Transglutaminase variants

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.

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 .