Transglutaminase variants

Patent metadata

Field	Value
Publication number	US-2022220456-A1
Application number	US-202217705772-A
Country	US
Kind code	A1
Filing date	Mar 28, 2022
Priority date	Nov 7, 2017
Publication date	Jul 14, 2022
Grant date	—

Abstract

The present invention provides engineered transglutaminase enzymes, polynucleotides encoding the enzymes, compositions comprising the enzymes, methods of producing these enzymes, and methods of using the engineered transglutaminase enzymes.

First claim

We claim:

1 . An engineered polynucleotide encoding an engineered transglutaminase, wherein said engineered transglutaminase comprises a polypeptide sequence comprising at least 85% or more sequence identity to SEQ ID NO: 2, 6, 34, and/or 256.

2 . The engineered polynucleotide of claim 1 , wherein said engineered polynucleotide encodes an engineered transglutaminase comprising a polypeptide sequence having at least 85% or more sequence identity to SEQ ID NO:6, and at least one substitution or substitution set at one or more positions selected from positions 79, 101, 101/201/212/287, 101/201/285, 101/287, and 327, wherein said positions are numbered with reference to SEQ ID NO:6.

3 . The engineered polynucleotide of claim 1 , wherein said engineered polynucleotide encodes an engineered transglutaminase comprising a polypeptide sequence having at least 85% or more sequence identity to SEQ ID NO:2, and at least one substitution or substitution set at one or more positions selected from positions 48, 48/67/70, 48/67/70/181/203/256, 48/67/70/181/256/345, 48/67/70/181/296/345/373, 48/67/70/203/256/296/345, 48/67/70/203/256/345/354/373, 48/67/70/203/345, 48/67/70/256, 48/67/70/256/296/345/373, 48/67/203/256/296/373, 48/67/203/256/345, 48/70/170/203, 48/70/203/254/296/343, 48/70/203/256/345/373, 48/70/203/256/345, 48/70/203/373, 48/170/203, 48/170/203/254/296/346, 48/170/203/254/296/346/373, 48/170/203/254/346/373, 48/170/203/254/346, 48/170/203/296/343/346, 48/170/203/296/346/373, 48/170/203/343/346, 48/170/203/346, 48/170/203/346/373, 48/170/203/373, 48/170/254, 48/170/296, 48/170/296/343/346, 48/170/343/346, 48/181, 48/181/203/256/345, 48/181/203/345, 48/181/256/296/345, 48/181/296, 48/181/296/345, 48/203, 48/203/254/296, 48/203/254/296/343/373, 48/203/254/296/346/373, 48/203/254/346, 48/203/254/346/373, 48/203/256, 48/203/256/296/345, 48/203/296/343/346/373, 48/203/296/343/373, 48/203/296/346, 48/203/296/346/373, 48/203/343/346, 48/203/343/346/373, 48/203/345, 48/203/346, 48/203/346/373, 48/254/296, 48/254/346, 48/256, 48/256/296, 48/256/296/345, 48/296/345, 48/296/373, 48/343/346, 48/345/373, 67/256, 67/296/345, 68/74/190/215/346, 68/136/215/255/282/297/346, 68/136/215/297/346, 68/136/234, 68/158/174/234/282/297/346, 68/158/215/297/346, 68/215/297/346, 68/234, 68/282/297/346, 68/297/346, 74/136/174/282/346, 74/136/174/297/346, 74/136/346, 74/158/255/297, 74/255/346, 74/346, 136/158/190/215/255/297/346, 136/158/215/297/346, 136/174/215/255/282/297/346, 136/190/215/297/346, 136/215/234/282/297, 136/215/234/297/346, 136/215/297, 136/297/346, 158/215/255/346, 158/215/346, 170/203/254/296/343/346, 170/203/254/343/373, 170/203/343/346, 174/190/234/297/346, 174/215/234/297/346, 174/215/255/297/346, 174/282/297/346, 190/255/282/346, 190/297/346, 203/296, 203/343, 203/343/346, 203/346, 215/255/297/346, 215/234/297/346, 215/255/297/346, 215/297, 215/297/346, 215/346, 234/255/346, 255/297/346, 255/346, 297/346, 343/346/373, and 346, wherein said positions are numbered with reference to SEQ ID NO:2.

4 . The engineered polynucleotide of claim 1 , wherein said engineered polynucleotide encodes an engineered transglutaminase comprising a polypeptide sequence having at least 90% sequence identity to SEQ ID NO:2, 6, 34, and/or 256.

5 . The engineered polynucleotide of claim 1 , wherein said engineered polynucleotide encodes an engineered transglutaminase comprising a polypeptide sequence having at least 95% sequence identity to SEQ ID NO:2, 6, 34, and/or 256.

6 . The engineered polynucleotide of claim 1 , wherein said engineered polynucleotide encodes an engineered transglutaminase comprising a polypeptide sequence set forth in SEQ ID NO:2, 6, 34, or 256.

7 . The engineered polynucleotide of claim 1 , wherein said engineered polynucleotide encodes an engineered transglutaminase comprising a polypeptide sequence encoding a variant provided in Table 8.1, 9.1, 9.2, 10.1, and/or 11.1.

8 . The engineered polynucleotide of claim 1 , wherein said engineered polynucleotide encodes an engineered transglutaminase comprising a polypeptide sequence selected from the even-numbered sequences set forth in SEQ ID NOS: 4 to 756.

9 . An engineered polynucleotide comprising a polynucleotide sequence that is at least 85% or more identical to a sequence selected from the odd-numbered sequences set forth in SEQ ID NOS: 3 to 755.

10 . A vector comprising the engineered polynucleotide sequence of claim 1 .

11 . A vector comprising the engineered polynucleotide sequence of claim 9 .

12 . The vector of claim 10 , further comprising at least one control sequence.

13 . The vector of claim 11 , further comprising at least one control sequence.

14 . A host cell comprising the vector of claim 10 .

15 . A host cell comprising the vector of claim 11 .

Assignees

• Codexis Inc

Inventors

• Nazor Jovana
• Yang Jie
• Banerjee Goutami
• Zhang Xiyun
• Riggins James Nicholas
• Milczek Erika M
• Moore Jeffrey C

Classifications

• C07K14/62

  Insulins · CPC title

• C12N9/1044 Primary

  Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII · CPC title

• C12Y203/02013

  Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII · CPC title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title