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Journal : Microbiology Indonesia

The Role of the First 14 Amino Acids of Mature M1 Protein of Streptococcus pyogenes on Fibronectin-Binding Activity and Dimer Formation ROGA FLORIDA KEMBAREN; ADAM REZA GANJARA; VALENTINA YURINA; DEBBIE SOEFIE RETNONINGRUM
Microbiology Indonesia Vol. 4 No. 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1672.376 KB) | DOI: 10.5454/mi.4.1.5

Abstract

Streptococcus pyogenes is one of the most important human pathogens which express a multi-facet of virulence factors on its cell surface. One of the virulence factors that has been intensively-studied is the M protein that binds several human proteins. M1 protein, a member of the M protein family, was previously found to bind human fibronectin (Fn), an activity that is responsible for bacterial internalization. A structural study showed that this protein consists of four regions: A, B, S, and C. The study was intended to investigate the role of the first 14 amino acid residues located at the non-helical region of M1 protein in binding Fn, and its ability to form a dimer. The DNA fragment encoding for the ABS protein lacking its first 14 amino acids (ABSD14aa) was cloned into pET-16b, overexpressed in Escherichia coli BL21(DE3), and the protein was purified by affinity chromatography. The purified protein was characterized by sodium dodecyl sulphate polyacrylamide gel electrophoresis and the Fn-binding activtiy was assayed by enzyme linked immunosorbent assay. The result indicated that the M1 lacking its first 14 amino acids retains its dimerization and Fn-binding activities.
Molecular Analysis of Immune-Escape Mutants of Hepatitis B Virus from Local Clinical Samples CHANDRA JINATA; ERNAWATI ARIFIN GIRI-RACHMAN; DEBBIE SOEFIE RETNONINGRUM
Microbiology Indonesia Vol. 6 No. 1 (2012): March 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (379.113 KB) | DOI: 10.5454/mi.6.1.2

Abstract

Small hepatitis B surface antigen (sHBsAg) is used as a component of hepatitis B vaccine. Even though this vaccine is known to be effective in preventing hepatitis B disease, natural mutation may induce Hepatitis B Virus (HBV) to form immune-escape mutant. This mutant is not only capable of infecting hepatitis B-vaccinated people, but also causing commercial diagnostic assay failure. Immune-escape mutant is generally detected from amino acid change at Major Hydrophilic Region (MHR) of sHBsAg while the change occurred outside the region may also lead to immune-escape mutant formation. This research was aimed to investigate the presence of HBV immune-escape mutants in local clinical samples in Indonesia. sHBsAg gene of seventeen HBV samples from local patients were amplified by polymerase chain reactions then subjected to two-directional sequencing. The DNA sequences later were analyzed by bioinformatics programs. Fifteen out of seventeen samples were genotype B and subtype adw2, while the other two were genotype C and subtype adrq+. Among fifteen genotype B samples, twelve of them were not immune-escape mutants, two were immune-escape mutants that have been previously reported (Gln129Arg and Met133Leu), and one was a mutant outside MHR that has not been previously reported as an immune-escape mutant (Tyr161Ser). Both samples of genotype C group were not immune-escape mutants. As conclusion, by investigating seventeen local clinical HBV samples, it was known that two of seventeen samples were confirmed as immune-escape mutants and one of seventeen samples was a mutant outside MHR.
16S rDNA-Based Identification of Novel Superoxide Dismutase Producing Bacteria Isolated from Indonesia ANA INDRAYATI; VALENTINA YURINA; LARAS AJENG PITAYU; DEBBIE SOEFIE RETNONINGRUM
Microbiology Indonesia Vol. 5 No. 2 (2011): June 2011
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (145.387 KB) | DOI: 10.5454/mi.5.2.6

Abstract

Superoxide dismutase (SOD) has therapeutic importance because of its antioxidant activity and protects cells from reactive oxygen species attack. This research was intended to screen bacteria isolated from Indonesia for producing novel SODs and to identify the producers using 16S rDNA approach. Intracellular proteins were each extracted and assayed for their inhibition reduction activity by colorimetric method and by zymography for the presence of SOD protein band(s). For species identification, each of 16S rDNAgenes was amplified by polymerase chain reaction from genomicDNAfollowed by sequencing, BLAST, multiple alignment and phylogenetic analyses. All 16 intracellular proteins gave inhibition reduction percentage in the range of 15 to 70% and in zymography, their SOD profiles were quite diversed with at least one intenseSOD band present in most isolates. The SOD producers were assigned to three species, Flavobacterium okeanokoites, Escherichia fergunosii, and E. coli, and to four genera, Pantoea, Escherichia,  Bacillus, and Pectobacterium. The remaining five were grouped in gamma-proteobacterium cluster and two formed a cluster with Pseudomonas. Three marine and four soil isolates could be attractive candidates for novel SODs based on unique properties of SOD producers. In conclusion, 16s rDNA-based identification of bacteria isolated from Indonesia reveals that seven isolates might be attractive candidates for novel SOD producers to be applied in pharmaceutical fields in the future.