Enzymes for Starch Processing

A polypeptide having alpha-amylase activity, selected from the group consisting of:

• (a) a polypeptide having an amino acid sequence which has at least 75% homology with amino acids for the mature polypeptide selected from the group consisting of amino acids 1-441 in SEQ ID NO:
• 14, as amino acids 1-471 in SEQ ID NO:
• 18, as amino acids 1-450 in SEQ ID NO:
• 20, as amino acids 1-445 in SEQ ID NO:
• 22, as amino acids 1-498 in SEQ ID NO:
• 26, as amino acids 18-513 in SEQ ID NO:
• 28, as amino acids 1-507 in SEQ ID NO:
• 30, as amino acids 1-481 in SEQ ID NO:
• 32, as amino acids 1-495 in SEQ ID NO:
• 34, as amino acids 1-477 in SEQ ID NO:
• 38, as amino acids 1-449 in SEQ ID NO:
• 42, as amino acids 1-442 in SEQ ID NO:
• 115, as amino acids 1-441 in SEQ ID NO:
• 117, as amino acids 1-477 in SEQ ID NO:
• 125, as amino acids 1-446 in SEQ ID NO:
• 131, as amino acids 41-481 in SEQ ID NO:
• 157, as amino acids 22-626 in SEQ ID NO:
• 159, as amino acids 24-630 in SEQ ID NO:
• 161, as amino acids 27-602 in SEQ ID NO:
• 163, as amino acids 21-643 in SEQ ID NO:
• 165, as amino acids 29-566 in SEQ ID NO:
• 167, as amino acids 22-613 in SEQ ID NO:
• 169, as amino acids 21-463 in SEQ ID NO:
• 171, as amino acids 21-587 in SEQ ID NO:
• 173, as amino acids 30-773 in SEQ ID NO:
• 175, as amino acids 22-586 in SEQ ID NO:
• 177, as amino acids 20-582 in SEQ ID NO:
• 179.
• (b) a polypeptide which is encoded by a nucleotide sequence (i) which hybridizes under at least low stringency conditions with nucleotides 1-1326 in SEQ ID NO:
• 13, nucleotides 1-1413 in SEQ ID NO:
• 17, nucleotides 1-1350 in SEQ ID NO:
• 19, nucleotides 1-1338 in SEQ ID NO:
• 21, nucleotides 1-1494 in SEQ ID NO:
• 25, nucleotides 52-1539 in SEQ ID NO:
• 27, nucleotides 1-1521 in SEQ ID NO:
• 29, nucleotides 1-1443 in SEQ ID NO:
• 31, nucleotides 1-1485 in SEQ ID NO:
• 33, nucleotides 1-1431 in SEQ ID NO:
• 37, nucleotides 1-1347 in SEQ ID NO:
• 41, nucleotides 1-1326 in SEQ ID NO:
• 114, nucleotides 1-1323 in SEQ ID NO:
• 116, nucleotides 1-1431 in SEQ ID NO:
• 124, nucleotides 1-1338 in SEQ ID NO:
• 130, nucleotides 121-1443 in SEQ ID NO:
• 156, nucleotides 64-1878 in SEQ ID NO:
• 158, nucleotides 70-1890 in SEQ ID NO:
• 160, nucleotides 79-1806 in SEQ ID NO:
• 162, nucleotides 61-1929 in SEQ ID NO:
• 164, nucleotides 85-1701 in SEQ ID NO:
• 166, nucleotides 64-1842 in SEQ ID NO:
• 168, nucleotides 61-1389 in SEQ ID NO:
• 170, nucleotides 61-1764 in SEQ ID NO:
• 172, nucleotides 61-2322 in SEQ ID NO:
• 174, nucleotides 64-1761 in SEQ ID NO:
• 176, nucleotides 58-1749 in SEQ ID NO:
• 178, or (ii) which hybridizes under at least medium stringency conditions with the cDNA sequence contained in the polynucleotides shown as nucleotides 1-1326 in SEQ ID NO:
• 13, as nucleotides 1-1413 in SEQ ID NO:
• 17, as nucleotides 1-1350 in SEQ ID NO:
• 19, as nucleotides 1-1338 in SEQ ID NO:
• 21, as nucleotides 1-1494 in SEQ ID NO:
• 25, as nucleotides 52-1539 in SEQ ID NO:
• 27, as nucleotides 1-1521 in SEQ ID NO:
• 29, as nucleotides 1-1443 in SEQ ID NO:
• 31, as nucleotides 1-1485 in SEQ ID NO:
• 33, as nucleotides 1-1431 in SEQ ID NO:
• 37, as nucleotides 1-1347 in SEQ ID NO:
• 41, as nucleotides 1-1326 in SEQ ID NO:
• 114, as nucleotides 1-1323 in SEQ ID NO:
• 116, as nucleotides 1-1431 in SEQ ID NO:
• 124, as nucleotides 1-1338 in SEQ ID NO:
• 130, as nucleotides 121-1443 in SEQ ID NO:
• 156, as nucleotides 64-1878 in SEQ ID NO:
• 158, as nucleotides 70-1890 in SEQ ID NO:
• 160, as nucleotides 79-1806 in SEQ ID NO:
• 162, as nucleotides 61-1929 in SEQ ID NO:
• 164, as nucleotides 85-1701 in SEQ ID NO:
• 166, as nucleotides 64-1842 in SEQ ID NO:
• 168, as nucleotides 61-1389 in SEQ ID NO:
• 170, as nucleotides 61-1764 in SEQ ID NO:
• 172, as nucleotides 61-2322 in SEQ ID NO:
• 174, as nucleotides 64-1761 in SEQ ID NO:
• 176, as nucleotides 58-1749 in SEQ ID NO:
• 178, or (iii) a complementary strand of (i) or (ii);
• and
• (c) a variant comprising a conservative substitution, deletion, and/or insertion of one or more amino acids in an acid amino sequence selected from the group consisting of amino acids 1-441 in SEQ ID NO:
• 14, amino acids 1-471 in SEQ ID NO:
• 18, amino acids 1-450 in SEQ ID NO:
• 20, amino acids 1-445 in SEQ ID NO:
• 22, amino acids 1-498 in SEQ ID NO:
• 26, amino acids 18-513 in SEQ ID NO:
• 28, amino acids 1-507 in SEQ ID NO:
• 30, amino acids 1-481 in SEQ ID NO:
• 32, amino acids 1-495 in SEQ ID NO:
• 34, amino acids 1-477 in SEQ ID NO:
• 38, amino acids 1-449 in SEQ ID NO:
• 42, amino acids 1-442 in SEQ ID NO:
• 115, amino acids 1-441 in SEQ ID NO:
• 117, amino acids 1-477 in SEQ ID NO:
• 125, amino acids 1-446 in SEQ ID NO:
• 131, amino acids 41-481 in SEQ ID NO:
• 157, amino acids 22-626 in SEQ ID NO:
• 159, amino acids 24-630 in SEQ ID NO:
• 161, amino acids 27-602 in SEQ ID NO:
• 163, amino acids 21-643 in SEQ ID NO:
• 165, amino acids 29-566 in SEQ ID NO:
• 167, amino acids 22-613 in SEQ ID NO:
• 169, amino acids 21-463 in SEQ ID NO:
• 171, amino acids 21-587 in SEQ ID NO:
• 173, amino acids 30-773 in SEQ ID NO:
• 175, amino acids 22-586 in SEQ ID NO:
• 177 and amino acids 20-582 in SEQ ID NO:
• 179.

