Detail of GM0010
Summary
The genetically modified enzybiotic, named HydH5Lyso , constructed by Domains Assembly strategy, improve the lytic activity.With HydH5Lyso
Sequence Length: 879 AA.
Mass: 99390.8 Da.
Isoelectric Point: 9.96
Function: HydH5Lyso and HydH5SH3b, showed a 2.5-fold higher specific activity than the parental protein HydH5
Construction
GM0010, constructed by Domains Assembly strategy. No schema construted on GM0010
Details:
GM0010[1-634]: derive from protein B7T0E8 sequence 1-634
B7T0E8, 634 AA., the Gp58 from Staphylococcus phage phiSauS-IPLA88
Source: Staphylococcus phage phiSauS-IPLA88
483-629: IPR013338, the Lysozyme domain, subfamily 2
Biological Process: 0044036, the cell wall macromolecule metabolic process
Molecular Function: 0004040, the amidase activity?
Molecular Function: 0016787, the hydrolase activity?
Linking to UniprotKB
Linging to InterPro
Domains and repeats
15-149: IPR007921, the CHAP domain483-629: IPR013338, the Lysozyme domain, subfamily 2
GO term prediction
Biological Process: 0009253, the peptidoglycan catabolic processBiological Process: 0044036, the cell wall macromolecule metabolic process
Molecular Function: 0004040, the amidase activity?
Molecular Function: 0016787, the hydrolase activity?
Linking to UniprotKB
Linging to InterPro
GM0010[635-884]: derive from protein P10547 sequence 1-246
P10547, 493 AA., the Lysostaphin from Staphylococcus simulans
Annotation
1-634
635-884
15-149
: EAD, CHAP domain, derived from phage vB_SauS-phiIPLA88 (phiIPLA88)PG hydrolase(HydH5);483-629
: EAD, LYZ2 domain, derived from phage vB_SauS-phiIPLA88 (phiIPLA88)PG hydrolase(HydH5);667-767
: EAD, Endopeptidase domain, derived from lysostaphin;802-884
: CBD, SH3b domain, derived from lysostaphin;Domain and repeats:
15-149
: IPR007921,CHAP domain; 483-629
: IPR013338,Lysozyme domain, subfamily 2; GO term prediction:
0016787, the hydrolase activity
MGLPNPKDRKPTASEVVEWALYMAKNRRVIDVDRSYGGQCWDVPNYILERYWGFRTWGNANAMAQKSNYRGRDFKIYRNTASFTPKPGDWAVWANRNPGHVAIVVGPADKNAFVSVDQNWYTANWSGSPPYKIKHTYHDGPGGVTHFVRPPYHPDKTTPAPQPVPKPKDDSDDKEKNNKKVPIWKDVKTIKYTISSQVVNYPEYIYHFIVEGNRRLEKPKGIMIRNAQTMSSVENLYNSRKKYKQDVEYPHFYVDRHNIWAPRRAVFEVPNEPDYIVIDVCEDYSASKNEFIFNEIYAMGVAVDMMVEYEIPLSIENLKVDDSIWRSMLEHVNWNMIDNGVPPKDKYEALEKALLNIFKNREKLLNSITKPTVTKSRIKVMVDNKNADIANVRDSSPTANNGSASKQPQIITETSPYTFKQALDRQMSRGNPKKSHTWGWANATRAQTSSSMNVKRIWESNTQCYQMLNLGKYQGVSVSSLNKILKGKGTLNNQGKAFAEACKKHNINEIYLIAHAFLESGYGTSNFASGKDGVYNYFGIGAYDNNPNYAMTFARNKGWTSPAKAIMGGASFVRKDYINKGQNTLYRIRWNPKNPATHQYATAIEWCQHQASTIAKLYKQIGLKGVYFTRDKYKLE-ALRAATHEHSAQWLNNYKKGYGYGPYPLGINGGMHYGVDFFMNIGTPVKAISSGKIVEAGWSNYGGGNQIGLIENDGVHRQWYMHLSKYNVKVGDYVKAGQIIGWSGSTGYSTAPHLHFQRMVNSFSNSTAQDPMPFLKSAGYGKAGGTVTPTPNTGWKTNKYGTLYKSESASFTPNTDIITRTTGPFRSMPQSGVLKAGQTIHYDEVMKQDGHVWVGYTGNSGQRIYLPVRTWNKSTNTLGV
Production
1. Expressed by pET25a in E. coli BL21(DE3), Purified by NiNTA nickel column chromatography (Qiagen, Valencia, CA).Activity
1. lytic activity test on Staphylococcus aureus (live S. aureus Sa9 cells ) by the turbidity-reduction assaysshowed specific activity of 0.083 ∆OD600nm min-1µM-1
Reference
1. Lorena Rodriguez-Rubio, Beatriz Martinez, Ana Rodriguez1, David M. Donovan, and Pilar Garc¨ªa. (2012) Enhanced staphylolytic activity of the Staphylococcus aureus bacteriophage vB_SauS-phiIPLA88 HydH5 virion associated peptidoglycan hydrolase: fusions, deletions and synergy with LysH5. Appl. Environ. Microbiol.. 4431-2:32-41. [doi:10.1128/AEM.07621-11] [PMID:22267667] [FULL TEXT]2. Rodriguez-Rubio, L.Martinez, B.Donovan, D. M.Garcia, P.Rodriguez, A.. (2013) Potential of the virion-associated peptidoglycan hydrolase HydH5 and its derivative fusion proteins in milk biopreservation. PLoS One. 01:e54828. [doi:10.1371/journal.pone.0054828] [PMID:23359813] [FULL TEXT]
3. Rodriguez-Rubio L, Martinez B, Rodriguez A, Donovan DM, Gotz F,Pilar Garcia. (2013) The phage lytic proteins from the Staphylococcus aureus bacteriophage vB_SauS-phiIPLA88 display multiple active catalytic domains and do not trigger staphylococcal resistance. PLoS One . REF00455:e64671. [doi:10.1371/journal.pone.0064671.] [PMID:23724076] [FULL TEXT]