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Sains Malaysiana 45(3)(2016): 411–416 Collagen Binding Activity of Bacteria
Isolated from Pig and Cow Small Intestine (Aktiviti Pengikatan Kolagen Bakteria Dipencilkan
daripada Usus Kecil Babi dan Lembu) NUR FADHILAH
KHAIRIL
MOKHTAR1,2*,
RAHA
ABDUL
RAHIM2,
AMALIA
MOHD
HASHIM2
& SHUHAIMI MUSTAFA1,2 1Halal Products
Research Institute, Putra Infoport, Universiti Putra Malaysia, 43400
Serdang, Selangor Darul Ehsan, Malaysia 2Faculty of Biotechnology
and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang,
Selangor Darul Ehsan, Malaysia Received: 31 March
2015/Accepted: 28 August 2015 ABSTRACT Bacterial adherence to connective
tissue, especially to collagen has been vastly known for their invasive
and infectious activities. However, the ability to exploit the unique
and specific interactions between bacteria and collagen as a novel
approach in detection of placental collagen has never been explored.
This study aimed to determine bacteria with binding specificity
to placental collagen (Type IV) derived from human and sheep. In
order to do this, total bacteria from small intestines of pig and
cow were isolated and their ability to bind to Type IV placental
collagen (human and sheep) was determined. Interestingly, three
bacterial samples; P5, P9 (pig small intestine origin) and B7 (cow
small intestine origin) were found to be able to bind strongly to
the placental collagen. The bacterial binding to human placental
collagen was however, diminished after the bacteria were treated
with trypsin, proteinase K (for removal of surface protein) and
guanidine hydrochloride (for S-layer removal), suggesting that the
interaction of these bacteria to placental collagen was promoted
by protein(s) present at the bacterial surface. In addition, significant
reduction of placental collagen-binding ability of the bacteria
pre-incubated with soluble human placental collagen showed that
there is a specific interaction between the bacteria and collagen.
P5, P9 and B7 bacteria were found to share 95-97% 16S rRNA sequence
similarity to Enterococcus
faecalis ZL, Enterococcus hirae ss33b and Enterococcus
faecium M3-1, respectively. The results presented here may facilitate
future studies in identifying bacterial surface protein(s) responsible
for the specific binding of bacteria to collagen and opens new opportunity
to utilize the protein(s) for the detection of placental collagen
in nutraceutical and food supplements. Keywords: Bacterial binding;
placental collagen; surface protein ABSTRAK Pelekatan bakteria kepada tisu
penghubung, terutamanya kolagen telah diketahui dengan meluas untuk
aktiviti invasif mereka. Walau bagaimanapun, dalam kajian ini, keupayaan
untuk mengeksploitasi interaksi yang unik dan khusus antara bakteria
dan kolagen sebagai pendekatan baru dalam pengesanan kolagen plasenta
telah dikaji. Bakteria daripada usus kecil khinzir dan lembu telah
diasingkan dan keupayaan mereka untuk mengikat kepada kolagen plasenta
‘Type’ IV (manusia dan biri-biri) telah dikenal pasti. Menariknya,
tiga sampel bakteria; P5, P9 (daripada usus kecil babi) dan B7 (daripada
usus kecil lembu) didapati mampu mengikat teguh kepada kolagen plasenta.
Bagaimanapun, keupayaan bakteria untuk mengikat plasenta kolagen
manusia berkurangan selepas bakteria dirawat dengan tripsin, proteinase
K (untuk pembuangan protein permukaan) dan guanidin hidroklorida
(untuk pembuangan lapisan S). Ini menunjukkan bahawa interaksi bakteria
untuk kolagen plasenta dipromosikan oleh protein yang wujud pada
permukaan bakteria. Selain itu, pengurangan yang ketara daripada
keupayaan bakteria untuk mengikat kolagen plasenta selepas dieram
dengan kolagen larut plasenta menunjukkan bahawa terdapat interaksi
tertentu antara bakteria dan kolagen. Bakteria P5, P9 dan B7 didapati
mempunyai 95-97% persamaan daripada segi jujukan 16S rRNA masing-masing
dengan Enterococcus faecalis ZL, Enterococcus hirae
ss33b dan Enterococcus faecium M3-1. Hasil daripada kajian
ini boleh digunakan sebagai asas kajian lanjut untuk mengenal pasti
protein permukaan bakteria tertentu yang bertanggungjawab mengikat
kolagen dan membuka peluang baru untuk menggunakan protein tersebut
bagi pengesanan kolagen plasenta dalam makanan tambahan dan nutraseutikal. Kata kunci: Kolagen plasenta; pengikatan bakteria; protein permukaan REFERENCES
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