Abstract
BackgroundIn unfavourable environment, such as nutrient limitation, some bacteria encased themselves into a three dimensional polymer matrix called biofilm. The majority of microbial infections in human are biofilm related, including chronic lung, wound, and ear infections. The matrix of biofilm which consists of extracellular polymeric substances (EPS) causes bacterial colonization on medical implanted device in patients, such as catheter and lead to patient’s death. Biofilm infections are harder to treat due to increasing antibiotic resistance compared to planktonic microbial cells and escalating the antibiotic concentration may result into in vivo toxicity for the patients. Special compounds which are non-microbicidal that could inhibit or destroy biofilm formation are called antibiofilm compounds, for example enzymes, anti-quorum sensing, and anti-adhesins. Arthrobacter sp. CW01 produced antibiofilm compound known as amylase. This time our preliminary study proved that the antibiofilm compound was not only amylase, but also protease. Therefore, this research aimed to optimize the production of antibiofilm agents using amylase and protease inducing media. The five types of production media used in this research were brain heart infusion (BHI) (Oxoid), BHI with starch (BHIS), casein with starch (CS), yeast extract with starch (YS), and casein-yeast extract with starch (CYS). Biofilm eradication and inhibition activities were assayed against Pseudomonas aeruginosa (ATCC 27,853) and Staphylococcus aureus (ATCC 25,923).ResultsThe results showed that different production media influenced the antibiofilm activity. Addition of starch, casein and yeast extract increased the production of amylase and protease significantly. Higher amylase activity would gradually increase the antibiofilm activity until it reached the certain optimum point. It was shown that crude extracts which contained amylase only (BHI, BHIS and YS) had the optimum eradication activity against P. aeruginosa and S. aureus biofilm around 60–70 %. Meanwhile, CS and CYS crude extracts which contained both amylase and protease increased the biofilm eradication activity against both pathogens, which were around 70–90 %.ConclusionsIt was concluded that the combination of amylase and protease was more effective as antibiofilm agents against P. aeruginosa and S. aureus rather than amylase only.
Highlights
In unfavourable environment, such as nutrient limitation, some bacteria encased themselves into a three dimensional polymer matrix called biofilm
Determination of amylase activity The highest amylase enzyme activity of the crude extract was from BHI with starch (BHIS) at 0.39 IU/mL and the highest specific activity was casein-yeast extract with starch (CYS) crude extract at 2.88 IU/mg (Table 2)
It was shown that crude extracts which contained amylase only (BHI, BHIS and yeast extract with starch (YS)) had maximum eradication activity against P. aeruginosa biofilm around 60–70 %
Summary
In unfavourable environment, such as nutrient limitation, some bacteria encased themselves into a three dimensional polymer matrix called biofilm. The bacteria would attach into a surface and encased themselves in a three dimensional polymer matrix called biofilm as a mechanism to survive harsh living environment. They can switch between planktonic form and biofilm form. Biofilm formation causes bacterial colonization on medical implanted device in patients, such as catheter and lead to the patient’s death. These kinds of infections are hard to diagnose and treat because of increasing antibiotic resistance in biofilm compared to planktonic cells [1,2,3]
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