Abstract A purified thermostable enzyme is derived from the archael bacterium Thermococcus GU5L5. The enzyme has a molecular weight of about 68.5 kilodaltons and has cellulase activity. The enzyme can be produced from native or recombinant host cells and can be used for the removal of arginine, phenylalanine, or methionine amino acids from the N-terminal end of peptides in peptide or peptidomimetic synthesis. The enzyme is selective for the L, or 'natural' enantiomer of the amino acid derivatives and is therefore useful for the production of optically active compounds. These reactions can be performed in the presence of the chemically more reactive ester functionality, a step which is very difficult to achieve with nonenzymatic methods.
An isolated polynucleotide `encoding an enzyme with amidase activity and which is` selected from the group consisting of: fragments of SEQ ID NO:1 that are at least 35 bases in length and that hybridize to a nucleic acid sequence encoding the polypeptide set forth in SEQ ID NO:2 under conditions that include 0.9 M NaCl, 5.0 mM NaH 2 PO 4 , 5.0 mM Na 2 EDTA, 0.5% SDS and 10× Denhardt's at about 45° C.
Claim 2
Dependent Claim
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An isolated polynucleotide `encoding an enzyme with amidase activity and which is` selected from the group consisting of: fragments of SEQ ID NO:1 that are at least 35 bases in length and that hybridize to a nucleic acid sequence encoding the polypeptide set forth in SEQ ID NO:2 under conditions that include 0.9 M NaCl, 5.0 mM NaH 2 PO 4 , 5.0 mM Na 2 EDTA, 0.5% SDS and 10× Denhardt's at about 45° C.
Claim 3
Dependent Claim
10 words
A polynucleotide of claim 1 wherein the polynucleotide is DNA.
Claim 4
Dependent Claim
10 words
A polynucleotide of claim 1 wherein the polynucleotide is RNA.
Claim 5
Dependent Claim
8 words
A vector comprising the polvnucleotide of claim 1.