Malaysian Journal of Analytical Sciences
Vol 19 No 4 (2015): 831 - 840
BACTERIOCIN ISOLATED FROM HALOMONAS
SP.: A BACTERIAL DING PROTEIN?
(Pemencilan Bakteriosin dari Halomonas
sp. : Sejenis Bakteria DING protin?)
Atirah Azemin1*, Peter
Klappa2, Mohd Shahir Shamsir Omar1
1Faculty of Biosciences and
Medical Engineering,
Universiti Teknologi Malaysia,81310 Skudai, Johor,
Malaysia
2School of Biosciences,
Stacey Building,
University of Kent, Canterbury, Kent, CT2 7NJ,United
Kingdom
*Corresponding author: atirahasyifa@yahoo.com
Received:
23 November 2014; Accepted: 27 June 2015
Abstract
A marine halophile, Halomonas sp. strain M3 was
isolated from Straits of Johor, Malaysia and produce bacteriocin CC that acts
as bacteriostatic agent. Characterisation of the bacterium showed that optimal
growth and bacteriocin production is at ambient temperature, pH of 8-8.5 in nutrient
broth medium supplemented with 2.9% w/v NaCI to mimic saltwater conditions.
The stability studies indicated that bacteriocin CC is heat-labile (35°C-50°C)
and was stable over 2 years when stored in 0.02M Tris-HCI with 30-60% glycerol
at 4°C. A loss of activity was detected after proteolytic enzymes treatment,
indicating the proteinaceous nature of the antimicrobial compound. The amino
acid sequence of bacteriocin CC was obtained by Edman degradation and MALDI-TOF
analysis, showed the characteristic sequence of a DING protein
(D-I-N-G-G-G-A-T-L-P-Q-A-L-Y-Q) in size 38.9-kDa at pI of 6.8. These proteins constitute a
conserved and widely distributed set of proteins found in all kingdoms with
ligand-binding activities and hydrolytic enzyme, suggesting a possible role in
cell signalling and bio mineralization in DING isolates. Intriguingly, DING
proteins also have been involved as an anti-tumour agent in humans. Thus,
bacteriocin CC as DING protein family members should be further studied to
investigate its potential as a novel antimicrobial agent.
Keywords: antimicrobial activity,
bacteriocin, DING protein, Halomonas sp. Strain M3
Abstrak
Sejenis halofil marin Halomonas sp. strain M3
telah dipencilkan daripada Selat Johor, Malaysia dan menghasilkan bakteriosin
CC yang bertindak sebagai ejen bakteriostatik. Pencirian bakteria ini
menunjukkan bahawa pertumbuhan yang optima pada suhu bilik, pH 8-8.5 dalam
nutrien media yang ditambah dengan 2.9% w/v NaCI untuk meniru keadaan air masin.
Kajian kestabilan menunjukkan bahawa bakteriosin CC adalah tahan haba (35°C-50 °C) dan stabil sepanjang 2 tahun
apabila disimpan dalam 0.02M Tris-HCI bersama 30-60% gliserol pada 4°C.
Kehilangan aktiviti telah dikesan selepas
rawatan enzim proteolitik, menunjukkan sifat protin dalam komposisi
antimikrob. Urutan asid amino bakteriosin CC telah diperolehi dengan
menggunakan degradasi Edman dan analisis MALDI-TOF, menunjukkan ciri protin
DING (D-I-N-G-G-G-A-T-L-P-Q-A-L-Y-Q) dalam saiz 38.9-kDa pada pI 6.8. Protin
ini membentuk satu set yang terpelihara dan edaran protin ini didapati secara
meluas dalam semua hidupan dengan aktiviti pengikatan-ligan dan enzim
hidrolitik, mencadangkan peranan yang mungkin ada dalam pengisyaratan sel dan
bio mineral dalam pemencilan DING yang lain. Menariknya, DING protin juga
terlibat sebagai agen anti-tumor pada manusia. Oleh itu, bakteriosin CC sebagai
ahli keluarga protin DING, perlu dikaji lagi untuk menyiasat sekiranya ia
berpotensi sebagai ejen antimikrob yang baru.
Kata kunci: aktiviti antimikrob,
bakteriosin, DING protin, Halomonas sp. Strain M3
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