Literature summary for 3.5.1.28 extracted from

Farkasovska, J.; Godany, A.
Characterization of the N-terminal catalytic domain of Lytmu1/6, an endolysin from Streptomyces aureofaciens Phage mu1/6 (2016), Curr. Microbiol., 73, 602-610 .

Cloned(Commentary)

Cloned (Comment)	Organism
sequence comparisons, recombinant expression of His-tagged wild-type and mutant endolysins in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain XL10 GOLD	Streptomyces phage mu1/6

Protein Variants

Protein Variants	Comment	Organism
C116A	site-directed mutagenesis, the mutant shows unaltered activity compared to wild-type	Streptomyces phage mu1/6
C131A	site-directed mutagenesis, the mutant shows unaltered activity compared to wild-type	Streptomyces phage mu1/6
D128A	site-directed mutagenesis, the mutant shows unaltered activity compared to wild-type	Streptomyces phage mu1/6
D148A	site-directed mutagenesis, the mutant shows unaltered activity compared to wild-type	Streptomyces phage mu1/6
D186A	site-directed mutagenesis, inactive mutant	Streptomyces phage mu1/6
D186E	site-directed mutagenesis, the mutant shows moderately reduced activity compared to wild-type	Streptomyces phage mu1/6
D21A	site-directed mutagenesis, the mutant shows slightly reduced activity compared to wild-type	Streptomyces phage mu1/6
D35A	site-directed mutagenesis, the mutant shows moderately reduced activity compared to wild-type	Streptomyces phage mu1/6
D70A	site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type	Streptomyces phage mu1/6
E109A	site-directed mutagenesis, inactive mutant	Streptomyces phage mu1/6
E109D	site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type	Streptomyces phage mu1/6
E181A	site-directed mutagenesis, the mutant shows moderately reduced activity compared to wild-type	Streptomyces phage mu1/6
H176A	site-directed mutagenesis, inactive mutant	Streptomyces phage mu1/6
H184A	site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type	Streptomyces phage mu1/6
H31A	site-directed mutagenesis, inactive mutant	Streptomyces phage mu1/6
H66A	site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type	Streptomyces phage mu1/6
K27A	site-directed mutagenesis, inactive mutant	Streptomyces phage mu1/6
K27R	site-directed mutagenesis, the mutant shows unaltered activity compared to wild-type	Streptomyces phage mu1/6
K87A	site-directed mutagenesis, the mutant shows slightly reduced activity compared to wild-type	Streptomyces phage mu1/6
L52P	site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type	Streptomyces phage mu1/6
additional information	construction of diverse truncation mutants of endolysin. The mutants lacking more than 200 C-terminal amino acids or the cell wall-binding domain are inactive, while N-terminal truncation mutant, or C-terminal truncation mutants lacking less than 200 amino acids still show lytic activity, although less than the wild-type enzyme	Streptomyces phage mu1/6
N99A	site-directed mutagenesis, the mutant shows slightly reduced activity compared to wild-type	Streptomyces phage mu1/6
P71A	site-directed mutagenesis, the mutant shows unaltered activity compared to wild-type	Streptomyces phage mu1/6
Q107A	site-directed mutagenesis, the mutant shows slightly reduced activity compared to wild-type	Streptomyces phage mu1/6
Q78A	site-directed mutagenesis, the mutant shows slightly reduced activity compared to wild-type	Streptomyces phage mu1/6
R100A	site-directed mutagenesis, the mutant shows slightly reduced activity compared to wild-type	Streptomyces phage mu1/6
R97A	site-directed mutagenesis, the mutant shows moderately reduced activity compared to wild-type	Streptomyces phage mu1/6
T33A	site-directed mutagenesis, the mutant shows moderately reduced activity compared to wild-type	Streptomyces phage mu1/6
W140A	site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type	Streptomyces phage mu1/6
W167A	site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type	Streptomyces phage mu1/6
W5A	site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type	Streptomyces phage mu1/6

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cell wall	-	Streptomyces phage mu1/6	5618	-

Organism

Organism	UniProt	Comment	Textmining
Streptomyces phage mu1/6	Q7Y4H8	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant endolysins from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration	Streptomyces phage mu1/6

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	N-acetylmuramoyl-L-alanine amidase activity is measured according to the protocol described previously. This method is based on the degradation of free muramic acid resulting from cleavage of the amide bond in the peptidoglycan to lactic acid followed by degradation of the latter to acetaldehyde which can be determined colorimetrically with 4-phenylphenol. Spectrophotometric measurements	Streptomyces phage mu1/6	?	-	?

Synonyms

Synonyms	Comment	Organism
endolysin	-	Streptomyces phage mu1/6
Lytmu1/6	-	Streptomyces phage mu1/6

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	in vivo assay at	Streptomyces phage mu1/6

General Information

General Information	Comment	Organism
malfunction	various C-terminally truncated versions of Lytmu1/6 fail to cause lysis, indicating the necessity of the cell wall binding domain (CBD) for full enzyme activity	Streptomyces phage mu1/6
additional information	Lytmu1/6 is neither a muramidase nor a glucosaminidase. The endolysin releases significant amounts of free amino acids from the peptidoglycan. The N-terminal domain is responsible for the catalytic activity of Lytmu1/6. Amino acid residues K27, H31, E109, H176, and D186 are important for catalytic activity, mutational analysis	Streptomyces phage mu1/6