US 2013 0122553A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2013/0122553 A1 Maertens et al. (43) Pub. Date: May 16, 2013 (54) METABOLICALLY ENGINEERED Publication Classification ORGANISMS FOR THE PRODUCTION OF ADDED VALUE BO-PRODUCTS (51) Int. Cl. CI2N 5/8 (2006.01) CI2N 15/70 (2006.01) (75) Inventors: Jo Maertens, Gent (BE); Joeri (52) U.S. Cl. Beauprez, Bredene (BE); Marjan De CPC ................ CI2N 15/81 (2013.01); C12N 15/70 Mey, Gent (BE) (2013.01) USPC .. 435/97; 435/252.3:435/254.2:435/254.11: (73) Assignee: UNIVERSITEIT GENT, Gent (BE) 435/419,435/325; 435/252.33: 435/254.21 (21) Appl. No.: 13/809,340 (57) ABSTRACT The present invention relates to genetically engineered organ (22) PCT Fled: Jul. 12, 2011 isms, especially microorganisms such as bacteria and yeasts, for the production of added value bio-products such as spe (86) PCT NO.: PCT/EP1 1/61891 cialty saccharide, activated saccharide, nucleoside, glyco S371 (c)(1), side, glycolipid or glycoprotein. More specifically, the (2), (4) Date: Jan. 9, 2013 present invention relates to host cells that are metabolically engineered so that they can produce said valuable specialty (30) Foreign Application Priority Data products in large quantities and at a high rate by bypassing classical technical problems that occur in biocatalytical or Jul. 12, 2010 (EP) .................................. 1O1693O4.2 fermentative production processes. 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GCAGGGGCTTCCTTCCTGCGTTTCGACGGCACGCCGGGTCCACCGACAAAGACGGCA CATCAGGTCGCTGGCGGCGGAAATCACCGCTGICACCGGAAGAACCCGCAGGA ACACACAACGAACGGCAAAACGCTTGGTGCGCCGAGCACAACCGGCAGGCA GCGCGACCCGCACAAAAAGCGGCGCGCTAAGCTGICCCGGAAATGGGAGCG CCAGCACCCGGCAGGGACCCGACACCGCGCGCCGACGCGCTCCGGGCAACGG IGCTCAGGCGGCTGAAAGGATGACTGACAACGGCTGGTTCGCCGCGCGTCCG TCAGGCACGGAAGACGCAATAAGATCTACTGCGAAAGCTTCCTCGGGAAGAACATC GCAAGCAGATTGAGAAAGAAGCGGTTGAGATTGAGCGAAGTTCTGAAAAACGCGTA AACACATTTAATAAAAAAAGGGCGGTCGCAAGATCGCCCTTTTTTACGTATGACAAAC ACAGAATTGCCTGATGCGCTACGCTTATCAGGCCTACGAGGAT Figure 12 CGTACACGCGTTTACTTTGCGGCAGATGAACAAACGCTGCTGAAAAATGGTAA TCAGACCAAGCCGAAACATGTGCCAGGCACGCCGTATTGGGTGATCACCAACA CCAACACCGGCCGTAAATGCAGCATGATCGAACACATCATGCAGTCGATGCAA TTCCCGGCGGAATTGATTGAGAAGGTTTGCGGAACTATCTAAAACGTTGCAGA CAAAGGACAAAGCAGIGAGGCGGAGCGCCGAAGICCATACTCA GAGAAAGGAACTTCGGAAAGGAACAAGGAGGAATCATAGACACATTT AATAAAAAAAGGGCGGTCGCAAGATCGCCCTTTTTTACGTATGACAAACACAG AATTGCCTGATGCGCTACGCTTATCAGGCCTACGAGGATGGTGCAATATATTG AATTTAAGCGATTTTGTAGGCCGGATAAGGCGTTCACGCCGCATCCGGCAAAA ACAACGAACACTTTGTCAACAAACTGAGTAGC Patent Application Publication May 16, 2013 Sheet 13 of 22 US 2013/O122553 A1 Figure 13 CGTACACGCGTTTACTTTGCGGCAGATGAACAAACGCTGCTGAAAAATGGTAATCA GACCAAGCCGAAACATGTGCCAGGCACGCCGTATTGGGTGAT CACCA ACACCA ACA CCGGCCGTAAATGCAGCATGATCGAACACATCATGCAGTCGATGCAATTCCCGGCG GAATTGATTGAGAAGGTTTGCGGAACTATCTAAAACGTTGCAGACAAAGGACAAAG CAGTGAGGCTGGAGCGCCGAAGCCAACTTCAGAGAAAGGAACTIC GGAAAGGAACAAGGAGGAATTCATAGGACCGAAICCCGGGCGGCCGCTCA TATAAATCGACAGGAAAGAGGAAAAGGACAATCCAG GCGGCCGCCTAGAAGAAGCTGGGATCCGTCGACCCGAATCGGAGGAAACAAA GATGGGGGGCTCATCATCATCATCATCATGGATGGCAGCATGAGAAAGGAG AAGAACCACGGAGGCCCAACGGAA AGAGGGAGAA