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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 Diserahkan: 31
Mac 2015/Diterima: 28 Ogos 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
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*Pengarang untuk surat-menyurat; email:
nur021057@gmail.com
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