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Antibacterial properties of Acinetobacter baumannii phage Abp1 endolysin (PlyAB1).

Huang G, Shen X, Gong Y, Dong Z, Zhao X, Shen W, Wang J, Hu F, Peng Y - BMC Infect. Dis. (2014)

Bottom Line: Acinetobacter baumannii has emerged as one of the most important hospital-acquired pathogens in the world, because of its resistance to almost all available antibiotic drugs.These isolates were shown to belong to different ST clones by multilocus sequence typing.The results presented here show that PlyAB1 has potential as an antibiotic against drug-resistant A. baumannii.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burn Research, Southwest Hospital, Third Military Medical University, Chongqing, China. haitao3140@sina.com.

ABSTRACT

Background: Acinetobacter baumannii has emerged as one of the most important hospital-acquired pathogens in the world, because of its resistance to almost all available antibiotic drugs. Endolysins from phages are attracting increasing interest as potential antimicrobial agents, especially for drug-resistant bacteria. We previously isolated and characterized Abp1, a virulent phage targeting the multidrug-resistant A. baumannii strain, AB1.

Methods: To evaluate the antimicrobial potential of endolysin from the Abp1 phage, the endolysin gene plyAB1 was cloned and over-expressed in Escherichia coli, and the lytic activity of the recombinant protein (PlyAB1) was tested by turbidity assessment and bacteria counting assays.

Results: PlyAB1 exhibits a marked lytic activity against A. baumannii AB1, as shown by a decrease in the number of live bacteria following treatment with the enzyme. Moreover, PlyAB1 displayed a highly specific lytic effect against all of the 48 hospital-derived pandrug-resistant A. baumannii isolates that were tested. These isolates were shown to belong to different ST clones by multilocus sequence typing.

Conclusions: The results presented here show that PlyAB1 has potential as an antibiotic against drug-resistant A. baumannii.

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Related in: MedlinePlus

Lytic activity of PlyAB1 against the 48 clinical PDRAB isolates. (A) PlyAB1 lytic activity results for 48 PDRAB isolates 30 min after addition of the enzyme. Blue bar denotes the OD600 at 0 min, red bar denotes the OD600 30 min later. (B) Rows A, C, E, and G represent the negative control groups containing buffer lacking PlyAB1 protein. Rows B, D, F, and H represent the trial groups (with PlyAB1 protein). Changes in the OD600 at 30 min were recorded by a microplate reader. (C and D) Lytic range of PlyAB1. Left to right: AB1, BL21, JM109, N315, and PAO1. PlyAB1 can only lyse A. baumannii AB1.
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Fig5: Lytic activity of PlyAB1 against the 48 clinical PDRAB isolates. (A) PlyAB1 lytic activity results for 48 PDRAB isolates 30 min after addition of the enzyme. Blue bar denotes the OD600 at 0 min, red bar denotes the OD600 30 min later. (B) Rows A, C, E, and G represent the negative control groups containing buffer lacking PlyAB1 protein. Rows B, D, F, and H represent the trial groups (with PlyAB1 protein). Changes in the OD600 at 30 min were recorded by a microplate reader. (C and D) Lytic range of PlyAB1. Left to right: AB1, BL21, JM109, N315, and PAO1. PlyAB1 can only lyse A. baumannii AB1.

Mentions: In the lytic activity assay, PlyAB1 digested all 48 of the hospital-derived PDRAB isolates (Figure 5A, B). As shown in Figure 5A, all the OD600 values of the PDRAB isolates and PlyAB1 decreased after 30 min incubation. The OD600 of the PDRAB isolates dropped significantly when purified PlyAB1 was incubated with them (Figure 5B, rows B, D, F, and H). For the negative control, the cell densities of the PDRAB isolates incubated with the same volume of reaction buffer as was used for the test samples did not change significantly (Figure 5B, row A, C, E, and G). In an even broader test, we found that PlyAB1 could rapidly lyse 206 of 212 additional clinical MDRAB isolates in 30 minutes (see Additional files 2 and 3).Figure 5


Antibacterial properties of Acinetobacter baumannii phage Abp1 endolysin (PlyAB1).

