1668:
26:
387:
of its binding site, reduced thermolysin stability by 7 °C. However, while calcium binding makes a significant contribution to stabilising thermolysin, more crucial to stability is a small cluster of N-terminal domain amino acids located at the proteins surface. In particular a
280:. However thermolysin is also widely used for peptide bond formation through the reverse reaction of hydrolysis. Thermolysin is the most stable member of a family of metalloproteinases produced by various
383:
to thermolysin stability have shown that upon thermal inactivation a single calcium ion is released from the molecule. Preventing this calcium from originally binding to the molecule by
455:
Holden HM, Matthews BW (March 1988). "The binding of L-valyl-L-tryptophan to crystalline thermolysin illustrates the mode of interaction of a product of peptide hydrolysis".
365:, thermolysin does not undergo any major conformational changes until at least 70 °C. The thermal stability of members of the TLP family is measured in terms of a
179:
992:
408:(TLP-ste), results in individual reductions in stability of 7 °C (F63→T) and 6.3 °C (P69→A) and when combined a reduction in stability of 12.3 °C.
198:
550:
Trusek-Holownia A. (2003). "Synthesis of ZAlaPheOMe, the precursor of bitter dipeptide in the two-phase ethyl acetate-water system catalysed by thermolysin".
306:
28 amino acids long, a pro-peptide 204 amino acids long and the mature enzyme itself 316 amino acids in length. The signal peptide acts as a signal for
587:"A new method for the extracellular production of recombinant thermolysin by co-expressing the mature sequence and pro-sequence in Escherichia coli"
326:
and leads to autocleavage of the peptide bond linking pro and mature sequences. The mature protein is then secreted into the extracellular medium.
1334:
643:
1021:
985:
307:
425:
1387:
846:
Yagasaki, Makoto; Hashimoto, Shin-ichi (November 2008). "Synthesis and application of dipeptides; current status and perspectives".
191:
158:
978:
134:
1543:
511:"Role of calcium ions in the thermostability of thermolysin and Bacillus subtilis var. amylosacchariticus neutral protease"
1658:
1339:
768:
Eijsink VG, Veltman OR, et al. (1995). "Structural determinants of the stability of thermolysin-like proteinases".
362:
1233:
1228:
372:
temperature. At this temperature incubation for 30 minutes reduces the enzymes activity by half. Thermolysin has a
1528:
1644:
1631:
1618:
1605:
1592:
1579:
1566:
1012:
384:
379:
value of 86.9 °C, making it the most thermo stable member of the TLP family. Studies on the contribution of
152:
1538:
1492:
1435:
1349:
1166:
1009:
964:
396:(P) at amino acid position 69 contribute significantly to thermolysin stability. Changing these amino acids to
311:
219:
45:
139:
1440:
1693:
253:
203:
334:
Thermolysin has a molecular weight of 34,600 Da. Its overall structure consists of two roughly spherical
127:
1461:
1380:
1324:
1533:
358:
in structure. These two domains are connected by a central alpha helix, spanning amino acids 137–151.
286:
species. These enzymes are also termed 'neutral' proteinases or thermolysin -like proteinases (TLPs).
1005:
902:
62:
811:
Dahlquist FW, Long JW, Bigbee WL (1976). "Role of
Calcium in the thermal stability of thermolysin".
155:
1497:
339:
323:
79:
57:
1698:
1688:
1430:
871:
793:
661:
347:
265:
970:
930:
863:
828:
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639:
608:
567:
532:
472:
445:
343:
299:
246:
146:
1476:
1471:
1445:
1373:
1273:
920:
910:
855:
820:
777:
735:
696:
631:
598:
559:
522:
464:
338:
with a deep cleft running across the middle of the molecule separating the two domains. The
319:
115:
955:
1523:
1507:
1420:
1029:
891:"Determining Biophysical Protein Stability in Lysates by a Fast Proteolysis Assay, FASTpp"
673:
421:, less bitter-tasting byproduct is produced when the reaction is catalyzed by thermolysin.
