| US 7,592,420 B2 | ||
| Expression in filamentous fungi of protease inhibitors and variants thereof | ||
| Hans De Nobel, Almere (Netherlands); David A. Estell, San Mateo, Calif. (US); Wei Liu, Palo Alto, Calif. (US); Scott D. Power, San Bruno, Calif. (US); Brian Schmidt, Half Moon Bay, Calif. (US); and Huaming Wang, Fremont, Calif. (US) | ||
| Assigned to Genencor International, Inc., Palo Alto, Calif. (US) | ||
| Filed on Aug. 30, 2007, as Appl. No. 11/848,196. | ||
| Application 11/848196 is a continuation of application No. 10/971596, filed on Oct. 22, 2004, granted, now 7,279,564. | ||
| Claims priority of provisional application 60/518154, filed on Nov. 06, 2003. | ||
| Prior Publication US 2008/0234465 A1, Sep. 25, 2008 | ||
| This patent is subject to a terminal disclaimer. | ||
| Int. Cl. C07K 14/42 (2006.01); C07K 14/81 (2006.01) | ||
| U.S. Cl. 530—324 [530/378; 435/69.2] | 14 Claims |
| 1. A composition comprising a protease inhibitor selected from the group consisting of:
a Bowman-Birk inhibitor (BBI) comprising SEQ ID NO:7 wherein residue 13 is Ala or is changed to Ser, Gly or Gln, and wherein
residues 15-21 and/or 42-48 have been replaced with a variant sequence; and
a Soybean Trypsin Inhibitor (STI) comprising SEQ ID NO:12 wherein residues 40-85 and/or 137-144 have been replaced with a
variant sequence;
wherein said variant sequence is selected from the group consisting of:
a VEGF binding peptide consisting of YNLYGWT (SEQ ID NO:77);
a complement protein c2 binding peptide consisting of GRKIPIQ (SEQ ID NO:78);
a complement protein c3 binding peptide consisting of ARSNLDE (SEQ ID NO:79);
a complement protein c4 binding peptide consisting of QRALPIL (SEQ ID NO:80);
a complement protein c5 binding peptide consisting of GRLHMKT (SEQ ID NO:81);
a complement factor B binding peptide consisting of KRKIVLD (SEQ ID NO:82);
a membrane metalloprotease 2 binding peptide consisting of AAMFGPA (SEQ ID NO:83);
a membrane metalloprotease 12 binding peptide consisting of GALGLFG (SEQ ID NO:84);
a cotton binding peptide consisting of EPLIHQR (SEQ ID NO:85); and
a cotton binding peptide consisting of SAFRGPT (SEQ ID NO:86),
wherein said protease inhibitor is produced in a filamentous fungal cell according to a method comprising,
a) introducing a DNA construct into a filamentous fungal cell, wherein said DNA construct comprises a promoter showing transcription
activity in the filamentous fungal cell and which is operably linked to a heterologous DNA sequence encoding the protease
inhibitor,
b) culturing the filamentous fungal cell under suitable culture conditions to allow expression of the heterologous DNA sequence,
and
c) producing the protease inhibitor.
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