LISTA DE SECUENCIAS <110> CENTRO DE INGENIERIA GENETICA Y BIOTECNOLOGIA <120> LEVANASACARASA MODIFICADA PARA LA PRODUCCION OPTIMIZADA DE FRUCTOOLIGOSACARIDOS <130> LsdAMUT <140> <141> <150> 2021-0020 <151> 2021-03-30 <160> 34 <170> PatentIn Ver. 2.1 <210> 1 <211> 25 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador de la PCR <400> 1 ggatcgtttc ccatggcgca tgtac 25 <210> 2 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 2 gatgcgggca tgcacgtgnn ngtcgtcgaa acc 33 <210> 3 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 3 gatgcgggca tgcacgctnn ngtcgtcgaa acc 33 <210> 4 <211> 24 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador de la PCR <400> 4 ctggggcgga tccacgccga cttc 24 <210> 5 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 5 gcgcagaacg aaaacttcaa tttccgcgat ccg 33 <210> 6 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 6 cggatcgcgg aaattgaaga attcgttctg cgc 33 <210> 7 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 7 gcgcagaacg aatacttcaa tttccgcgat ccg 33 <210> 8 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 8 cggatcgcgg aaattgaagt attcgttctg cgc 33 <210> 9 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 9 gcgcagaacg aatggttcaa tttccgcgat ccg 33 <210> 10 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 10 cggatcgcgg aaattgaacc attcgttctg cgc 33 <210> 11 <211> 24 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador de la PCR <400> 11 ggcgccaggg tacccccgcg acgg 24 <210> 12 <211> 27 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 12 cgcagaacga attcttcnnn ttccgcg 27 <210> 13 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 13 ggcaataccg cgggccagca tggcgtcgcc aac 33 <210> 14 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 14 gttggcgacg ccatgctggc ccgcggtatt gcc 33 <210> 15 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 15 ggcaataccg cgggccagaa gggcgtcgcc aac 33 <210> 16 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 16 gttggcgacg cccttctggc ccgcggtatt gcc 33 <210> 17 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 17 ggcaataccg cgggccagtc gggcgtcgcc aac 33 <210> 18 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 18 gttggcgacg cccgactggc ccgcggtatt gcc 33 <210> 19 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 19 ggcaataccg cgggccaggc cggcgtcgcc aac 33 <210> 20 <211> 33 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 20 gttggcgacg ccggcctggc ccgcggtatt gcc 33 <210> 21 <211> 27 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 21 cgcagaacga attcttccac ttccgcg 27 <210> 22 <211> 27 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 22 cgcagaacga attcttccgc ttccgcg 27 <210> 23 <211> 27 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador en la mutagenesis del gen lsdA <400> 23 cgcagaacga attcttcaag ttccgcg 27 <210> 24 <211> 27 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador de la PCR <400> 24 agtgcgctat cgatagggaa tttcagc 27 <210> 25 <211> 27 <212> ADN <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Oligonucleótido empleado como cebador de la PCR <400> 25 ttactggtct agaaattggc gaacctg 27 <210> 26 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada H142E <400> 26 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Arg Glu Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Phe Phe Asn Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Arg 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 <210> 27 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada R141K-H142S <400> 27 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Lys Ser Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Phe Phe Asn Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Arg 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 <210> 28 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada F274A <400> 28 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Arg His Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Ala Phe Asn Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Arg 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 <210> 29 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada N276H <400> 29 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Arg His Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Phe