242:
91:
50:
906:
1672:
1561:
1502:
960:
925:
890:
687:
Holmes MA, Matthews BW (1982). "Structure of thermolysin refined at 1.6 A resolution".
527:
510:
335:
303:
174:
740:
723:
635:
563:
1682:
1466:
1425:
700:
389:
355:
295:
875:
797:
1415:
273:
361:
In contrast to many proteins that undergo conformational changes upon heating and
915:
1639:
1574:
1410:
1344:
1174:
628:
Engineering, expression, purification, and production of recombinant thermolysin
1667:
1319:
1191:
859:
351:
277:
269:
603:
586:
1613:
1587:
418:
397:
315:
934:
867:
653:
612:
571:
789:
749:
708:
490:
Endo, S. (1962). "Studies on protease produced by thermophilic bacteria".
476:
468:
404:(A) respectively in a less stable thermolysin-like proteinase produced by
1354:
1156:
1151:
1146:
1001:
832:
536:
282:
222:
103:
824:
781:
1141:
1136:
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1111:
401:
393:
380:
122:
449:
1626:
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1101:
1096:
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1076:
1071:
1049:
1044:
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951:
249:
186:
98:
86:
74:
889:
Minde, David P.; Maurice, Madelon M.; RĂĽdiger, Stefan G. D. (2012).
1600:
1303:
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1248:
1243:
1238:
1223:
1218:
1066:
1061:
1056:
1034:
298:
proteases thermolysin is first synthesised by the bacterium as a
268:
for structural stability. Thermolysin specifically catalyzes the
1213:
1208:
1201:
1196:
1184:
1179:
302:. Thermolysin is synthesized as a pre-proenzyme consisting of a
261:
110:
25:
1369:
974:
318:
pre-prothermolysin is then processed into prothermolysin by a
630:. Biotechnology Annual Review. Vol. 13. pp. 43–64.
1365:
424:
Determining protein stability in cell lysate using the
1656:
954:
online database for peptidases and their inhibitors:
1552:
1516:
1485:
1454:
1403:
1312:
1165:
1020:
197:
185:
173:
168:
145:
133:
121:
109:
97:
85:
73:
68:
56:
44:
39:
18:
227:Bacillus thermoproteolyticus neutral proteinase
1381:
986:
8:
1388:
1374:
1366:
993:
979:
971:
722:Matthews BW, Weaver LH, Kester WR (1974).
165:
24:
963:at the U.S. National Library of Medicine
924:
914:
763:
761:
759:
739:
602:
526:
626:Inouye K, Kusano M, et al. (2007).
1663:
585:Yasukawa K, Kusano M, Inouye K (2007).
509:Tajima M, Urabe I, et al. (1976).
437:
342:of each domain is quite different, the
310:of pre-prothermolysin to the bacterial
848:Applied Microbiology and Biotechnology
669:
659:
15:
1335:Pregnancy-associated plasma protein A
7:
392:(F) at amino acid position 63 and a
528:10.1111/j.1432-1033.1976.tb10293.x
426:fast parallel proteolysis (FASTpp)
14:
724:"The conformation of thermolysin"
322:. The prosequence then acts as a
1666:
30:Crystallographic structure of
1:
741:10.1016/S0021-9258(19)42067-X
636:10.1016/S1387-2656(07)13003-9
564:10.1016/S0168-1656(03)00024-5
264:for enzyme activity and four
1340:Bone morphogenetic protein 1
916:10.1371/journal.pone.0046147
701:10.1016/0022-2836(82)90319-9
406:Bacillus stearothermophillus
258:Bacillus thermoproteolyticus
32:Bacillus thermoproteolyticus
1715:
1544:Michaelis–Menten kinetics
1167:Matrix metalloproteinases
860:10.1007/s00253-008-1590-3
164:
23:
1436:Diffusion-limited enzyme
1350:Insulin-degrading enzyme
965:Medical Subject Headings
278:hydrophobic amino acids
604:10.1093/protein/gzm031
1529:Eadie–Hofstee diagram
1462:Allosteric regulation
1325:Procollagen peptidase
1006:metalloendopeptidases
591:Protein Eng. Des. Sel
1539:Lineweaver–Burk plot
417:In the synthesis of
312:cytoplasmic membrane
907:2012PLoSO...746147M
825:10.1021/bi00650a024
782:10.1038/nsb0595-374
492:J. Ferment. Technol
469:10.2210/pdb3tmn/pdb
346:consists of mostly
340:secondary structure
324:molecular chaperone
294:Like all bacterial
1498:Enzyme superfamily
1431:Enzyme promiscuity
348:beta pleated sheet
260:. It requires one
1654:
1653:
1363:
1362:
770:Nat. Struct. Biol
734:(24): 8030–8044.
