Asp371 / Ala371 / Asp371 / Ala371 / Asp371 / Asp371 / Ala371
Arg34 (interact with Leu33 which involved in channel leading towards the active site) / - / - / - / + / + / +
Pro368 / +
Ser369 / +
Phe370 / +
Ile372 / +
Arg373 / +
Ala374 / +
Phe375 / + / Ala375 / + / Ala375 / + / Ala375
Phe375 / Phe375 / Ala375 / Phe375 / Ala375 / Phe375 / Ala375
Ile161 / + / - / + / - / + / -
Phe267 / + / - / + / - / + / -
Tyr354 (make conection with Thr269, which is close to Asp317 and with other amino acids involved in the stabilization of the serine loop and contributing to the lipase thermostability) / +
Ile361 (interacts with His112 from AHSQG and Ile362 from the active site cleft) / +
Phe370 / +
Asp371 / Ala371 / + / Ala371 / + / + / +
Ile372 / +
Arg373 / +
Ala374 / +
Tyr376 / Ala376 / Ala376 / + / + / + / +
Leu377 / +
Arg378 / +
Tyr376 / Ala376 / Ala376 / Tyr376 / Tyr376 / Ala376 / Tyr376
Leu12 (involved in oxyanion hole) / - / - / + / + / - / +
Trp30 (conservative amino acid) / - / + / + / + / - / +
Ile37 / +
Trp40 / +
Leu41 (conservative amino acid) / +
Ile110 (interacts with Ala111 from AHSQG) / - / - / + / + / + / +
- / - / - / His112 (involved in AHSQG pentapeptide; thermostability) / - / - / -
Ile161 / - / - / + / + / - / +
Ile361 (interacts with His112 from AHSQG and Ile362 from the active site cleft) / - / - / + / + / - / +
Ile362 (active site cleft; acyl chain binding pocket) / - / - / + / + / - / +
Phe370 / - / - / + / + / - / +
Ile372 / +
Arg373 / +
Ala374 / +
Phe375 / + / Ala375 / + / Ala375 / + / +
Leu377 / +
Arg378 / +
Interaction of other important amino acids in GD-95-10, GD-95-20 lipases and R1-R6 mutants
Phe16 (stabilization of the serine loop; involved in oxyanion hole; active site cleft) / Phe16 / Phe16 / Phe16 / Phe16 / Phe16 / Phe16 / Phe16
Gly15 (involved in oxyanion hole) / +
Thr17 (involved in oxyanion hole) / +
Tyr29 (active site cleft) / - / - / + / + / - / - / +
Pro55 (binding of substrate) / +
Leu56 (active site cleft) / +
Arg62 (stabilizing role during the opening of the lid) / +
Ser113 (active site) / +
Gln114 (stabilization of the serine loop) / +
Pro164(The active site cleft) / +
Leu170 (acyl chain binding pocket) / +
Phe180 (active site cleft) / +
Leu183 (acyl chain binding pocket) / +
Gln184 (involved in lid domain) / +
Val187 (active site cleft) / + / - / + / - / + / + / +
Tyr204 (active site cleft) / +
Ala240 (active site cleft) / +
Leu244 (active site cleft) / +
Ile319 (involved in packing of catalytic Ser113) / +
Val320 (acyl chain binding pocket) / +
His358 (active site) / +
Ser113 (active site) / Ser113 / Ser113 / Ser113 / Ser113 / Ser113 / Ser113 / Ser113
Gly15 (involved in oxyanion hole) / +
Phe16 (involved in oxyanion hole; active site cleft) / +
- / Thr17 (involved in oxyanion hole) / - / - / - / + / - / -
Tyr29 (active site cleft) / +
His112 (involved in AHSQG pentapeptide; thermostability) / +
Gln114 (involved in AHSQG pentapeptide) / +
Gly115 (involved in AHSQG pentapeptide) / +
Gly116 / +
Ile161 / +
Ala162 / +
Thr163 / +
Pro164 (active site cleft) / +
- / Val320 (acyl chain binding pocket) / - / - / + / + / - / +
His358 (active site) / +
Gln114(stabilization of the serine loop) / Gln114 / Gln114 / Gln114 / Gln114 / Gln114 / Gln114 / Gln114
Gly15 (involved in oxyanion hole) / +
Phe16 (involved in oxyanion hole; active site cleft) / +
Pro55 (binding of substrate) / +
Ser57 (stabilizing role during the opening of the lid) / - / + / + / + / + / + / +
Asn59 / +
Arg62 (stabilizing role during the opening of the lid) / +
His112 (involved in AHSQG pentapeptide; thermostability) / +
Ser113 (active site) / +
Gly115 (involved in AHSQG pentapeptide) / +
Gly116 / +
Gln117 / +
Thr118 / +
Thr163 / +
Pro164(The active site cleft) / +
Ala240 (active site cleft) / +
Asp243 / +
Leu244 (active site cleft) / +
Leu170 (acyl chain binding pocket) / Leu170 / Leu170 / Leu170 / Leu170 / Leu170 / Leu170 / Leu170
Phe16 (involved in oxyanion hole; active site cleft) / + / + / + / + / - / + / +
