Detail of GM0012


Summary

The genetically modified enzybiotic, named CHAP166 , constructed by Truncation strategy, decrease the lytic activity.
With  CHAP166
  Sequence Length:  166 AA.
  Mass:  18903.2 Da.
  Isoelectric Point:  10.12
  Function:  CHAP domain with ~ 1.4-fold reduction in specific activity (0.023 ± 0.001 ΔOD600nm min-1µM-1) observed when compared to full-length HydH5 activity.

Construction

GM0012, constructed by Truncation strategy.
B7T0E8: Gp58
UniProt

IPR007921:15-149
IPR013338:483-629
InterPro

GM0012:1-166
GMEnzy

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Details:
GM0012[1-166]: derive from protein B7T0E8 sequence 1-166

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

Annotation

1-166
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1-166
: EAD, CHAP domain, derived from Staphylococcus aureus bacteriophage vB_SauS-phi-IPLA88 endolysin HydH5;
Domain and repeats:
15-149
: IPR007921,CHAP domain;
GO term prediction:
MGLPNPKDRKPTASEVVEWALYMAKNRRVIDVDRSYGGQCWDVPNYILERYWGFRTWGNANAMAQKSNYRGRDFKIYRNTASFTPKPGDWAVWANRNPGHVAIVVGPADKNAFVSVDQNWYTANWSGSPPYKIKHTYHDGPGGVTHFVRPPYHPDKTTPAPQPVPK
  BlastP to GMEnzy

Production

  1.  Expressed by pET27a 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 0.023 ± 0.001 ∆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]

Comments

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