A polypeptide having carbohydrate-binding affinity, selected from the group consisting of:

• (a) a polypeptide comprising an amino acid sequence which has at least 60% homology with a sequence selected from the group consisting of amino acids 529-626 of SEQ ID NO:
• 159, amino acids 533-630 of SEQ ID NO:
• 161, amino acids 508-602 of SEQ ID NO:
• 163, amino acids 540-643 of SEQ ID NO:
• 165, amino acids 502-566 of SEQ ID NO:
• 167, amino acids 513-613 of SEQ ID NO:
• 169, 492-587 of SEQ ID NO:
• 173, amino acids 30-287 of SEQ ID NO:
• 175, amino acids 487-586 of SEQ ID NO:
• 177, and amino acids 482-582 of SEQ ID NO:
• 179;
• (b) a polypeptide which is encoded by a nucleotide sequence which hybridizes under low stringency conditions with a polynucleotide probe selected from the group consisting of the complementary strand of a sequence selected from the group consisting of nucleotides 1585-1878 in SEQ ID NO:
• 158, nucleotides 1597-1890 in SEQ ID NO:
• 160, nucleotides 1522-1806 in SEQ ID NO:
• 162, nucleotides 1618-1929 in SEQ ID NO:
• 164, nucleotides 1504-1701 in SEQ ID NO:
• 166, nucleotides 1537-1842 in SEQ ID NO:
• 168, nucleotides 1474-1764 in SEQ ID NO:
• 172, nucleotides 61-861 in SEQ ID NO:
• 174, nucleotides 1459-1761 in SEQ ID NO:
• 176, and nucleotides 1444-1749 in SEQ ID NO:
• 178;
• (c) a fragment of (a) or (b) that has carbohydrate binding affinity.

A process comprising:

• (a) contacting a starch with a polypeptide comprising a catalytic module having alpha-amylase activity and a carbohydrate-binding module;
• (b) incubating said starch with said polypeptide;
• (c) fermenting to produce a fermentation product,
• (d) optionally recovering the fermentation product,
• wherein an enzyme having glucoamylase activity is either absent or present in an amount of no more than or even less than 0.5 AGU/g DS, more preferably no more than or even less than 0.4 AGU/g DS, even more preferably no more than or even less than 0.3 AGU/g DS, and most preferably no more than or even less than 0.1 AGU, such as no more than or even less than 0.05 AGU/g DS of starch substrate and wherein step a, b, c, and/or d may be performed separately or simultaneously.

A process comprising:

• (a) contacting a starch substrate with a yeast cell transformed to express a polypeptide comprising a catalytic module having alpha-amylase activity and a carbohydrate-binding module;
• (b) holding said starch substrate with said yeast;
• (c) fermenting to produce ethanol
• (d) optionally recovering ethanol
• wherein steps a, b, and c are performed separately or simultaneously.