Title |
Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin |
Author |
Jeonghyun Lim1, Jaeyeon Jang1, Heejoon Myung1,2, and Miryoung Song1* |
Address |
1Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Yongin 17035, Republic of Korea, 2LyseNTech Co., Ltd., Seongnam 13486, Republic of Korea |
Bibliography |
Journal of Microbiology, 60(8),859-866, 2022,
|
DOI |
10.1007/s12275-022-2107-y
|
Key Words |
bacteriophage, endolysin, cell-penetrating peptide,
Acinetobacter baumannii, multiple drug-resistant |
Abstract |
Antimicrobial agents targeting peptidoglycan have shown
successful results in eliminating bacteria with high selective
toxicity. Bacteriophage encoded endolysin as an alternative
antibiotics is a peptidoglycan degrading enzyme with a low
rate of resistance. Here, the engineered endolysin was developed
to defeat multiple drug-resistant (MDR) Acinetobacter
baumannii. First, putative endolysin PA90 was predicted by
genome analysis of isolated Pseudomonas phage PBPA. The
His-tagged PA90 was purified from BL21(DE3) pLysS and
tested for the enzymatic activity using Gram-negative pathogens
known for having a high antibiotic resistance rate including
A. baumannii. Since the measured activity of PA90
was low, probably due to the outer membrane, cell-penetrating
peptide (CPP) DS4.3 was introduced at the N-terminus
of PA90 to aid access to its substrate. This engineered endolysin,
DS-PA90, completely killed A. baumannii at 0.25 μM,
at which concentration PA90 could only eliminate less than
one log in CFU/ml. Additionally, DS-PA90 has tolerance to
NaCl, where the ~50% of activity could be maintained in the
presence of 150 mM NaCl, and stable activity was also observed
with changes in pH or temperature. Even MDR A. baumannii
strains were highly susceptible to DS-PA90 treatment:
five out of nine strains were entirely killed and four strains
were reduced by 3–4 log in CFU/ml. Consequently, DS-PA90
could protect waxworm from A. baumannii-induced death
by ~70% for ATCC 17978 or ~44% for MDR strain 1656-2
infection. Collectively, our data suggest that CPP-fused endolysin
can be an effective antibacterial agent against Gramnegative
pathogens regardless of antibiotics resistance mechanisms. |