645:978-0-444-53032-5
354:domain is mostly
344:N-terminal domain
247:metalloproteinase
213:
212:
209:
208:
128:metabolic pathway
1706:
1671:
1670:
1662:
1534:Hanes–Woolf plot
1477:Enzyme activator
1472:Enzyme inhibitor
1446:Enzyme catalysis
1390:
1383:
1376:
1367:
1030:Alpha secretases
995:
988:
981:
972:
939:
938:
928:
918:
886:
880:
879:
843:
837:
836:
819:(5): 1103–1111.
808:
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801:
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743:
719:
713:
712:
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547:
541:
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480:
452:
442:
320:signal peptidase
252:produced by the
166:
28:
16:
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1713:
1709:
1708:
1707:
1705:
1704:
1703:
1679:
1678:
1677:
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1657:
1655:
1650:
1562:Oxidoreductases
1548:
1524:Enzyme kinetics
1512:
1508:List of enzymes
1481:
1450:
1421:Catalytic triad
1399:
1394:
1364:
1359:
1308:
1161:
1016:
999:
947:
942:
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883:
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767:
766:
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721:
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646:
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620:
584:
583:
579:
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515:Eur. J. Biochem
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503:
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414:
378:
371:
332:
292:
35:
12:
11:
5:
1712:
1710:
1702:
1701:
1696:
1691:
1681:
1680:
1676:
1675:
1652:
1651:
1649:
1648:
1635:
1622:
1609:
1596:
1583:
1570:
1556:
1554:
1550:
1549:
1547:
1546:
1541:
1536:
1531:
1526:
1520:
1518:
1514:
1513:
1511:
1510:
1505:
1500:
1495:
1489:
1487:
1486:Classification
1483:
1482:
1480:
1479:
1474:
1469:
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1449:
1448:
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1428:
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1418:
1413:
1407:
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1395:
1393:
1392:
1385:
1378:
1370:
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1360:
1358:
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1352:
1347:
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1337:
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1327:
1322:
1316:
1314:
1310:
1309:
1307:
1306:
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1296:
1291:
1286:
1281:
1276:
1271:
1266:
1261:
1256:
1251:
1246:
1241:
1236:
1231:
1226:
1221:
1216:
1211:
1206:
1205:
1204:
1199:
1189:
1188:
1187:
1182:
1171:
1169:
1163:
1162:
1160:
1159:
1154:
1149:
1144:
1139:
1134:
1129:
1124:
1119:
1114:
1109:
1104:
1099:
1094:
1089:
1084:
1079:
1074:
1069:
1064:
1059:
1054:
1053:
1052:
1047:
1042:
1037:
1026:
1024:
1018:
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958:
946:
945:External links
943:
941:
940:
901:(10): e46147.
881:
838:
803:
776:(5): 374–379.
755:
714:
695:(4): 623–639.
679:
670:|journal=
644:
618:
597:(8): 375–383.
577:
558:(2): 153–163.
542:
521:(1): 243–247.
