Engineered phosphoglucose isomerase proteins with a protease cleavage site

• US 9,688,977 B2
• Filed: 08/05/2014
• Issued: 06/27/2017
• Est. Priority Date: 08/05/2013
• Status: Active Grant

• Current Assignee: GreenLight Biosciences, Inc.
• Original Assignee: GreenLight Biosciences, Inc.
• Inventors: Cunningham, Drew S., Blake, William Jeremy
• Primary Examiner(s): Swope, Sheridan
• Application Number:
  US14/451,708
• Publication Number:
  US 20150037868A1
• Time in Patent Office: 1,057 Days
• Field of Search: None
• CPC Class Codes: C07K 2319/50 containing protease site
• CPC Class Codes: C12N 9/1007 Methyltransferases (general) (2.1.1.)
• CPC Class Codes: C12N 9/1022 transferring aldehyde or ketonic groups (2.2)
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12N 9/92 Glucose isomerase (5.3.1.5; 5.3.1.9; 5.3.1.18)
• CPC Class Codes: C12Y 202/01001 Transketolase (2.2.1.1)
• CPC Class Codes: C12Y 203/01008 Phosphate acetyltransferase (2.3.1.8)
• CPC Class Codes: C12Y 503/01009 Glucose-6-phosphate isomerase (5.3.1.9)

Abstract

Various aspects and embodiments herein relate to recombinant proteins with at least one protease recognition sequence that can be inactivated by a cognate protease and methods of preparing such proteins. In some embodiments, recombinant phosphoglucose isomerase (Pgi) proteins are provided. In other embodiments, recombinant phosphotransacetylase (Pta) proteins are provided. In yet other embodiments, recombinant transketolase A (TktA) proteins are provided.

Claims

1. A recombinant protein comprising a polypeptide derived from SEQ ID NO:

17, said polypeptide comprising a human rhinovirus 3C protease recognition sequence located between amino acids 526 and 527 of SEQ ID NO;

17 wherein, said polypeptide has at least 95% identity with SEQ ID NO;

25 and has phosphoglucose isomerase activity.

2. The recombinant protein of claim 1, wherein the protease recognition sequence comprises the amino acid sequence of SEQ ID NO:

38.

3. The recombinant protein of claim 1, wherein the recombinant protein comprises the amino acid sequence of SEQ ID NO:

25.

4. The recombinant protein of claim 1, wherein the protease recognition sequence comprises the amino acid sequence of SEQ ID NO:

39, SEQ ID NO;

40, SEQ ID NO;

41, SEQ ID NO;

42, SEQ ID NO;

43, SEQ ID NO;

44, or SEQ ID NO;

45.