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Method For Producing Protein

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]

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Notices, Deadlines & Correspondence

Patent Information

Application #
Filing Date
12 October 2018
Publication Number
42/2018
Publication Type
INA
Invention Field
BIOTECHNOLOGY
Status
Email
patent@depenning.com
Parent Application
Patent Number
Legal Status
Grant Date
2023-05-19
Renewal Date

Applicants

TORAY INDUSTRIES, INC.
L-10 Nihonbashi-Muromachi 2-chome, Chuoku, Tokyo 103-8666, Japan.

Inventors

1. KOBAYASHI, Koji
c/o Basic Research Center, Toray Industrieso Inc., 10-1, Tebiro 6-chome, Kamakura-shi, Kanagawa 248-8555, Japan.
2. HIRAMATSU, Shingo
c/o Basic Research Center, Toray Industrieso Inc., 10-1, Tebiro 6-chome, Kamakura-shi, Kanagawa 248-8555, Japan.
3. YAMADA, Katsushige
c/o Basic Research Center, Toray Industrieso Inc., 10-1, Tebiro 6-chome, Kamakura-shi, Kanagawa 248-8555, Japan.

Specification

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.

Documents

Application Documents

# 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

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