79 research outputs found
The development of papain‐like protease from SARS‐CoV‐2, a potential drug target for antiviral screening: A review
No full textThe SARS‐CoV‐2 outbreak caused a global pandemic, claiming numerous lives and becoming this century’s most widespread life‐threatening disease. The virus relies on two specific enzymes to facilitate replication, 3‐chymotrypsin‐like protease (3CLPro) and papain‐like protease (PLpro). These enzymes are crucial in breaking down nonstructural polypeptides into functional proteins. PLpro with LXGG↓X recognition and cleavage sites also play a role in deubiquitylase (DUB) and delSGylase by cleaving after the double glycine residue of ubiquitin (Ub) and ISG15 as a mechanism to suppress the host’s innate immune response. Despite its important role in the viral infection cycle and the potential for drug discovery, no antivirals have been approved as PLpro inhibitors. Therefore, this review focuses on PLpro protein, its recombinant product development and purification, and its application as a protein target in drug discovery for COVID‐19 screening to develop effective COVID‐19 drugs
RNA THERAPEUTIC, PENDEKATAN BARU DALAM TERAPI GEN
No full textSome diseases, such as cancer, hereditary and genetic diseases, as well as viral infectious diseases, have been treated unsatisfied by the conventional therapy so far, and even more, by the gene therapy. Together with the pharmaceutical industries, researchers put their best effort to hunt some molecules that can be more favorable for such kind of therapy. After a pivotal study reported in May, 2001, it is certain that Ribonucleic acid (RNA) could effectively silence gene expression in mammalian cell line, so it was then proposed in 2004 the term RNA therapeutics. Antisense RNA therapy which came into the stage earlier seemed to be the one that can answer all the problems in knocking out the unwanted messenger in gene expression. RNA interference (RNAi) concept, which came later in around 2000, began to look like a possible contender. It was reported in some studies that RNAi seems to have some more advantages over both stronger gene-silencing effects and greater ease of use. However, the main obstacle of all kind of gene therapy is, undoubtedly, on the delivery of this molecule to enter the target cell, and mostly, to where it is needed most inside the body. Some studies on genetic material delivery system have been reported, and their progress has been discussed. Keywords: RNA therapeutics, antisense RNA, siRNA, terapi gen, gene silencing
Profiling Skin Microbiome in Healthy Young Adult Representing Javanese, Papuans, and Chinese Descent in Indonesia
No full textSkin serves as the first physical barrier and biological barrier by the colonization of commensal bacteria to prevent pathogen invasion. It was known that the disruption on normal commensal microbiota composition or dysbiosis causes skin diseases, while the skin microbiota diversity itself is influenced by several factors, one of them is ethnicity. This study shows the influence of ethnicity factor in Papuans, Javanese, and Chinese descent young adults living in Jakarta on skin microbiome profiles. The microbiota genomic DNA are extracted from the face skin samples and sequenced with Next Generation Sequencing method to be further analyzed. The result shows that individuals with the same ethnic background share similar skin microbiome characteristics. The greatest skin microbiome alpha diversity is shown by the Papuans and the Chinese descent the smallest. Ethnicity factor that shows statistically significant differences in interindividual dissimilarities are independent of other intriguing factors such as age, geographical location, etc. Therefore the ethnic origin of individuals especially from three ethnics above is a factor to be considered in skin microbiome research and the skin microbiota composition can be used for potential future applications
Characterization of Trypsin-Like Protease of Lactobacillus plantarum FNCC 0270
No full textTrypsin is an enzyme that has a unique mechanism of cutting peptide bonds specifically at the carboxyl side of lysine or arginine amino acids, with another amino acid. This study aims to analyze a trypsin-like protease (TLP) found in Lactobacillus plantarum FNCC 0270, by performing partial proteomic tests, i.e. MALDI-TOF/TOF, and standard bioinformatics tools. SDS-PAGE analysis showed 4 protein bands. Two bands of the (P1 and P2) showed molecular weights equivalent to 47.35 and 38.42 kD, each generating 8 and 11 peptide fragments respectively. According to information in www.ncbi.nlm.nih.gov/genbank/structures, the structure of serine protease HtrA (subs. plantarum L. plantarum ST-DT) consists of three domains. Using Clone Manager® software by aligning two sequences we obtained eleven. The Lactobacillus produces of the trypsin-like serine protease has 40-90% similarity. Using the Clustal W2 software we passed the 11 sequences through multiple alignments, and found that the isolate L. plantarum is closely related to L. buchneri, L. brevis, and L. malefermentans on the phylogenetic tree. Alignment analysis results showed that all 8 peptide fragments of band 1 and 11 peptide fragments of band 2, of the SDS-PAGE, were located in the active domain region of the fourth trypsin-like serine protease producing Lactobacilli
A NOVEL INTEGRON IN THE GENOME OF ESCHERICHIA COLI ISOLATED FROM INDONESIAN MONITOR LIZARD (VARANUS SPP).
