRACT [28-02-2022(online)]-1.pdf | 2022-02-28 |
| 35 | 201847038786-US(14)-HearingNotice-(HearingDate-04-05-2022).pdf | 2022-04-06 |
| 36 | 201847038786-REQUEST FOR ADJOURNMENT OF HEARING UNDER RULE 129A [28-04-2022(online)].pdf | 2022-04-28 |
| 37 | 201847038786-US(14)-ExtendedHearingNotice-(HearingDate-02-06-2022).pdf | 2022-05-02 |
| 38 | 201847038786-Correspondence to notify the Controller [30-05-2022(online)].pdf | 2022-05-30 |
| 39 | 201847038786-Written submissions and relevant documents [17-06-2022(online)].pdf | 2022-06-17 |
| 40 | 201847038786-Retyped Pages under Rule 14(1) [17-06-2022(online)].pdf | 2022-06-17 |
| 41 | 201847038786-Annexure [17-06-2022(online)].pdf | 2022-06-17 |
| 42 | 201847038786-2. Marked Copy under Rule 14(2) [17-06-2022(online)].pdf | 2022-06-17 |
| 43 | 201847038786-US(14)-HearingNotice-(HearingDate-02-05-2023).pdf | 2023-04-13 |
| 44 | 201847038786-Correspondence to notify the Controller [28-04-2023(online)].pdf | 2023-04-28 |
| 45 | 201847038786-Written submissions and relevant documents [17-05-2023(online)].pdf | 2023-05-17 |
| 46 | 201847038786-Retyped Pages under Rule 14(1) [17-05-2023(online)].pdf | 2023-05-17 |
| 47 | 201847038786-2. Marked Copy under Rule 14(2) [17-05-2023(online)].pdf | 2023-05-17 |
| 48 | 201847038786-PatentCertificate19-05-2023.pdf | 2023-05-19 |
| 49 | 201847038786-IntimationOfGrant19-05-2023.pdf | 2023-05-19 |
| 1 | SearchreportE_03-08-2021.pdf |