Thr168 / +
Thr169 / +
Val171 (active site cleft) / +
Asn172 / +
Met173 (acyl chain binding pocket) / +
Phe176 (active site cleft) / +
Arg179 (involved in lid domain) / + / + / + / - / + / + / +
Phe180 (active site cleft) / +
Leu183 (acyl chain binding pocket) / +
Leu244 (active site cleft) / + / - / + / + / + / + / +
Cys295 / +
Phe298 (active site cleft) / +
Leu299 / +
Ile319 (involved in packing of catalytic Ser113) / +
Ala240 (active site cleft) / Ala240 / Ala240 / Ala240 / Ala240 / Ala240 / Ala240 / Ala240
Phe16 (involved in oxyanion hole; active site cleft) / +
Pro55 (interaction with substrate) / +
Leu56 (active site cleft) / +
Ser57 (stabilizing role during the opening of the lid) / +
Ser58 / +
- / Asn59 / - / + / + / - / - / +
Arg62 (stabilizing role during the opening of the lid) / + / - / + / + / + / + / +
Gln114 (involved in AHSQG pentapeptide) / +
Phe180 (active site cleft) / +
Trp234 / + / - / - / + / - / - / -
Thr239 / +
Arg241 / +
Tyr242 / +
Asp243 / +
Leu244 (active site cleft) / +
Asp317 (active site) / Asp317 / Asp317 / Asp317 / Asp317 / Asp317 / Asp317 / Asp317
Ala162 / +
Thr269 (involved in packing of catalytic Ser113) / +
Glu270 / +
Arg271 / +
Met287 / +
Asn288 / - / + / + / + / - / - / +
Ser291 / - / - / - / + / + / + / +
- / Asn316 / + / + / + / + / + / +
- / Gly318 / + / + / + / + / + / +
Ile319 (involved in packing of catalytic Ser113) / +
Val320 (acyl chain binding pocket) / +
Thr322 / +
Met325 (involved in packing of catalytic Ser113) / +
Val356 / +
Asp357 / +
His358 (active site) / +
- / - / - / - / - / Leu359 (acyl chain-binding pocket) / - / -
Ile319 (involved in packing catalytic Ser113) / Ile319 / Ile319 / Ile319 / Ile319 / Ile319 / Ile319 / Ile319
Phe16 (involved in oxyanion hole; active site cleft) / +
Thr168 / +
Leu170 (acyl chain binding pocket) / +
Leu183(acyl chain binding pocket) / +
Leu244 (active site cleft) / +
- / - / Asn288 / + / - / - / - / +
Phe290(acyl chain binding pocket) / - / + / - / - / + / - / +
Ser291 / +
Cys295 / +
Leu299 / +
Asp317 (active site) / +
Gly318 / +
Val320(acyl chain binding pocket) / +
- / Asp357 / - / - / - / - / + / -
His358 (active site) / +
Leu359 (acyl chain-binding pocket) / +
Val320 (active site cleft; acyl chain binding pocket) / Val320 / Val320 / Val320 / Val320 / Val320 / Val320 / Val320
Phe16 (involved in oxyanion hole; active site cleft) / +
- / Ser113 (active site) / - / + / + / + / - / +
Ala162 / +
Thr163 / +
Pro164 (active site cleft) / +
Gly167 / +
Thr168 / +
Leu244 (active site cleft) / +
Leu299 / +
Asn316 / +
Asp317 (active site) / +
Gly318 / +
Ile319 (involved in packing catalytic Ser113) / +
Asn321 / +
Thr322 / +
Ser324 / +
- / Met325 (involved in packing of catalytic Ser113) / + / + / + / + / + / +
His358 (active site) / +
Tyr354(make conection with Thr269, which is close to Asp317 and with other amino acids involved in the stabilization of the serine loop and contributing to the lipase thermostability) / Tyr354 / Tyr354 / Tyr354 / Tyr354 / Tyr354 / Tyr354 / Tyr354
Phe267 / +
Ser268 / +
Thr269 (involved in packing of catalytic Ser113) / +
Glu270 / +
Met351 / +
Gly352 / +
Thr353 / +
Asn355 / +
Val356 / +
Ile361 (interacts with His112 from AHSQG and Ile362 from the active site cleft) / +
Phe370 / + / + / + / + / + / + / -
Phe375 / + / + / + / + / + / + / -
Arg378 / + / + / + / + / + / + / -
His358 (active site) / His358 / His358 / His358 / His358 / His358 / His358 / His358
Phe16 (involved in oxyanion hole; active site cleft) / +
Tyr29 (active site cleft) / + / + / + / + / + / - / +
His112 (involved in AHSQG pentapeptide; thermostability) / +
Ser113 (active site) / +
Ala162 / +
Pro164 (active site cleft) / +
Thr269 (involved in packing of catalytic Ser113) / +
Asn288 / - / + / + / + / - / - / +
Ser291 / - / - / - / + / + / - / -
Asp317 (active site) / + / + / + / + / + / + / +
Gly318 / - / + / - / + / + / - / +
Ile319 (involved in packing catalytic Ser113) / +
