{"refrec":{"BRefID":223230,"RR":"<b>Toogood, H.S.; Hollingsworth, E.J.; Brown, R.C.; Taylor, I.N.; Taylor, S.J.C.; McCague, R.; Littlechild, J.A.</b> (2002). A thermostable L-aminoacylase from <i>Thermococcus litoralis</i>: cloning, overexpression, characterization, and applications in biotransformations. <i>Extremophiles 6(2)</i>: 111-122. <a href=\"http://dx.doi.org/10.1007/s007920100230\" target=\"_blank\">http://dx.doi.org/10.1007/s007920100230</a>","BEntID":214954,"PublicFlag":1,"CheckedFlag":0,"wosflag":1,"vabbflag":null,"RefStringPartII":". <i>Extremophiles 6(2)</i>: 111-122. <a href=\"http://dx.doi.org/10.1007/s007920100230\" target=\"_blank\">http://dx.doi.org/10.1007/s007920100230</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Toogood, H.S.; Hollingsworth, E.J.; Brown, R.C.; Taylor, I.N.; Taylor, S.J.C.; McCague, R.; Littlechild, J.A.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Toogood, H.S. <i>et al.</i>","Englishabstract":"A thermostable L-aminoacylase from Thermococcus litoralis was cloned, sequenced, and overexpressed in Escherichia coli. The enzyme is a homotetramer of 43 kDa monomers and has an 82% sequence identity to an aminoacylase from Pyrococcus horikoshii and 45% sequence identity to a carboxypeptidase from Sulfolobus solfataricus. It contains one cysteine residue that is highly conserved among aminoacylases. Cell-free extracts of the recombinant enzyme were characterized and were found to have optimal activity at 85°C in Tris-HCl at pH 8.0. The recombinant enzyme is thermostable, with a half-life of 25 h at 70°C. Aminoacylase inhibitors, such as mono-tert-butyl malonate, had only a slight effect on activity. The enzyme was partially inhibited by EDTA and p- hydroxymercuribenzoate, suggesting that the cysteine residue and a metal ion are important, but not essential, for activity. Addition of Zn2+ and Co2+ to the apoenzyme increased the enzyme activity, whereas Sn4+ and Cu2+ almost completely abolished enzyme activity. The enzyme was most specific for substrates containing N-benzoyl- or N-chloroacetyl-amino acids, preferring substrates containing hydrophobic, uncharged, or weakly charged amino acids such as phenylalanine, methionine, and cysteine.","AbstractOtherLang":null,"BibLvlCode":"AS","StandardTitle":"A thermostable L-aminoacylase from <i>Thermococcus litoralis</i>: cloning, overexpression, characterization, and applications in biotransformations","OrigTitleLangCode":"en","OrigTitleLangCodeExtended":"eng","OrigTitleLangID":15,"DateLastModified":{"date":"2024-12-10 01:33:01.897972","timezone_type":1,"timezone":"+01:00"},"UserAccessRight":null,"UserAccID":null,"AuthorKeywords":null,"OtherDescriptors":null,"Notes":null,"AnaPub":2002,"MonPub":null,"DateUpdate":"2013-02-27","DateCreate":"2013-02-27","SecASFANote":null,"ConfID":null,"PeerRev":1,"VlizCoreFlag":1,"WoScode":"WOS:000175220800004","VABBcode":null,"OpenAcc":0,"DOI":"10.1007/s007920100230"},"refs":null,"anarec":{"AnaID":223230,"PubliDate":2002,"Pagination":"111-122","XtraPublOfAnaID":null,"ISBN":null,"Volume":"6","Issue":"2","BRefMon":null,"BRefMonRR":null,"BRefXtra":null,"BRefXtraRR":null,"SerBRefID":218588,"SerRR":"Extremophiles. 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