BACTERIAL AGGLUTINATION BY A LECTIN FROM THE LEAVES OF THE MEDICINAL PLANT, PIMENTA DIOICA (L.) MERR

Surya P H,Deepti Madayi,Elyas K K

doi:10.22159/ijpps.2018v10i4.20068

Surya P H, Deepti Madayi + Show 1 more

Open Access

https://doi.org/10.22159/ijpps.2018v10i4.20068

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Abstract

Objective: The current investigation involves the purification, characterization of the lectin from the leaves of Pimenta dioica (L.) Merr. (Myrtaceae) a medicinal plant, and its application in bacterial typing.Methods: A lectin was purified from the leaves by cation exchange chromatography. SDS PAGE revealed the molecular weight of the purified lectin. Biochemical characterization was carried out by performing various tests. Hemagglutination inhibition was conducted to detect the sugar specificity. Additionally, bacterial agglutination was performed to predict whether the purified lectin was able to agglutinate the bacterial strains.Results: SDS PAGE analysis revealed the lectin to be a tetramer in the range of 43-66 kDa. The purified lectin agglutinated human, avian, and mouse erythrocytes, and was inhibited by 125 mmol of mannose and xylose. The lectin was stable at 0-60 ° C for 30 min and was unaffected by either 2-Mercaptoethanol (2-ME) or Dithiothreitol (DTT) (50-250µM). A pH of 6.0–8.0 was found optimum for its activity and was nearly independent of metal ions. The purified lectin contained about 20% carbohydrate as estimated by Anthrone method. Purified lectin agglutinated the Gram-negative Escherichia coli and Proteus vulgaris.Conclusion: The isolated lectin was found to possess significant hemagglutinating activity. Due to its ability to agglutinate Gram negative bacteria such as Escherichia coli and Proteus vulgaris, it could be used for bacterial typing and for the design of bacterial filters.

Highlights

Leaves of Pimenta dioica (L.) Merr are found to be a good source of lectin with significant hemagglutination activity
The lectin was purified partially by cation exchange chromatography when glycineNaOH was used as the buffer system and was found to contain nearly 20% carbohydrates
The activity of the lectin did not exhibit any specificity to human, mice or avian erythrocytes and was nearly independent of metal ions

Summary

Methods

A lectin was purified from the leaves by cation exchange chromatography. SDS PAGE revealed the molecular weight of the purified lectin. Bacterial agglutination was performed to predict whether the purified lectin was able to agglutinate the bacterial strains. Bovine Serum Albumin (BSA), periodic acid, and CM-cellulose (Sigma, product code-C0806) was purchased from Sigma-Aldrich (St Louis MO, USA). Amicon Ultra-4 K centrifugal filter (Merck, Product code-UFC801024) was obtained from Merck Millipore Ltd. Blood bags for the purpose of hemagglutination was procured from Malabar Institute of Medical Science (MIMS) Hospital, Calicut. Procurement of plant material and preparation of crude extract. Plants were collected from the Calicut University Botanical Garden, (11ο08′02.01′′N 75ο53′26.28′′E) and a specimen were deposited in the Calicut University Herbarium with a voucher number-6916. Dried leaves were sheared into small pieces

Results

Discussion

Conclusion