Huang G, Shen X, Gong Y, Dong Z, Zhao X, Shen W, Wang J, Hu F, Peng Y - BMC Infect. Dis. (2014)

Lytic activity of PlyAB1 against the 48 clinical PDRAB isolates. (A) PlyAB1 lytic activity results for 48 PDRAB isolates 30 min after addition of the enzyme. Blue bar denotes the OD600 at 0 min, red bar denotes the OD600 30 min later. (B) Rows A, C, E, and G represent the negative control groups containing buffer lacking PlyAB1 protein. Rows B, D, F, and H represent the trial groups (with PlyAB1 protein). Changes in the OD600 at 30 min were recorded by a microplate reader. (C and D) Lytic range of PlyAB1. Left to right: AB1, BL21, JM109, N315, and PAO1. PlyAB1 can only lyse A. baumannii AB1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4274762&req=5

Fig5: Lytic activity of PlyAB1 against the 48 clinical PDRAB isolates. (A) PlyAB1 lytic activity results for 48 PDRAB isolates 30 min after addition of the enzyme. Blue bar denotes the OD600 at 0 min, red bar denotes the OD600 30 min later. (B) Rows A, C, E, and G represent the negative control groups containing buffer lacking PlyAB1 protein. Rows B, D, F, and H represent the trial groups (with PlyAB1 protein). Changes in the OD600 at 30 min were recorded by a microplate reader. (C and D) Lytic range of PlyAB1. Left to right: AB1, BL21, JM109, N315, and PAO1. PlyAB1 can only lyse A. baumannii AB1.
Mentions: In the lytic activity assay, PlyAB1 digested all 48 of the hospital-derived PDRAB isolates (Figure 5A, B). As shown in Figure 5A, all the OD600 values of the PDRAB isolates and PlyAB1 decreased after 30 min incubation. The OD600 of the PDRAB isolates dropped significantly when purified PlyAB1 was incubated with them (Figure 5B, rows B, D, F, and H). For the negative control, the cell densities of the PDRAB isolates incubated with the same volume of reaction buffer as was used for the test samples did not change significantly (Figure 5B, row A, C, E, and G). In an even broader test, we found that PlyAB1 could rapidly lyse 206 of 212 additional clinical MDRAB isolates in 30 minutes (see Additional files 2 and 3).Figure 5

Bottom Line: Acinetobacter baumannii has emerged as one of the most important hospital-acquired pathogens in the world, because of its resistance to almost all available antibiotic drugs.These isolates were shown to belong to different ST clones by multilocus sequence typing.The results presented here show that PlyAB1 has potential as an antibiotic against drug-resistant A. baumannii.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burn Research, Southwest Hospital, Third Military Medical University, Chongqing, China. haitao3140@sina.com.

ABSTRACT

Background: Acinetobacter baumannii has emerged as one of the most important hospital-acquired pathogens in the world, because of its resistance to almost all available antibiotic drugs. Endolysins from phages are attracting increasing interest as potential antimicrobial agents, especially for drug-resistant bacteria. We previously isolated and characterized Abp1, a virulent phage targeting the multidrug-resistant A. baumannii strain, AB1.

Methods: To evaluate the antimicrobial potential of endolysin from the Abp1 phage, the endolysin gene plyAB1 was cloned and over-expressed in Escherichia coli, and the lytic activity of the recombinant protein (PlyAB1) was tested by turbidity assessment and bacteria counting assays.

Results: PlyAB1 exhibits a marked lytic activity against A. baumannii AB1, as shown by a decrease in the number of live bacteria following treatment with the enzyme. Moreover, PlyAB1 displayed a highly specific lytic effect against all of the 48 hospital-derived pandrug-resistant A. baumannii isolates that were tested. These isolates were shown to belong to different ST clones by multilocus sequence typing.

Conclusions: The results presented here show that PlyAB1 has potential as an antibiotic against drug-resistant A. baumannii.

Show MeSH
Related in: MedlinePlus