501:
482:
463:(7): 3256–60.
436:
434:
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331:
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304:signal peptide
291:
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1694:Zinc proteins
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1506:
1504:
1503:Enzyme family
1501:
1499:
1496:
1494:
1491:
1490:
1488:
1484:
1478:
1475:
1473:
1470:
1468:
1467:Cooperativity
1465:
1463:
1460:
1459:
1457:
1453:
1447:
1444:
1442:
1439:
1437:
1434:
1432:
1429:
1427:
1426:Oxyanion hole
1424:
1422:
1419:
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1133:
1130:
1128:
1125:
1123:
1120:
1118:
1115:
1113:
1110:
1108:
1105:
1103:
1100:
1098:
1095:
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1085:
1083:
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1078:
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1070:
1068:
1065:
1063:
1060:
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1055:
1051:
1048:
1046:
1043:
1041:
1038:
1036:
1033:
1032:
1031:
1028:
1027:
1025:
1023:
1022:ADAM proteins
1019:
1014:
1011:
1007:
1003:
996:
991:
989:
984:
982:
977:
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973:
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839:
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826:
822:
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804:
799:
795:
791:
787:
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779:
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751:
747:
742:
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733:
729:
728:J. Biol. Chem
725:
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629:
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619:
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553:
552:J. Biotechnol
546:
543:
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520:
516:
512:
505:
502:
497:
493:
486:
483:
478:
474:
470:
466:
462:
458:
457:J. Biol. Chem
451:
447:
441:
438:
432:
427:
423:
420:
416:
415:
411:
409:
407:
403:
399:
395:
391:
390:phenylalanine
386:
382:
375:
368:
364:
359:
357:
356:alpha helical
353:
349:
345:
341:
337:
329:
327:
325:
321:
317:
313:
309:
308:translocation
305:
301:
300:pre-proenzyme
297:
296:extracellular
289:
287:
285:
284:
279:
275:
274:peptide bonds
271:
267:
263:
259:
255:
254:Gram-positive
251:
248:
244:
240:
236:
235:thermoase Y10
232:
228:
224:
221:
217:
205:
202:
200:
196:
193:
190:
188:
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181:
178:
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160:
157:
154:
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148:
144:
141:
138:
136:
132:
129:
126:
124:
120:
117:
114:
112:
108:
105:
104:NiceZyme view
102:
100:
96:
93:
90:
88:
84:
81:
78:
76:
72:
67:
64:
61:
59:
55:
52:
49:
47:
43:
38:
33:
27:
22:
17:
1640:Translocases
1637:
1624:
1611:
1598:
1585:
1575:Transferases
1572:
1559:
1416:Binding site
1329:
1175:Collagenases
898:
894:
884:
854:(1): 13–22.
851:
847:
841:
816:
813:Biochemistry
812:
806:
773:
769:
731:
727:
717:
692:
689:J. Mol. Biol
688:
682:
627:
621:
594:
590:
580:
555:
551:
545:
518:
514:
504:
495:
491:
485:
460:
456:
440:
412:Applications
405:
373:
366:
363:denaturation
360:
350:, while the
333:
293:
281:
266:calcium ions
257:
243:thermostable
238:
234:
230:
226:
215:
214:
92:BRENDA entry
34:thermolysin.
31:
1411:Active site
1345:Lysostaphin
1330:Thermolysin
1192:Gelatinases
961:Thermolysin
276:containing
216:Thermolysin
80:IntEnz view
40:Identifiers
19:Thermolysin
1683:Categories
1614:Isomerases
1588:Hydrolases
1455:Regulation
1320:Neprilysin
498:: 346–353.
