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All Journal HAYATI Journal of Biosciences Microbiology Indonesia Jurnal Matematika & Sains Pharmauho: Jurnal Farmasi, Sains, dan Kesehatan Jurnal Farmasi Indonesia YARSI Medical Journal

DEBBIE SOEFIE RETNONINGRUM

School of Pharmacy, Institut Teknologi Bandung

Author-ID : 359888

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Earth & Planetary Sciences Immunology & microbiology Mathematics Medicine & Pharmacology

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Articles

Optimasi Proses Overproduksi, Pemurnian dan Karakterisasi Protein Mga Sebagai Molekul Target Untuk Pencegahan Infeksi oleh Streptococcus Pyogenes Muhaimin Muhaimin; Oei Ban Liang; Enny Ratnaningsih; Endang Purwantini; Debbie Soefie Retnoningrum
Jurnal Matematika & Sains Vol 8, No 3 (2003)
Publisher : Institut Teknologi Bandung

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Optimization of overproduction, purification and characterization of Streptococcus pyogenesâs Mga recombinant protein from Escherichia coli pMALMga carrying mga49 gene had been carried out. The overproduction optimization was conducted by culturing the Escherichia coli pMALMga of OD600 = 0,7 in LB medium with or without glucose, and induced by 0,3 mM IPTG for a certain period of time. The protein produced was MBP-Mga fusion protein with a molecular weight of about 104 kDa. The purification was conducted by affinity chromatography using amylose ligands at pH 7,4 and continued by SDS-PAGE. One band identified at 104 kDa showed that the MBP-Mga fusion protein was pure enough for further analysis. The MBP-Mga protein was then injected into rabbit to produce polyclonal antibodies. The antibody development was monitored by dot blot and Western blot. The antibodies produced gave a positive reaction to MBP-Mga fusion protein. Adsorption and precipitation of the polyclonal antibodies which gave a positive reaction to MBP protein was done by adding pure MBP protein (56 kDa) produced by Escherichia coli pMALc2, and purified by affinity chromatography using amylose ligands at pH 7,4. The results showed that the antibodies gave a positive reaction to Mga recombinant protein and a negative reaction to MBP recombinant protein.

Extracellular Products of Streptococcus pyogenes and Their Involvement in Pathogenesis Debbie Soefie Retnoningrum
Jurnal Matematika & Sains Vol 14, No 2 (2009)
Publisher : Institut Teknologi Bandung

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Streptococcus pyogenes or group A streptococcus (GAS) is an exclusive human pathogen. To be a successful pathogen, this pathogen is equipped with various surface-exposed and secreted virulence factors. The functions of secreted virulence factors are particularly important since they interact with host components to establish infections and cause diseases in human. They include a number of proteases, DNase, superantigens, and plasminogen activator. How these secreted factors interact with host protein(s) define this pathogens ability to bring out various diseases. Several proteases act independently to target immunoglobulin molecules in order to evade host defense system and modulate host proteins to induce host-mediated damages. Besides producing proteases, many pathogenic strains of GAS also produce DNase; however, its involvement in host pathogenesis remains elusive. Superantigen, another secreted protein is responsive for serious host-destruction by bypassing antigen presentation to induce massive production of cytokines. GAS also secretes a plaminogen activator, streptokinase that is crucial for invasiveness. All together, secreted products of this pathogen work in concert to pinpoint different targets in order to destroy and or to disable human defense system and cause host damages.

Penentuan Aktivitas Siklisasi-Î² Siklodekstrin Glukosiltransferase Wild Type, H43K, P84Y, Y93F, Y188L, dan S468F dari Bacillus sp. A2-5a terhadap Pati Terlarut Rostinawati, Tina; Sumirtapura, Yeyet Cahyati; Elfahmi, Elfahmi; Retnoningrum, Debbie Soefie
Pharmauho: Jurnal Farmasi, Sains, dan Kesehatan Vol 2, No 1 (2016): Pharmauho
Publisher : Fakultas Farmasi Universitas Halu Oleo