Val320 (acyl chain binding pocket) / +
Asp357 / +
Leu359 (acyl chain-binding pocket) / +
Glu360 (binding of Ca2+) / +
Ile361 (interacts with His112 from AHSQG and Ile362 from the active site cleft) / +
Ile362 (active site cleft. acyl chain binding pocket) / +
Leu359 (acyl chain-binding pocket) / Leu359 / Leu359 / Leu359 / Leu359 / Leu359 / Leu359 / Leu359
- / - / Thr17 (involved in oxyanion hole) / - / - / - / - / +
- / - / Tyr29 (active site cleft) / - / - / - / - / +
- / - / His112 (involved in AHSQG pentapeptide; thermostability) / - / - / - / - / +
Leu183 (acyl chain binding pocket) / +
Ala186 (involved in lid domain) / +
Val187 (active site cleft) / +
Ala190 (substrate binding cleft) / + / + / + / + / + / + / -
Asn288 / +
Phe290 (acyl chain binding pocket) / - / + / + / + / + / - / +
Ser291 / + / - / + / + / + / + / -
- / - / - / - / - / Asp317 (active site) / - / -
Ile319 (involved in packing catalytic Ser113) / +
Asp357 / +
His358 (active site) / +
Glu360 (interactions with Ca2+) / +
Ile361 (interacts with His112 from AHSQG and Ile362 from the active site cleft) / +
Ile362 (active site cleft. acyl chain binding pocket) / +
Gly363 / +
Val364 (active site cleft) / +
Asp365 / +
Ile361 (interacts with His112 from AHSQG and Ile362 from the active site cleft) / Ile361 / Ile361 / Ile361 / Ile361 / Ile361 / Ile361 / Ile361
- / Trp30 (conservative amino acid) / + / - / - / + / - / +
- / - / - / Arg34 (interact with Leu33 which involved in channel leading towards the active site) / - / - / - / -
- / - / - / Ile37 / + / + / + / +
His112 (involved in AHSQG pentapeptide; thermostability) / +
Ile161 / +
Ala162 / +
Phe267 / +
Ser268 / - / - / - / - / + / - / -
Thr269 (involved in packing of catalytic Ser113) / +
Tyr354 (make conection with Thr269, which is close to Asp317 and with other amino acids involved in the stabilization of the serine loop and contributing to the lipase thermostability) / +
Val356 / +
Asp357 / +
His358 (active site) / +
Leu359(acyl chain-binding pocket) / +
Glu360 (binding of Ca2+) / +
Ile362 (active site cleft; acyl chain binding pocket) / +
Gly363 / +
Phe370 / + / + / + / + / + / + / -
Ile372 / + / + / + / + / + / + / -
Phe375 / + / + / + / + / + / + / -
Tyr376 / - / - / + / + / - / + / -
Ile362 (active site cleft. acyl chain binding pocket) / Ile362 / Ile362 / Ile362 / Ile362 / Ile362 / Ile362 / Ile362
Thr17 (involved in oxyanion hole) / - / + / + / + / + / + / +
Tyr29 (active site cleft) / +
Trp30 (conservative amino acid) / +
- / Gly31 / + / + / + / + / + / +
Gly32 / +
- / Arg34 (interact with Leu33 which involved in channel leading towards the active site) / - / + / - / - / - / +
Ile37 / +
His112 (involved in AHSQG pentapeptide; thermostability) / +
Ile161 / - / - / - / - / - / - / +
His358 (active site) / +
Leu359 (acyl chain-binding pocket) / +
Glu360 (interactions with Ca2+) / +
Ile361 (interacts with His112 from AHSQG and Ile362 from the active site cleft) / +
Gly363 / +
Val364 (active site cleft) / +
Ile372 / + / + / + / + / + / + / -
Tyr376 / - / - / + / + / - / + / -
Val364(active site cleft) / Val364 / Val364 / Val364 / Val364 / Val364 / Val364 / Val364
- / - / - / - / Thr17 (involved in oxyanion hole) / - / + / -
Trp19 (acyl chain-binding pocket) / +
Phe27 (active site cleft) / +
Lys28 / +
Tyr29 (active site cleft) / +
Gly32 / +
Val33 / +
Val187 (active site cleft) / +
Ala190 (substrate binding cleft) / +
- / Asn288 / - / - / - / + / + / -
Leu359 (acyl chain-binding pocket) / +
- / - / Glu360 (binding of Ca2+) / - / + / - / + / -
Ile362 (active site cleft. acyl chain binding pocket) / +
Gly363 / +
Asp365 (interactions with Ca2+) / +
Pro366 (interactions with Ca2+) / +
Table S2 Interactions changes of important amino acids in mutated lipase structures. + - contact detectable; - relationships not detectable.The differences are greyed. The importance of amino acids, with wich the interactions are formed, predicted according [22-24].