433:References
352:C-terminal
270:hydrolysis
149:structures
116:KEGG entry
63:9073-78-3
1699:Proteases
1689:EC 3.4.24
1493:EC number
1002:Proteases
672:ignored (
662:cite book
453:;
419:aspartame
398:threonine
330:Structure
316:periplasm
314:. In the
290:Synthesis
256:bacteria
231:thermoase
223:3.4.24.27
69:Databases
51:3.4.24.27
1517:Kinetics
1441:Cofactor
1404:Activity
1355:ZMPSTE24
1157:ADAMTS13
1152:ADAMTS12
1147:ADAMTS10
935:23056252
895:PLOS ONE
876:10200090
868:18795289
798:37785818
654:17875473
613:17616558
572:12697393
400:(T) and
385:mutation
283:Bacillus
262:zinc ion
245:neutral
204:proteins
192:articles
180:articles
153:RCSB PDB
1673:Biology
1627:Ligases
1397:Enzymes
1142:ADAMTS9
1137:ADAMTS8
1132:ADAMTS5
1127:ADAMTS4
1122:ADAMTS3
1117:ADAMTS2
1112:ADAMTS1
956:M04.001
926:3463568
903:Bibcode
790:7664094
750:4214815
709:7175940
477:3343246
402:alanine
394:proline
381:calcium
336:domains
241:) is a
140:profile
123:MetaCyc
58:CAS no.
1659:Portal
1601:Lyases
1279:MMP23B
1274:MMP23A
1107:ADAM33
1102:ADAM28
1097:ADAM23
1092:ADAM22
1087:ADAM18
1082:ADAM15
1077:ADAM12
1072:ADAM11
1050:ADAM19
1045:ADAM17
1040:ADAM10
1013:3.4.24
967:(MeSH)
952:MEROPS
933:
923:
874:
866:
833:814920
831:
796:
788:
748:
707:
652:
642:
611:
570:
537:819262
535:
475:
428:assay.
250:enzyme
187:PubMed
169:Search
159:PDBsum
99:ExPASy
87:BRENDA
75:IntEnz
46:EC no.
1553:Types
1313:Other
1304:MMP28
1299:MMP27
1294:MMP26
1289:MMP25
1284:MMP24
1269:MMP21
1264:MMP20
1259:MMP19
1254:MMP17
1249:MMP16
1244:MMP15
1239:MMP14
1234:MMP13
1229:MMP12
1224:MMP11
1219:MMP10
1067:ADAM8
1062:ADAM7
1057:ADAM2
1035:ADAM9
872:S2CID
794:S2CID
135:PRIAM
1645:list
1638:EC7
1632:list
1625:EC6
1619:list
1612:EC5
1606:list
1599:EC4
1593:list
1586:EC3
1580:list
1573:EC2
1567:list
1560:EC1
1214:MMP7
1209:MMP3
1202:MMP9
1197:MMP2
1185:MMP8
1180:MMP1
950:The
931:PMID
864:PMID
829:PMID
786:PMID
746:PMID
705:PMID
674:help
650:PMID
640:ISBN
609:PMID
568:PMID
533:PMID
473:PMID
450:3TMN
199:NCBI
156:PDBe
111:KEGG
921:PMC
911:doi
856:doi
821:doi
778:doi
736:doi
732:249
697:doi
693:160
632:doi
599:doi
560:doi
556:102
523:doi
465:doi
461:263
446:PDB
272:of
239:TLN
175:PMC
147:PDB
1685::
1010:EC
1004::
929:.
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909:.
897:.
893:.
870:.
862:.
852:81
850:.
827:.
817:15
815:.
792:.
784:.
772:.
758:^
744:.
730:.
726:.
703:.
691:.
666::
664:}}
660:{{
648:.
638:.
607:.
595:20
593:.
589:.
566:.
554:.
531:.
519:64
517:.
513:.
496:40
494:.
471:.
459:.
448::
377:50
370:50
237:,
233:,
229:,
225:,
220:EC
1661::
1647:)
1643:(
1634:)
1630:(
1621:)
1617:(
1608:)
1604:(
1595:)
1591:(
1582:)
1578:(
1569:)
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994:e
987:t
980:v
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479:.
467::
374:T
367:T
218:(
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