Vol 20, No 3 (2016) > Chemical Engineering >

Expression and the Functional Study of Fusion Proteins α-Amylase and Hemolysin- αas an Application in Biofilm Polysaccharide Degradation

Gede Yuda Sugiarta 1 , Anggoro Wiseso 2 , Siska Yuliana Sari 3 , Etri Dian Kamila 2 , Vanessa Geraldine 2 , Diana Christina 2 , Muhammad Hanifi 1 , Dwiantari Satyapertiwi 2 , Robby Hertanto 1 , Budiman Bela 4 , Masafumi Yohda 5 , Muhamad Sahlan 2

Affiliations:

  1. Faculty of Medicine, Universitas Indonesia, Depok 16424, Indonesia
  2. Research Centre for Biomedical Engineering, Department of Chemical Engineering, Faculty of Engineering, Universitas Indonesia, Depok 16424, Indonesia
  3. Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Depok 16424, Indonesia
  4. Department of Clinical Microbiology, Faculty of Medicine, Universitas Indonesia, Jakarta 10320, Indonesia
  5. Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan

 

Abstract: Biofilm is an aggregate of consortium bacteria that adhere to each other on a surface. It is usually protected by the exopolysaccharide layer. Various invasive medical procedures, such as catheterization, endotracheal tube installation, and contact lens utilization, are vulnerable to biofilm infection. The National Institute of Health (NIH) estimates 65% of all microbial infections are caused by biofilm. Periplasmic α-amylase (MalS) is an enzyme that hydrolyzes α-1, 4-glicosidic bond in glycogen, starch, and others related polysaccharides in periplasmic space. Another protein called hemolysin-α (HlyA) is a secretion signal protein on C terminal of particular peptide in gram negative bacteria. We proposed a novel recombinant plasmid expressing α-amylase and hemolysin-αfusion in pSB1C3 which is cloned into E.colito enable α-amylase excretion to extracellular for degrading biofilm polysaccharides content, as in starch agar. Microtiter assay was performed to analyze the reduction percentage of biofilm by adding recombinant E.coli into media. This system is more effective in degrading biofilm from gram positive bacteria i.e.: Bacillus substilis(30.21%) and Staphylococcus aureus (24.20%), and less effective degrading biofilm of gram negative i.e.: Vibrio cholera (5.30%), Pseudomonas aeruginosa (8.50%), Klebsiella pneumonia (6.75%) andE. coli (-0.6%). Gram positive bacteria
have a thick layer of peptidoglycan, causing the enzyme to work more effectively in degrading polysaccharides.
Keywords: Alpha amylase, Biofilm, Hemolysin-α, Recombinant plasmid
Published at: Vol 20, No 3 (2016) pages: 127-131
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