Exploring the enzymatic and antibacterial activities of novel mycobacteriophage lysin b enzymes
(2020) In International Journal of Molecular Sciences 21(9).- Abstract
Mycobacteriophages possess different sets of lytic enzymes for disruption of the complex cell envelope of the mycobacteria host cells and release of the viral progeny. Lysin B (LysB) enzymes are mycolylarabinogalactan esterases that cleave the ester bond between the arabinogalactan and mycolic acids in the mycolylarabinogalactan-peptidoglycan (mAGP) complex in the cell envelope of mycobacteria. In the present study, four LysB enzymes were produced recombinantly and characterized with respect to their enzymatic and antibacterial activities. Examination of the kinetic parameters for the hydrolysis of para-nitrophenyl ester substrates, shows LysB-His6 enzymes to be active against a range of substrates (C4-C16), with a catalytic... (More)
Mycobacteriophages possess different sets of lytic enzymes for disruption of the complex cell envelope of the mycobacteria host cells and release of the viral progeny. Lysin B (LysB) enzymes are mycolylarabinogalactan esterases that cleave the ester bond between the arabinogalactan and mycolic acids in the mycolylarabinogalactan-peptidoglycan (mAGP) complex in the cell envelope of mycobacteria. In the present study, four LysB enzymes were produced recombinantly and characterized with respect to their enzymatic and antibacterial activities. Examination of the kinetic parameters for the hydrolysis of para-nitrophenyl ester substrates, shows LysB-His6 enzymes to be active against a range of substrates (C4-C16), with a catalytic preference towards p-nitrophenyl laurate (C12). With p-nitrophenyl butyrate as substrate, LysB-His6 enzymes showed highest activity at 37◦C. LysB-His6 enzymes also hydrolyzed different Tween substrates with highest activity against Tween 20 and 80. Metal ions like Ca2+ and Mn2+ enhanced the enzymatic activity of LysB-His6 enzymes, while transition metal ions like Zn2+ and Cu2+ inhibited the enzymatic activity. The mycolylarabinogalactan esterase activity of LysB-His6 enzymes against mAGP complex was confirmed by LC-MS. LysB-His6 enzymes showed marginal antibacterial activity when tested alone against Mycobacterium smegmatis, however a synergetic activity was noticed when combined with outer membrane permealizers. These results confirm that LysB enzymes are lipolytic enzymes with potential application as antimycobacterials.
(Less)
- author
- Abouhmad, Adel LU ; Korany, Ahmed H. ; Grey, Carl LU ; Dishisha, Tarek LU and Hatti-Kaul, Rajni LU
- organization
- publishing date
- 2020-05-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Antimycobacterial, Lipolytic, LysB, Mycobacteriophage, Mycolylarabinogalactan esterase, Mycolylarabinogalactan-peptidoglycan complex
- in
- International Journal of Molecular Sciences
- volume
- 21
- issue
- 9
- article number
- 3176
- publisher
- MDPI AG
- external identifiers
-
- pmid:32365915
- scopus:85084049229
- ISSN
- 1661-6596
- DOI
- 10.3390/ijms21093176
- language
- English
- LU publication?
- yes
- id
- 4f233f7e-e3ee-4343-bc11-92c12e6a4e5d
- date added to LUP
- 2020-05-09 11:06:06
- date last changed
- 2024-05-01 10:06:09
@article{4f233f7e-e3ee-4343-bc11-92c12e6a4e5d,
abstract = {{<p>Mycobacteriophages possess different sets of lytic enzymes for disruption of the complex cell envelope of the mycobacteria host cells and release of the viral progeny. Lysin B (LysB) enzymes are mycolylarabinogalactan esterases that cleave the ester bond between the arabinogalactan and mycolic acids in the mycolylarabinogalactan-peptidoglycan (mAGP) complex in the cell envelope of mycobacteria. In the present study, four LysB enzymes were produced recombinantly and characterized with respect to their enzymatic and antibacterial activities. Examination of the kinetic parameters for the hydrolysis of para-nitrophenyl ester substrates, shows LysB-His<sub>6</sub> enzymes to be active against a range of substrates (C4-C16), with a catalytic preference towards p-nitrophenyl laurate (C12). With p-nitrophenyl butyrate as substrate, LysB-His<sub>6</sub> enzymes showed highest activity at 37<sup>◦</sup>C. LysB-His<sub>6</sub> enzymes also hydrolyzed different Tween substrates with highest activity against Tween 20 and 80. Metal ions like Ca<sup>2+</sup> and Mn<sup>2+</sup> enhanced the enzymatic activity of LysB-His<sub>6</sub> enzymes, while transition metal ions like Zn<sup>2+</sup> and Cu<sup>2+</sup> inhibited the enzymatic activity. The mycolylarabinogalactan esterase activity of LysB-His<sub>6</sub> enzymes against mAGP complex was confirmed by LC-MS. LysB-His<sub>6</sub> enzymes showed marginal antibacterial activity when tested alone against Mycobacterium smegmatis, however a synergetic activity was noticed when combined with outer membrane permealizers. These results confirm that LysB enzymes are lipolytic enzymes with potential application as antimycobacterials.</p>}},
author = {{Abouhmad, Adel and Korany, Ahmed H. and Grey, Carl and Dishisha, Tarek and Hatti-Kaul, Rajni}},
issn = {{1661-6596}},
keywords = {{Antimycobacterial; Lipolytic; LysB; Mycobacteriophage; Mycolylarabinogalactan esterase; Mycolylarabinogalactan-peptidoglycan complex}},
language = {{eng}},
month = {{05}},
number = {{9}},
publisher = {{MDPI AG}},
series = {{International Journal of Molecular Sciences}},
title = {{Exploring the enzymatic and antibacterial activities of novel mycobacteriophage lysin b enzymes}},
url = {{http://dx.doi.org/10.3390/ijms21093176}},
doi = {{10.3390/ijms21093176}},
volume = {{21}},
year = {{2020}},
}