Detail of GM0008
Summary
The genetically modified enzybiotic, named HydH5SH3b , constructed by Domains Assembly strategy, improve the lytic activity.With HydH5SH3b
Sequence Length: 739 AA.
Mass: 84157.8 Da.
Isoelectric Point: 10
Function: HydH5SH3b, showed a 1.7-fold higher specific activity than the parental protein HydH5
Construction
GM0008, constructed by Domains Assembly strategy. No schema construted on GM0008
Details:
GM0008[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
GM0008[635-739]: derive from protein P10547 sequence 411-485
P10547, 493 AA., the Lysostaphin from Staphylococcus simulans
Annotation
1-634
635-739
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);637-739
: CBD, SH3b domain, derived from lysostaphin;Domain and repeats:
15-149
: IPR007921,CHAP domain; 483-629
: IPR013338,Lysozyme domain, subfamily 2; 635-735
: IPR003646,SH3-like domain, bacterial-type; GO term prediction:
0016787, the hydrolase activity
MGLPNPKDRKPTASEVVEWALYMAKNRRVIDVDRSYGGQCWDVPNYILERYWGFRTWGNANAMAQKSNYRGRDFKIYRNTASFTPKPGDWAVWANRNPGHVAIVVGPADKNAFVSVDQNWYTANWSGSPPYKIKHTYHDGPGGVTHFVRPPYHPDKTTPAPQPVPKPKDDSDDKEKNNKKVPIWKDVKTIKYTISSQVVNYPEYIYHFIVEGNRRLEKPKGIMIRNAQTMSSVENLYNSRKKYKQDVEYPHFYVDRHNIWAPRRAVFEVPNEPDYIVIDVCEDYSASKNEFIFNEIYAMGVAVDMMVEYEIPLSIENLKVDDSIWRSMLEHVNWNMIDNGVPPKDKYEALEKALLNIFKNREKLLNSITKPTVTKSRIKVMVDNKNADIANVRDSSPTANNGSASKQPQIITETSPYTFKQALDRQMSRGNPKKSHTWGWANATRAQTSSSMNVKRIWESNTQCYQMLNLGKYQGVSVSSLNKILKGKGTLNNQGKAFAEACKKHNINEIYLIAHAFLESGYGTSNFASGKDGVYNYFGIGAYDNNPNYAMTFARNKGWTSPAKAIMGGASFVRKDYINKGQNTLYRIRWNPKNPATHQYATAIEWCQHQASTIAKLYKQIGLKGVYFTRDKYKLE-GGTVTPTPNTGWKTNKYGTLYKSESASFTPNTDIITRTTGPFRSMPQSGVLKAGQTIHYDEVMKQDGHVWVGYTGNSGQRIYLPVRTWNKSTNTLGVLWGTIK
Production
1. Expressed by pET23a 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 a) by the turbidity-reduction assaysshowed specific activity of 0.056 ∆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]