Siklodekstrin (CD) merupakan oligosakarida yang terdiri atas enam sampai delapan glukosa yang dihubungkan dengan ikatan 1,4 glikosida (CD-Î±, -Î² dan -Î³). Molekul ini memiliki kemampuan membentuk kompleks inklusi dengan suatu molekul â€tamuâ€ (guest) yang bersifat hidrofobik. CD membentuk cincin yang rongganya bersifat hidrofob namun permukaan luarnya bersifat hidrofil sehingga memungkinkan molekul tamu yang tidak larut air untuk berkompleks di dalam rongganya. Senyawa ini diaplikasikan secara luas pada berbagai industri farmasi, bioteknologi, makanan, kimia, tekstil,Â pertanian dan juga untuk perlindungan lingkungan. CD dihasilkan melalui reaksi enzimatis siklodekstrin glikosiltransferase (CGTase) terhadap pati sebagai substrat. CGTase yang berasal dari Bacillus sp. A2-5a (BA2-5a) merupakan CGTase-Î² yang menghasilkan CD-Î² sebagai produk utamanya dengan sebagian kecil CD-Î± dan selebihnya CD-Î³. Pada penelitian sebelumnya telah dihasilkan CGTase BA2-5a mutan melalui substitusi asam amino pada posisi 43, 84, 93, 188 dan 468 yang menghasilkan CGTase BA2-5a H43K, P84Y, Y93F, Y188L dan S468F. Pada penelitian ini ditentukan aktivitas siklisasi-Î² CGTase BA2-5a wild type dan CGTase mutan H43K, P84Y, Y93F, Y188L dan S468F. Tujuan penelitian ini untuk mengetahui aktivitas siklisasi-Î² CGTase wild type dan pengaruh substitusi asam amino pada posisi 43, 84, 93, 188 dan 468 terhadap aktivitas tersebut. Hasil penelitian menunjukkan aktivitas siklisasi-Î² CGTase wild type, CGTase H43K, CGTase P84Y, CGTase Y93F, CGTase Y188L dan CGTase S468F adalah 5,01; 2,14; 2,78; 1,77; 0,62; dan 3,37 unit/ml. Pada penelitian ini juga ditentukan aktivitas spesifik CGTase wild type, CGTase H43K, CGTase P84Y, CGTase Y93F, CGTase Y188L dan CGTase S468F sebagai berikut : 65,6; 41,2; 53,8 29,1; 11,0; dan 57,6 (unit/mg). Substitusi asam amino pada CGTase BA2-5a menunjukkan penurunan aktivitas siklisasi-Î².Kata kunci: BA2-5a, CGTase wild type, substitusi, CGTase mutan, aktivitas siklisasi-Î²

Karakterisasi Molekular Fragmen Gen mexB Isolat Pseudomonas aeruginosa Multiresisten Badaruddin, Fatmawaty; Supardi, Imam; Chatib Warsa, Usman; Soefie Retnoningrum, Debbie
Jurnal Kedokteran YARSI Vol 16, No 1 (2008): JANUARI - APRIL 2008
Publisher : Lembaga Penelitian Universitas YARSI

Antibiotics have been widely used in the treatment of infectious diseases. However, their effectiveness has been questioned due to the tendency of some bacterial resistance to antibiotics. Pseudomonas aeruginosa among others has been known to be resistant to several antibiotics due to its MexABOprM efflux pump. Perhaps, the nucleotide sequence of its mexB gene fragment has changed followed by changes in amino acid sequence leading to alteration of the substrate recognition site. This alteration causes disability of antibiotics to recognize it and they are pumped out from the bacterial cell causing decrease in its inhibition concentration. An observasional study was performed using four P. aeruginosa isolates (A,B,C and D) taken from four laboratories in Bandung, and the sensitivity test for several antibiotics (tetracyclin, ampicyllin, amoxicyllin-clavulanat, kanamycin, ciprofloxazin, trimetoprim-sulphametoxazol, chloramphenicol dan eritromycin), was performed using Kirby-Bauer method. The Minimum Inhibitory Concentrations (MICs) for 4 isolates were 20.57-39.07 mg/ml for erithromycin, 29.35-48.57 mg/ml for kanamycin, 30.35-68.75 mg/ml for tetracyclin, 45.57-97.50 mg/ml for ampicyllin, 23.69-97.50 mg/ml for chloramphenicol, 25.82-59.56 mg/ml for amoxcyllinclavulanat,21.88-79.00 mg/ml for trimetoprim sulphametoxazol, and 20.58-56.97 mg/ml for ciprofloxazin. The increasing of MIC to each antibiotic was shown for 4 isolates of P. aeruginosa multiresistant to several antibiotics being studied. PCR technique was used to detect mexB gene fragment asumed as the substrate recognition site. The percentage of homology between the nucleotide sequence of mexB multiresistant P. aeruginosa and mexB P. aeruginosa producing siderophore pioverdin (Acc. No. L11616, NCBI) showed 96%, 100%, 97%, and 96% homology for P. aeruginosa A,B,C and D respectively. Employing DNAstar program, fragment variant of mexB gene of 4 multiresistant isolates A, B, C and D was detected. This variation lead to amino acid substitution of Gly-417->Ser, Glu-417->Gln, Thr-424->Pro, Tyr-328->Phe, Asp-328->His for P. aeruginosa A,B,C and D respectively, along with the change of their secondary structure, that changed the functional protein of MexB.

