Detail of GM0013
Summary
The genetically modified enzybiotic, named Ply187AN-KSH3b , constructed by Domains Assembly strategy, improve the lytic activity.With Ply187AN-KSH3b
Sequence Length: 225 AA.
Mass: 25079.1 Da.
Isoelectric Point: 9.33
Function: The fusion’s MIC is five- fold lower than that of LysK’s (and twofold lower when compared on a molar basis)
Construction
GM0013, constructed by Domains Assembly strategy. No schema construted on GM0013
Details:
GM0013[1-157]: derive from protein O56785 sequence 1-157
O56785, 628 AA., the Cell wall hydrolase Ply187 from Staphylococcus phage 187
Source: Staphylococcus phage 187
476-622: IPR013338, the Lysozyme domain, subfamily 2
14-145: IPR007921, the CHAP domain
476-622: 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?
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
Domains and repeats
11-145: IPR007921, the CHAP domain476-622: IPR013338, the Lysozyme domain, subfamily 2
14-145: IPR007921, the CHAP domain
476-622: 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?
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
GM0013[158-293]: derive from protein I6X5A8 sequence 409-481
I6X5A8, 495 AA., the LysK from Staphylococcus phage Fi200W
Source: Staphylococcus phage Fi200W
196-369: IPR002502, the N-acetylmuramoyl-L-alanine amidase domain
409-481: IPR003646, the SH3-like domain, bacterial-type?
Linking to UniprotKB
Linging to InterPro
Domains and repeats
29-160: IPR007921, the CHAP domain196-369: IPR002502, the N-acetylmuramoyl-L-alanine amidase domain
409-481: IPR003646, the SH3-like domain, bacterial-type?
GO term prediction
Molecular Function: 0008745, the N-acetylmuramoyl-L-alanine amidase activityLinking to UniprotKB
Linging to InterPro
Annotation
1-157
158-293
1-157
: EAD, CHAP domain, derived from S. aureus bacteriophage 187 endolysin;158-293
: CBD, SH3b domain, derived from anti-staphylococcal bacteriophage endolysin, LysK;Domain and repeats:
11-145
: IPR007921,CHAP domain; 14-145
: IPR007921,CHAP domain; 158-293
: IPR003646,SH3-like domain, bacterial-type?; GO term prediction:
MALPKTGKPTAKQVVDWAINLIGSGVDVDGYYGRQCWDLPNYIFNRYWNFKTPGNARDMAWYRYPEGFKVFRNTSDFVPKPGDIAVWTGGNYNWNTWGHTGIVVGPSTKSYFYSVDQNWNNSNSYVGSPAAKIKHSYFGVTHFVRPAYKAEPKPTP-ENATFVNGNQPIVTRIGSPFLNAPVGGNLPAGATIVYDEVCIQAGHIWIGYNAYNGNRVYCPVRTCQGV
Production
1. Expressed by pET21a in E. coli BL21(DE3), Purified by nickel chromatography Ni-NTA (Qiagen, Valencia, CA)Activity
1. MIC test on Staphylococcus aureus (S. aureus Newman) by microdilution broth methodshowed 7.6±2.9μg/mL(0.24±0.09 nmol/mL)