Detail of GM0157


Summary

The genetically modified enzybiotic, named Mdc-hly , constructed by Fusion to Peptides strategy, change or extend lytic spectrum.
With  Mdc-hly
  Sequence Length:  178 AA.
  Mass:  19032.3 Da.
  Isoelectric Point:  9.74
  Function:  The recombinant fusion protein Mdc–hly has improved in vitro antimicrobial activity and action spectrum compared to Mdc and hly.

Construction

GM0157, constructed by Fusion to Peptides strategy.
Q95VE8: Cecropin
P61626: Lysozyme C
UniProt

PF00272?:28-60
IPR023346:19-147
InterPro

GM0157:1-33
GM0157:49-178
GMEnzy

30
60
63
30
60
90
120
148

Details:
GM0157[1-33]: derive from protein Q95VE8 sequence 28-60

Q95VE8, 63 AA., the Cecropin from Musca domestica (House fly)

Source: Musca domestica (House fly)

Domains and repeats

28-60:  PF00272?, the Cecropin

GO term prediction

Cellular Component:  0005576, the extracellular region?


Linking to UniprotKB
Linging to InterPro
GM0157[49-178]: derive from protein P61626 sequence 19-148

P61626, 148 AA., the Lysozyme C from Homo sapiens (Human)?

Source: Homo sapiens (Human)?

Domains and repeats

19-147:  IPR023346, the Lysozyme-like domain

GO term prediction

Biological Process:  0016998, the cell wall macromolecule catabolic process?
Molecular Function:  0003796, the lysozyme activity?


Linking to UniprotKB
Linging to InterPro

Annotation

1-33
49-178
50
100
150
200
211

1-33
: PEP, cecropin,cecropin;
49-178
: EAD, Lysozyme domain, derived from Human lysozyme;
Domain and repeats:
1-33
: PF00272?,Cecropin;
49-177
: IPR023346,Lysozyme-like domain;
GO term prediction:
KIGKKIERVQQHTRDATIQTIGVAGGAANVAATGGGGSGGGGSGGGGSKVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV
  BlastP to GMEnzy

Production

  1.  Expressed by pET-32a in E. coli BL21 (DE3),  Purified by immobilized-metal affinity chromatography according to the instruction of HisTrap kit (GE healthcare

Activity

  1.  MIC test on Staphylococcus aureus (Staphylococcus aureus(S. aureus) ATCC25923) by the turbidity-reduction assays
  showed 3.12μM
  2.  MIC test on Staphylococcus epidermidis (Staphylococcus epidermidis(S. epidermidis) ATCC 12) by the turbidity-reduction assays
  showed 6.25μM
  3.  MIC test on Clostridium sporogenes (Clostridium sporogenes (C. sporogenes) ATCC 11437) by the turbidity-reduction assays
  showed 1.56μM
  4.  MIC test on Micrococcus luteus (Micrococcus lysodeikticus(M. lysodeikticus) CICC 2) by the turbidity-reduction assays
  showed 3.12μM
  5.  MIC test on Escherichia coli (Escherichia coli (E. coli) ATCC25922") by the turbidity-reduction assays
  showed 0.39μM
  6.  MIC test on Shigella flexneri (Shigella flexneri (S. flexneri) ATCC 12022) by the turbidity-reduction assays
  showed 0.78μM
  7.  MIC test on Pseudomonas aeruginosa (Pseudomonas aeruginosa (P. aeruginosa)ATCC27853) by the turbidity-reduction assays
  showed 6.25μM
  8.  MIC test on Salmonella paratyphi-B (Salmonella paratyphi-B (S. paratyphi-B) CICC 2149) by the turbidity-reduction assays
  showed 1.56μM
  9.  MIC test on Enterobacter cloacae (Enterobacter cloacae (E. cloacae) CICC 21652) by the turbidity-reduction assays
  showed 1.56μM
  10.  MBC test on Staphylococcus aureus (Staphylococcus aureus(S. aureus) ATCC25923) by the turbidity-reduction assays
  showed 6.25μM
  11.  MBC test on Staphylococcus epidermidis (Staphylococcus epidermidis(S. epidermidis) ATCC 12) by the turbidity-reduction assays
  showed 12.5μM
  12.  MBC test on Clostridium sporogenes (Clostridium sporogenes (C. sporogenes) ATCC 11437) by the turbidity-reduction assays
  showed 3.12μM
  13.  MBC test on Micrococcus luteus (Micrococcus lysodeikticus(M. lysodeikticus) CICC 2) by the turbidity-reduction assays
  showed 6.25μM
  14.  MBC test on Escherichia coli (Escherichia coli (E. coli) ATCC25922) by the turbidity-reduction assays
  showed 0.39μM
  15.  MBC test on Shigella flexneri (Shigella flexneri (S. flexneri) ATCC 12022) by the turbidity-reduction assays
  showed 1.56μM
  16.  MBC test on Pseudomonas aeruginosa (Pseudomonas aeruginosa (P. aeruginosa)ATCC27853) by the turbidity-reduction assays
  showed 12.5μM
  17.  MBC test on Salmonella paratyphi-B (Salmonella paratyphi-B (S. paratyphi-B) CICC 2149) by the turbidity-reduction assays
  showed 1.56μM
  18.  MBC test on Enterobacter cloacae (Enterobacter cloacae (E. cloacae) CICC 21652) by the turbidity-reduction assays
  showed 3.12μM

Reference

  1. Lu, X. M. Jin, X. B. Zhu, J. Y. Mei, H. F. Ma, Y. Chu, F. J. Wang, Y. Li, X. B.. (2010) Expression of the antimicrobial peptide cecropin fused with human lysozyme in Escherichia coli. Appl Microbiol Biotechnol. 876:2169-2176. [doi:10.1007/s00253-010-2606-3.] [PMID:20499232]

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