GGGCACAAATCGTCAGTGGAGAGGGGAAGGGAGCAACAACGGAAAAC ACCCAAATTTATTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTG ICACTACTTTCGGGTATGGGTTCAATGCTTTGCNAGATACCCAGATCATAGAAA CAGCATGACTTTTCAAGAGGCCAGCCCGAAGGTAGACAGGAAAGAACTA ACAAAGATGACGGGAACTACAAGACACGGCGAAGTCAAGTTGAAGGG ATACCCTTGAAAGAATCGAGAAAAGGATTGAAAAGAAGATGGAAAC ATTCTTGGACACAAATGGAAACAACAAACCACACAATGAACACAGGC AGACAAACAAAAGAATGGAATCAAAGAACT CAAAAAGACACAACATTGAAG ATGGAAGCGTCAACTAGCAGACCAATCAACAAAAACTCCAAGGCGATGGC CCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATCTGCCCTTTCGAAAGA TCCCAACGAAAAGAGAGACCACATGGCCTTCGAGTGAACAGCGCGGGA TACACATGGCATGGATGAACTATACAAATAACGCAGGCGACCAAGGGAGAG CTCCCAACGCGGGAGCAGGCATGCAAGCTIGGCTGTTTGGCGGATGAGAGAA GACAGCCGAACAGAAAACAGAACGCAGAAGCGGTCGATAAAACAGA ATTTGCCTGGCGGCAGAGCGCGGGGTCCCACCGACCCCAGCCGAACTCAGAA GIGAAACGCCGAGCGCCGAGGAGGGGGGCTCCCCAIGCGAGAGTAGGGAA CGCCAGGCACAAAAAAACGAAAGGCCAGCGAAAGACGGGCCCG ACTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAATCCGCCGGGAGCGGA TTGAACGGCGAAGCAACGGCCCGGAGGGGGCGGGCAGGACGCCCGCCATAAA CTGCCAGGCATCAAATAAGCAGAAGGCCACCGACGGAGCTACTGCGAAAGCT TCCTCGGTGAAGAACATCGCAAGCAGATTGAGAAAGAAGCGGTTGAGATTGTTAGC GAAGTTCTGAAAAACGCGTAACTACTGCGAAAGCTTCCTCGGTGAAGAACATCGCA AGCAGATTGAGAAAGAAGCGGTTGAGATTGTTAGCGAAGTTCTGAAAAACGCGTAA ACACATTTAATAAAAAAAGGGCGGTCGCAAGATCGCCCTTTTTTACGTATGACAAA CACAGAATTGCCTGATGCGCTACGCTTATCAGGCCTACGAGGATGGTGCAATATAT TGAATTTAAGCGATTTTGTAGGCCGGATAAGGCGTTCACGCCGCATCCGGCAAAAA CAACGAACACTTTGTCAACAAACTGAGTAGCTCAAGGAAATCCCA Patent Application Publication May 16, 2013 Sheet 14 of 22 US 2013/O122553 A1 Figure 14 CGTACACGCGTTTACTTTGCGGCAGATGAACAAACGCTGCTGAAAAATGGTAATC AGACCAAGCCGAAACATGTGCCAGGCACGCCGTATTGGGTGATCACCAACACCAA CACCGGCCGTAAATGCAGCATGATCGAACACATCATGCAGTCGATGCAATTCCCG GCGGAATTGATTGAGAAGGTTTGCGGAACTATCTAAAACGTTGCAGACAAAGGAC AAAGCAACGAAAGGCTCAGTCGAAAGACTGGGCCCGTACTGGG TCGGTGAACGCCCCGAGTAGGACAAATCCGCCGGGAGCGGATGAACGG CGAAGCAACGGCCCGGAGGGTGGCGGGCAGGACGCCCGCCAAAACTGCCAGGCA TCAAATAAGCAGAAGGCCACCTGACGGAGGCCTTGCGGGCCAGGCCA AGCTTGCATGCAGAGCAGCATTACACGTCTTGAGCGATTGTGTAGGCGGAGC GCTTCGAAGTTCCTATACTTCAGAGAATAGGAACTTCGGAATAGGAACTTCA AGATCCCCTCACGCTGCCGCAAGCACTCAGGGCGCAAGGGCTGCTAAAGGAAGCG GAACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGC TACTGGGCTACTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTT GCAGGGGCACAGGCGATAGCTAGACTGGGCGGTTTAGGACAGCAAGCGA ACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTA AACGGAGGCT(TCGCCGCCAAGGACTGATGGCGCAGGGGATCAAGATCG ATCAAGAGACAGGATGAGGATCGTTTCGCAAGATCCCCTCACGCTGCCGCAAGCA CTCAGGGCGCAAGGGCGCTAAAGGAAGCGGAACACGTAGAAAGCCAGCCGCAG AAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGACAAGGGAAA ACGCAAGCGCAAAGAGAAAGCAGGTAGCTGCAGGGGCTTACAGGCGATAGCT AGACTGGGCGGTTTAGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCC TCTGGAAGGTTGGGAAGCCCTGCAAAGAAACGGATGGCCTTGCCGCCAA GGATCGATGGCGCAGGGGACAAGATCGATCAAGAGACAGGATGAGGATCGT CGCGCGGGACTCGGGGCGAAAGACCGACCAAGCGACGCCCAACCTGCCAT CACGAGATTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGGCTTCGGAATCGT TTTCCGGGACGCCGGCTGGAGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTC TCTACTGCGAAAGCTTCCTCGGTGAAGAACATCGCAAGCAGATTGAGAAAGAAGC GGTTGAGATTGTTAGCGAAGTTCTGAAAAACGCGTAAACACATTTAATAAAAAAA