PMID:17592147

Citation	Lu, TK and Collins, JJ (2007) Dispersing biofilms with engineered enzymatic bacteriophage. Proc. Natl. Acad. Sci. U.S.A. 104:11197-202
Abstract	Synthetic biology involves the engineering of biological organisms by using modular and generalizable designs with the ultimate goal of developing useful solutions to real-world problems. One such problem involves bacterial biofilms, which are crucial in the pathogenesis of many clinically important infections and are difficult to eradicate because they exhibit resistance to antimicrobial treatments and removal by host immune systems. To address this issue, we engineered bacteriophage to express a biofilm-degrading enzyme during infection to simultaneously attack the bacterial cells in the biofilm and the biofilm matrix, which is composed of extracellular polymeric substances. We show that the efficacy of biofilm removal by this two-pronged enzymatic bacteriophage strategy is significantly greater than that of nonenzymatic bacteriophage treatment. Our engineered enzymatic phage substantially reduced bacterial biofilm cell counts by approximately 4.5 orders of magnitude ( approximately 99.997% removal), which was about two orders of magnitude better than that of nonenzymatic phage. This work demonstrates the feasibility and benefits of using engineered enzymatic bacteriophage to reduce bacterial biofilms and the applicability of synthetic biology to an important medical and industrial problem.
Links	PubMed PMC1899193 Online version:10.1073/pnas.0704624104
Keywords	Bacteriophage T3/enzymology; Bacteriophage T3/genetics; Bacteriophage T7/enzymology; Bacteriophage T7/genetics; Biofilms/growth & development; Escherichia coli/genetics; Escherichia coli/physiology; Escherichia coli/virology; Extracellular Matrix/enzymology; Extracellular Matrix/genetics; Extracellular Matrix/virology; Genetic Engineering/methods
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Significance

Shedding light on the use of synthetic biology where phage could be engineered to deal with real world problems such as eradication of biofilm or perhaps the fight against antibiotic resistant microbes. This paper illustrates the efficacy of phages that are engineered to produce the enzyme necessary for the degradation of polysaccharide responsible for biofilm formation. Doing so allows phage to diffuse easily and adsorb to next host for killing. The data presented in this papers has been able to reflect some aspect of it, however, few comments have been gathered from the valuable readers:

I) What was the necessity for using 10B-S.tag in the control is not clearly described. Isogenic control would be without the S.tag as well.

II) T3wt is almost equally efficient compared to the engineered T7DspB phage that can now avoid exclusion due to F-plasmid. Isn't isolating phage with classical genetic approach that resist the exclusion a better option?

III) If the viable cell count after biofilm treatment with engineered phage still persist to some level(Fig.3B), how can we rule out the possibility that phage replication has in fact occurred within the planktonic cells that are on the surface of the biofilm?

IV)Wouldn't the 20 hour untreated biofilm SEM image look more dense (Fig. 4C)?

V) Each data points for Fig.4 E&F represents different set of experiment with respect to initial phage inoculation. How about illustrating this dose response in a bar diagram instead of connecting two separate events?

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