Lysostaphin: Engineering and Potentiation toward Better Applications

Abstract

Lysostaphin is a potent bacteriolytic enzyme with endopeptidase activity against the common pathogen Staphylococcus aureus. By digesting the pentaglycine crossbridge in the cell wall peptidoglycan of S. aureus including the methicillin-resistant strains, lysostaphin initiates rapid lysis of planktonic and sessile cells (biofilms) and has great potential for use in agriculture, food industries, and pharmaceutical industries. In the past few decades, there have been tremendous efforts in potentiating lysostaphin for better applications in these fields, including engineering of the enzyme for higher potency and lower immunogenicity with longer-lasting effects, formulation and immobilization of the enzyme for higher stability and better durability, and recombinant expression for low-cost industrial production and in situ biocontrol. These achievements are extensively reviewed in this article focusing on applications in disease control, food preservation, surface decontamination, and pathogen detection. In addition, some basic properties of lysostaphin that have been controversial and only elucidated recently are summarized, including the substrate-binding properties, the number of zinc-binding sites, the substrate range, and the cleavage site in the pentaglycine crossbridge. Resistance to lysostaphin is also highlighted with a focus on various mechanisms. This article is concluded with a discussion on the limitations and future perspectives for the actual applications of lysostaphin.

Peptidoglycan-Targeting Staphylolytic Enzyme Lysostaphin as a Novel and Efficient Protease toward Glycine-Rich Flexible Peptide Linkers

“…Additionally, compared with TEV protease that is confronted with low yield, poor solubility, and instability during recombinant expression, lysostaphin can be expressed with high yields and stability intracellularly or extracellularly in various hosts including E. coli, Bacillus subtilis, Lactococcus lactis, Pichia pastoris, and mammalian cells as long as the codons are optimized according to the preference of the hosts and the glycosylation sites are mutated (for eukaryotic systems). ,,,− This feature greatly facilitates lysostaphin production at both the lab scale and the industrial scale. However, it is worth noting that lysostaphin as a protease has its intrinsic limitations.…”

“…Lysostaphin is an endopeptidase naturally secreted by Staphylococcus simulans biovar staphylolyticus against the competing bacterium Staphylococcus aureus (S. aureus), and it cleaves the cell wall peptidoglycan of S. aureus via specific digestion of the pentaglycine crossbridge. 6 Besides the natural cell wall of S. aureus, lysostaphin is also able to digest synthetic peptides such as N-acetylhexaglycine, 7 oligoglycine containing two to six glycine residues, 8 and KGGGGG/AEKAGGGGG peptides with different fluorescent tags. 9,10 Surprisingly, in a study of identifying elastin-binding proteins in S. aureus using an elastin column, it was found that lysostaphin used to solubilize the cells was bound to the column and could digest elastin, which is an insoluble protein with one-third of its amino acids being glycine.…”

“…aureus), and it cleaves the cell wall peptidoglycan of S. aureus via specific digestion of the pentaglycine crossbridge . Besides the natural cell wall of S.…”