Phe His Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Arg 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 <210> 30 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada N276R <400> 30 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Arg His Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Phe Phe Arg Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Arg 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 <210> 31 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada R304K <400> 31 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Arg His Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Phe Phe Asn Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Lys 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 <210> 32 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada N276H-R304K <400> 32 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Arg His Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Phe Phe His Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Lys 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 <210> 33 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada N276R-R304K <400> 33 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Arg His Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Phe Phe Arg Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Lys 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 <210> 34 <211> 554 <212> PRT <213> Secuencia Artificial <220> <223> Descripción de la Secuencia Artificial:Levanasacarasa modificada N276K-R304K <400> 34 Gln Gly Asn Phe Ser Arg Gln Glu Ala Ala Arg Met Ala His Arg Pro 1 5 10 15 Gly Val Met Pro Arg Gly Gly Pro Leu Phe Pro Gly Arg Ser Leu Ala 20 25 30 Gly Val Pro Gly Phe Pro Leu Pro Ser Ile His Thr Gln Gln Ala Tyr 35 40 45 Asp Pro Gln Ser Asp Phe Thr Ala Arg Trp Thr Arg Ala Asp Ala Leu 50 55 60 Gln Ile Lys Ala His Ser Asp Ala Thr Val Ala Ala Gly Gln Asn Ser 65 70 75 80 Leu Pro Ala Gln Leu Thr Met Pro Asn Ile Pro Ala Asp Phe Pro Val 85 90 95 Ile Asn Pro Asp Val Trp Val Trp Asp Thr Trp Thr Leu Ile Asp Lys 100 105 110 His Ala Asp Gln Phe Ser Tyr Asn Gly Trp Glu Val Ile Phe Cys Leu 115 120 125 Thr Ala Asp Pro Asn Ala Gly Tyr Gly Phe Asp Asp Arg His Val His 130 135 140 Ala Arg Ile Gly Phe Phe Tyr Arg Arg Ala Gly Ile Pro Ala Ser Arg 145 150 155 160 Arg Pro Val Asn Gly Gly Trp Thr Tyr Gly Gly His Leu Phe Pro Asp 165 170 175 Gly Ala Ser Ala Gln Val Tyr Ala Gly Gln Thr Tyr Thr Asn Gln Ala 180 185 190 Glu Trp Ser Gly Ser Ser Arg Leu Met Gln Ile His Gly Asn Thr Val 195 200 205 Ser Val Phe Tyr Thr Asp Val Ala Phe Asn Arg Asp Ala Asn Ala Asn 210 215 220 Asn Ile Thr Pro Pro Gln Ala Ile Ile Thr Gln Thr Leu Gly Arg Ile 225 230 235 240 His Ala Asp Phe Asn His Val Trp Phe Thr Gly Phe Thr Ala His Thr 245 250 255 Pro Leu Leu Gln Pro Asp Gly Val Leu Tyr Gln Asn Gly Ala Gln Asn 260 265 270 Glu Phe Phe Lys Phe Arg Asp Pro Phe Thr Phe Glu Asp Pro Lys His 275 280 285 Pro Gly Val Asn Tyr Met Val Phe Glu Gly Asn Thr Ala Gly Gln Lys 290 295 300 Gly Val Ala Asn Cys Thr Glu Ala Asp Leu Gly Phe Arg Pro Asn Asp 305 310 315 320 Pro Asn Ala Glu Thr Leu Gln Glu Val Leu Asp Ser Gly Ala Tyr Tyr 325 330 335 Gln Lys Ala Asn Ile Gly Leu Ala Ile Ala Thr Asp Ser Thr Leu Ser 340 345 350 Lys Trp Lys Phe Leu Ser Pro Leu Ile Ser Ala Asn Cys Val Asn Asp 355 360 365 Gln Thr Glu Arg Pro Gln Val Tyr Leu His Asn Gly Lys Tyr Tyr Ile 370 375 380 Phe Thr Ile Ser His Arg Thr Thr Phe Ala Ala Gly Val Asp Gly Pro 385 390 395 400 Asp Gly Val Tyr Gly Phe Val Gly Asp Gly Ile Arg Ser Asp Phe Gln 405 410 415 Pro Met Asn Tyr Gly Ser Gly Leu Thr Met Gly Asn Pro Thr Asp Leu 420 425 430 Asn Thr Ala Ala Gly Thr Asp Phe Asp Pro Ser Pro Asp Gln Asn Pro 435 440 445 Arg Ala Phe Gln Ser Tyr Ser His Tyr Val Met Pro Gly Gly Leu Val 450 455 460 Glu Ser Phe Ile Asp Thr Val Glu Asn Arg Arg Gly Gly Thr Leu Ala 465 470 475 480 Pro Thr Val Arg Val Arg Ile Ala Gln Asn Ala Ser Ala Val Asp Leu 485 490 495 Arg Tyr Gly Asn Gly Gly Leu Gly Gly Tyr Gly Asp Ile Pro Ala Asn 500 505 510 Arg Ala Asp Val Asn Ile Ala Gly Phe Ile Gln Asp Leu Phe Gly Gln 515 520 525 Pro Thr Ser Gly Leu Ala Ala Gln Ala Ser Thr Asn Asn Ala Gln Val 530 535 540 Leu Ala Gln Val Arg Gln Phe Leu Asn Gln 545 550 Dr. C Sonia González Blanco Representante Legal, CIGB 1