| US 7,592,157 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. 11/848,213. | ||
| Application 11/848213 is a division 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 2008/0131932 A1, Jun. 05, 2008 | ||
| This patent is subject to a terminal disclaimer. | ||
| Int. Cl. C07K 14/42 (2006.01); C07K 14/81 (2006.01) | ||
| U.S. Cl. 435—69.2 [530/324; 530/378] | 21 Claims |
| 1. A method for producing a protease inhibitor in a filamentous fungal cell 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 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,
b) culturing the filamentous fungal cell under suitable culture conditions to allow expression of the heterologous DNA sequence,
and
c) producing the protease inhibitor;
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).
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