Integration Stability of sHBsAg-Multi Expression Cassettes in Pichia pastoris GS115 during Methanol Induction Patricia Gita Naully; Neni Nurainy; Elvi Restiawaty; Dessy Natalia; Debbie Soefie Retnoningrum; Wardono Niloperbowo; Ernawati Arifin Giri-Rachman
HAYATI Journal of Biosciences Vol. 27 No. 4 (2020): October 2020
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.27.4.283

Hepatitis B is the major health problem worldwide including in Indonesia. Vaccination is the best prevention strategy for the disease. For the purpose of vaccine development and to decrease drug import, production of Hepatitis B Virus (HBV) small surface antigen (sHBsAg) from Indonesian HBV subtype is needed. The recombinant protein production can be conducted by integrating multi expression cassettes of sHBsAg gene in Pichia pastoris chromosome using gene replacement method. Such integration method turns out to allow loss of foreign gene from chromosome by excisional recombination-mediated looping out. This research was aimed to determine integration stability of four copies of sHBsAg expression cassette in P. pastoris GS115 chromosome inducted with 2% methanol in FM22 medium. The methanol induction was conducted twice at 63-h and 75-h. Integration stability determination was conducted qualitatively using PCR and quantitatively using qPCR absolute quantification. A band of 208 bp with similar intensity was observed after amplification of genomic DNA. All samples generated the same Ct value of around 22 with four copies of sHBsAg gene per genome. The result from this experiment shows that integration of four copies of sHBsAg expression cassette in P. pastoris GS115 chromosome is stable during methanol induction.

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

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

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

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.

Pengaruh Superoksida Dismutase Rekombinan Staphylococcus equorum Terhadap Viabilitas Sel dan Deposisi Kolagen Pada Sel Fibroblas 3T3 Yang Dipaparkan UVA Ana Indrayati; Sukmadaja Asyarie; Tri Suciati; Debbie Soefie Retnoningrum
Jurnal Farmasi Indonesia Vol 13 No 1 (2016): Jurnal Farmasi Indonesia
Publisher : Fakultas Farmasi Universitas Setia Budi

UVA is the main external factors causing skin aging. UVA increase matrix metalloproteinase (MMP) synthesis that degrade collagen indirectly through reactive oxygen species. Superoksida dismutase (SOD) catalyzes the conversion of superoxide anions to hydrogen peroxide and oxygen. SOD can be used as a cosmeticâ€™s component to prevent skin aging. The objective of this research to determine rSOD S. equorum ability to increase collagen deposition using fibroblast 3T3 against UVA. The research was initiated by the confirmation of E. coli BL21(DE3) carrying pJExpressÂ®414-sod. rSOD was overproduced in E. coli BL21(DE3) by IPTG induction to a final concentration of 1 mM for 4 hours at 37oC. rSOD purification was done using an Ni-NTA affinity column with gradient imidazole concentrations for elution. Various concentrations of rSOD was added to fibroblast 3T3 cell culture after UVA irradiation to determine its role in collagen deposition. The effect of rSOD was analyzed by fibroblast viability using Alamar blue and collagen deposition with picro sirius red. The results showed that fibroblast cell viability exposed with UVA for 45 minutes in the presence of 4, 8, and 16 unit rSOD was not significantly affected, however, the collagen deposition was significantly increased about 3.02%; 20.03% and 35.75% respectively compared to cell without rSOD. This result indicates that rSOD is a good candidate for an anti photoaging agent.

Co-Authors ADAM REZA GANJARA Ana Indrayati CHANDRA JINATA Chatib Warsa, Usman Dessy Natalia Elfahmi Elfahmi, Elfahmi Elvi Restiawaty Endang Purwantini Enny Ratnaningsih ERNAWATI ARIFIN GIRI-RACHMAN Ernawati Arifin Giri-Rachman Fatmawaty Badaruddin Imam Supardi LARAS AJENG PITAYU Muhaimin Muhaimin Neni Nurainy Oei Ban Liang Patricia Gita Naully ROGA FLORIDA KEMBAREN Sukmadaja Asyarie Sumirtapura, Yeyet Cahyati Tina Rostinawati Tri Suciati VALENTINA YURINA VALENTINA YURINA Wardono Niloperbowo

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