WT Val QGQYKNQDPI ILVHGFNGFT DDINPAVLAH YWGGDKLNIR QDLESNGYET
....|....| ....|....| ....|....| ....|....| ....|....|
10 20 30 40 50
WT_Val QGQYKNQDPI ILVHGFNGFT DDINPAVLAH YWGGDKLNIR QDLESNGYET
Mut_Lys QGQYKNQDPI ILVHGFNGFT DDINPAVLAH YWGGDKLNIR QDLESNGYET
Mut_Asp QGQYKNQDPI ILVHGFNGFT DDINPAVLAH YWGGDKLNIR QDLESNGYET
Clustal Co ********** ********** ********** ********** **********
....|....| ....|....| ....|....| ....|....| ....|....|
60 70 80 90 100
WT_Val YEASVGALSS NYDRAVELYY YIKGGTVDYG AAHAEKYGHE RYGKTYEGVY
Mut_Lys YEASVGALSS NYDRAVELYY YIKGGTVDYG AAHAEKYGHE RYGKTYEGVY
Mut_Asp YEASVGALSS NYDRAVELYY YIKGGTVDYG AAHAEKYGHE RYGKTYEGVY
Clustal Co ********** ********** ********** ********** **********
....|....| ....|....| ....|....| ....|....| ....|....|
110 120 130 140 150
WT_Val KDWQPGKKVH LVAHSMGGQT VRQLEELLRN GNQEEIEYQK EHGGEISPLF
Mut_Lys KDWQPGKKVH LVAHSMGGQT VRQLEELLRN GNQEEIEYQK EHGGEISPLF
Mut_Asp KDWQPGKKVH LVAHSMGGQT VRQLEELLRN GNQEEIEYQK EHGGEISPLF
Clustal Co ********** ********** ********** ********** **********
....|....| ....|....| ....|....| ....|....| ....|....|
160 170 180 190 200
WT_Val QGNNDNMVNS ITTIGTPHNG THAADALGNE AIVRQLAFDY AKFKGNKNSK
Mut_Lys QGNNDNMVNS ITTIGTPHNG THAADALGNE AIVRQLAFDY AKFKGNKNSK
Mut_Asp QGNNDNMVNS ITTIGTPHNG THAADALGNE AIVRQLAFDY AKFKGNKNSK
Clustal Co ********** ********** ********** ********** **********
....|....| ....|....| ....|....| ....|....| ....|....|
210 220 230 240 250
WT_Val VDFGFGQWGL KQREGETYAQ YVQRVQNSGL WKTEDNGFYD LTREGAAKLN
Mut_Lys VDFGFGQWGL KQREGETYAQ YVQRVQNSGL WKTEDNGFYD LTREGAAKLN
Mut_Asp VDFGFGQWGL KQREGETYAQ YVQRVQNSGL WKTEDNGFYD LTREGAAKLN
Clustal Co ********** ********** ********** ********** **********
....|....| ....|....| ....|....| ....|....| ....|....|
260 270 280 290 300
WT_Val KNTSLNPNIV YKTYTGESTR PTLFGNQKSD VNLFLPFTVT GNVIGKAAEK
Mut_Lys KNTSLNPNIV YKTYTGESTR PTLFGNQKSD VNLFLPFTVT GNVIGKAAEK
Mut_Asp KNTSLNPNIV YKTYTGESTR PTLFGNQKSD VNLFLPFTVT GNVIGKAAEK
Clustal Co ********** ********** ********** ********** **********
....|....| ....|....| ....|....| ....|....| ....|....|
310 320 330 340 350
WT_Val EWRENDGLVS TISSQHPFNQ AFIEATDEVK KGVWQVTPIK HGWDHVDFVG
Mut_Lys EWRENDGLKS TISSQHPFNQ AFIEATDEVK KGVWQVTPIK HGWDHVDFVG
Mut_Asp EWRENDGLDS TISSQHPFNQ AFIEATDEVK KGVWQVTPIK HGWDHVDFVG
Clustal Co ******** * ********** ********** ********** **********
....|....| .
360
WT_Val QDSTDSNHPT E
Mut_Lys QDSTDSNHPT E
Mut_Asp QDSTDSNHPT E
Clustal Co ********** *
Figure 1. Aminoacid alignement of S. xylosus recombinant lipase (WT-Val) and mutants (Mut-Asp and Mut-Lys). Catalytic triade was shown is red, mutated aminoacids were underlined.
Figure 2. SDS-PAGE of recombinant S. xylosus lipases purification by nickel resin. Lane 1 Molecular weight markers, lane 2, BSA;lanes 3 and 4,WT-Lip;lanes 5 and 6, Mut-Asp;lanes 7 and 8 Mut-Lys.