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.
B7T0E8: Gp58
P10547: Lysostaphin
UniProt

IPR007921:15-149
IPR013338:483-629
IPR003646:278-367
IPR003646:411-481
InterPro

GM0010:1-634
GM0010:635-884
GMEnzy

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493

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

Domains and repeats

15-149:  IPR007921, the CHAP domain
483-629:  IPR013338, the Lysozyme domain, subfamily 2

GO term prediction

Biological Process:  0009253, the peptidoglycan catabolic process
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
GM0010[635-884]: derive from protein P10547 sequence 1-246

P10547, 493 AA., the Lysostaphin from Staphylococcus simulans

Source: Staphylococcus simulans

Domains and repeats

239-382:  IPR011055, the Duplicated hybrid motif?
411-481:  IPR003646, the SH3-like domain, bacterial-type
278-367:  IPR003646, the Peptidase M23?

GO term prediction

    No GO info found on GM0010

Linking to UniprotKB
Linging to InterPro

Annotation

1-634
635-884
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1127

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
  BlastP to GMEnzy

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 assays
  showed 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]

Comments

    No comments found on GM0010


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