Design and Stabilization of Natural Antibacterial Compound Allicin against Methicillin-Resistant Staphylococcus Aureus for Treatment as a Novel Antibiotic

 

Bezlon G., Shanmugha Sundhar D., Rinu Edwin R.E.

Udaya School of Engineering, Udaya Nagar, Nagercoil, Kanyakumari district.

 

ABSTRACT:

Antibiotic resistance is the biggest threat facing human health nowadays. Control of the spread of antibiotic-resistant bacteria and the treatment of infections caused by them is a major problem Worldwide. In particular, methicillin -resistant Staphylococcus aureus (MRSA) presents major infection control problems. It is estimated that the frequency of infection cases occuring in hospitals due to methicillin-resistant Staphylococcus aureus has doubled over the past five years. In concern with the infection, patients are treated with antibiotics such as vancomycin, that has higher possibility to side-effects such as nephrotoxicity. A natural pigment, Allicin, often referred to as the natural antibiotic, extracted from garlic and ginger has proved to fight against mrsa (methicillin -resistant Staphylococcus aureus) and a wide range of bacterial species and fungal infections.  Unlike other antibiotics, its S(=O)S thiosulphinate group in allicin is said to react with a variety of SH containing enzymes within the bacterial cell. It is reported to inhibit the acetyl CoA forming system, to inhibit DNA and protein synthesis. This compound however is highly volatile and unstable. The outlook of our work is to provide a novel antibiotic for the treatment of Methicillin resistant Staphyllococcus aureus and other multi drug resistant, disease causing bacteria using Allicin. A few methods of stabilization of allicin were undertaken and the methods of supplement and the results were discussed. In this study aqueous extract of allicin and chemically treated crystalline Allicin (extracted from garlic) were tested on clinical isolates of MRSA that survived methicillin and the results that were compared statistically and graphically are provided.

 

INTRODUCTION:

MRSA:

Antibiotic resistance is referred as the drug resistance a microorganism acquires to withstand the effect of antibiotics. Recent studies suggest that most of the bacteria are resistant to various antibiotics. The resistance is achieved by various mechanisms like altering the target site or by inactivating the drug by releasing certain enzymes. This paper focusses on the antibiotic resistance of Staphylococcus aureus and its treatment using Allicin. Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterium responsible for several infections in humans that are difficult to treat.

 

It is also called oxacillin-resistant Staphylococcus aureus (ORSA). MRSA is any strain of  that has developed, through the process of natural selection, resistance to beta-lactam antibiotics, like the penicillins, methicillin, dicloxacillin, nafcilin, oxacilin etc.The resistance does make MRSA infection more difficult to treat with standard types of antibiotics and thus more dangerous. The increasing prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals and the community has led to a demand for new agents that could be used to decrease the spread of these bacteria.

 

Allicin against MRSA:

Allicin is an organo sulphur compound, referred to as the natural antibiotic, extracted from garlic has proven to fight against mrsa and a wide range of bacterial species and fungal infections.

 

Figure-1

Sourcehttps://patentimages.storage.googleapis.com/US20070036875A1/US20070036875A1-20070215-C00002.png

 

This compound also possesses anti-fungal properties. It has the thiosulfinate functional group, R-S(O)-S-R. The compound is not present in garlic unless tissue damage occurs, and is formed by the action of the enzyme allinase on allin.

 

(SCH₂CH=CH₂)₂ + RCO₃H →

(Allin)

CH₂=CHCH₂S (O)SCH₂CH=CH₂ + RCO₂H

  (Allicin)

 

Allicin has been found to have numerous antimicrobial properties, One potential application is in the treatment of  methicillin -resistant Staphylococcus aureus (MRSA). Allicin can restore the resistance of many dangerous bacterial strains. It can break up a biofilm, destroy a wide range of bacterial species, wipe out fungal infections, boost an under active immune system, reduce cholesterol and blood pressure levels, prevent viral infections, kill off parasites, remove protozoal organisms, vasodilate when necessary, prevent the release of histamine, and even prevent mosquitoes from attacking. Low concentrations of allicin can destroy all bacterial cells.

 

The Allicin molecule is small and easily penetrates cell walls. This compound is highly active but generally unstable. Alliinase is irreversibly deactivated below pH 3; as such, allicin is generally not produced in the body from the consumption of fresh or powdered garlic. Furthermore, allicin can be unstable, breaking down within 16 h at 23 °C.

 

 

Figure2Source:https://patentimages.storage.googleapis.com/US20070036875A1/US20070036875A1-20070215-C00002.png

 

MATERIALS AND METHODS:

Several methods can be employed to stabilize allicin. A water-based formulation of purified allicin was found to be more chemically stable than other preparations of garlic extracts. Crystalline products of Allicin however showed relatively low biological activity. Enzyme immobilized Allicin prepared by passing the precursors through columns steadily was less effective than aqueous solution of allicin. the stability may be due to the hydrogen bonding of water to the reactive oxygen atom in allicin.

 

Ten strains of Staphyllococcus aureus were cultured in standard laboratory conditions. Strains were cultured overnight at 37°C in agar plates. Methicillin disc of low dosage (20-1000 ug/ml) was placed in the plate and cultured for 24 hours. The colonies that survived in the zone of inhibition were isolated and transferred to another plate. Of the isolates, 2 strains were identified as fully susceptible (zone sizes 33–45 mm), 3 showed intermediate susceptibility (zone sizes 12–23 mm) and 6 strains were resistant (no zone of inhibition). Variability of zone size was considerable The methicillin resistant Staphylococcus aureus were thus cultured for 48 hours. Allicin in various forms were added to the strains and their relative biological activities were tested.

 

 

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Image-1

 

Curcumin-Allicin complex showed drastic antibacterial effects against mrsa, but the implementation of which becomes difficult while treatment in a complex state.

 

Table-1

Concentration of aqueous Allicin solution (ug/ml)	Time period of bacterial inhibition observed in the zone  (days)
250	7
375	10
500	14

 

 

Table -2

           

RESULTS AND CONCLUSIONS:

It was found that allicin was most active against MRSA at concentrations of 125 to 250 ug/ml. No activity was found below 60 ug/ml. Most of the isolates were killed at 250 ul/mg. Under normal concentrations, Alicin killed almost all the strains of mrsa. Even little higher concentrations will not lead to any tissue damage or other side effects. No microorganism is said to have developed resistance in any form against the natural antibiotic Allicin. Thus, Allicin supplied in preferred forms stands to prove as a novel antibiotic.

 

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Received on 27.08.2013                                  Accepted on 01.09.2013        

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