Sains Malaysiana 45(7)(2016): 1057–1062
Purification
of Glutathione Transferases (GSTs) from Identified Rhizospheric Bacteria
(Penulenan
Glutation Transferases (GSTs) daripada
Bakteria Rizosfera Dikenal Pasti)
KHANOM SIMARANI*, WAN HAIZATUL AKMAL CHE YUSOFF
& ZAZALI ALIAS
Institute of Biological Sciences, Faculty of Science,
University of Malaya, 56100 Kuala Lumpur
Malaysia
Diserahkan: 12 Mac 2015/Diterima: 27 Januari 2016
ABSTRACT
The glutathione S-Transferase (GST)
enzyme plays an important role in cellular detoxification. This multifunctional
enzyme is involved in Phase II detoxification pathways that protect cellular
macromolecules from being attacked by harmful compound. The study is an attempt
to isolate glutathione transferase-expressing bacteria from the rhizospheric
soil of selected herbal plants. Screening showed nine positive isolates out of
twelve bacterial samples from a large microbial population in our soil
collection. Crude extract from strain E1 which was isolated from Piper sarmentosum (Kadok) showed the highest
specific activity against 1-chloro-2, 4-dinitrobenzene substrates (5.78 × 10-06 μmol/min/mg). Based on the carbon utilization of
E1 assessed using Biolog system, the strain was identified as Comamonas
testosterone E1. Glutathione S-transferase purification using GST trap
yielded two distinct subunits with molecular weights of 23 and 24 kDa as
visualized on 1D SDS-polyacrylamide gel
electrophoresis. The purified GST showed reactivity towards
1-chloro-2, 4-dinitrobenzene, 1, 2-dichloro-4-nitrobenzene and ethacrynic acid
with specific activity of 0.264 ± 0.038 nmol/min/mg and 0.056 ± 0.002
nmol/min/mg and 10.500 ± 3.130 nmol/min/mg, respectively. However, no activity
was detected against p-Nitrobenzyl chloride, Sulfobromophthalein,
trans-4-phenyl-3-butene-2-one, hexa-2, 4- dienal, trans-hepta-2, 4-dienal and
trans-oct-2-enal in the study.
Keywords: Comamonas testosterone; GST-expressing
bacteria; Piper sarmentosum;
rhizospheric soil; xenobiotic
ABSTRAK
Enzim glutation S-Transferase (GST)
memainkan peranan yang penting dalam detoksifikasi bersel. Enzim pelbagai fungsi ini
terlibat dalam Fasa II laluan detoksifikasi yang melindungi sel makromolekul
daripada diserang oleh sebatian berbahaya. Kajian ini
merupakan cubaan untuk memencilkan bakteria glutation transferase-nyata dari
tanah rizosfera beberapa tanaman herba. Daripada
keseluruhan dua belas sampel bakteria dari populasi mikroorganisma yang
terdapat dalam sampel tanah terkumpul yang diuji, sembilan sampel didapati
berpotensi. Ekstrak mentah daripada strain E1 yang telah diasingkan
daripada pokok herba Piper sarmentosum (Kadok)
menunjukkan jumlah aktiviti spesifik yang tinggi terhadap substrat 1-chloro-2,
4-dinitrobenzena (5.78 × 10-06 μmol/min/mg). E1
dikenal pasti sebagai Comamonas testosterone berdasarkan penggunaan
karbon yang dinilai menggunakan sistem Biolog. Penulenan glutation
S-transferase (GST) menggunakan perangkap GST menghasilkan
dua subunit berbeza dengan berat molekul masing-masing 23 dan 24 kDa seperti
ditunjukkan dalam 1D SDS-poliakrilamida gel elektroforesis. GST tulen menunjukkan reaktiviti terhadap 1-kloro-2, 4-dinitrobenzena
(CDNB),
1, 2-dikloro-4-nitrobenzena (DCNB) dan asid etakrinik (EA)
iaitu dengan jumlah aktiviti spesifik masing-masing sebanyak 0.264 ± 0.0382,
0.056 ± 0.002 dan 10.500 ± 3.130 nmol/min/mg. Walau bagaimanapun, aktiviti
terhadap p-Nitrofenil klorida (NBC), sulfobromoftalein (BSP),
trans-4-fenil-3-butena-2-satu (PBO), hexa-2, 4- dienal,
trans-hepta-2, 4-dienal dan trans-oct-2-enal tidak dikesan dalam kajian ini.
Kata kunci: Comamonas testosterone;
bakteria GST-nyata; Piper sarmentosum;
tanah rizosfera; xenobiosis
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*Pengarang
untuk surat-menyurat, email: hanom_ss@um.edu.my
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