No full textThe genotype of antibiotic resistance in natural isolates of Escherichia coli was determined through integron detection and characterization of the associated antibiotic resistance. E. coli SG2 isolated from Varanus salvator of Java demonstrated resistance to spectinomycin (50ng/ml) and streptomycin (SOng/ml). Integron detection indicated that eight isolates out of nine E. coli isolates possessed a conserved segment of the integron. Amplification of the inserted cassette of the integron in this SG2 isolate yielded a 1-kb DNA fragment. Sequence analyses indicated that this fragment was homologous with aad gene, which confirmed the resistance to spectinomycin/streptomycin. This is the first report on the presence of integron in the E. coli isolated from the environment. Key words: Integron / antibiotic resistance / Escherichia col
Bioactive fractions from Streptococcus macedonicus MBF 10-2 produced in an optimized plant-based peptone medium fermentation
No full textPostbiotic fractions of several lactic acid bacteria have potential as microbial therapeutics for skin health and may also appeal to consumers who wish to avoid animal-based products. We aim to establish the optimum plant-peptone fermentation of Streptococcus macedonicus MBF10-2, which possess Bacteriocin Like-Inhibitory Substance activity in our previous study, to produce bacterial bioactive fractions. We evaluate their potential antibacterial and antioxidant actions, and as well assess the preliminary safety for human skin application. Fermentation was carried out by using plant peptone modified MRS, i.e., soy peptone and Vegitone, a non-animal-carbon sources that substitute proteose peptone in MRS medium. Fractions of MBF10-2 lysate and cell-free supernatant were collected and processed as follows, i.e. cell disruption, fraction separation and fractions freeze-drying. Fractions were confirm for antibacterial properties by the agar well diffusion method and assess for antioxidant activity using DPPH, while safety assessment was carried-out by skin patch assay. Maximum growth of MBF10-2 achieved by fermentation in soy peptone- and in Vegitone-modified media was 9.00 and 7.99 g total cell mass, respectively. The antibacterial property of fractions was most effective against Micrococcus luteus T18. The lysate fraction exhibited a mild antioxidant potency (IC50 840 µg/mL), and all bioactive fractions were proven safe and non-allergenic for human skins. Strep. macedonicus MBF10-2 postbiotics bioactive fractions were indicated as being safe for topical application. This is the first report on the production of a safe Strep. macedonicus bioactive postbiotic possessing mild antibacterial and mild-to-weak antioxidant
Seleksi Galur-Galur Leuconostoc yang Mempunyai Aktivitas Bakteriostatin Terhadap Berbagai Bakteri Indikator
No full textLactid Acid Bacteria (LAB) are known to produce bacteriocins which have antimicrobial activity, and possessed to be developed as antibiotic complement. This study aimed to characterize bacteriocins activity from Leuconostoc strains isolated previously from local sources, and to optimize pH and incubation temperature as well. A well diffusion agar assay for zone inhibition method and bacteriocin potency assay performing minimum inhibition concentration (MIC) have been done. Bacterial indicators used in this study are Leu. mesenteroides TISTR 120, and JCM 6124, Staphylococcus aureus FNCC 0047, Listeria monocytogenes FNCC 0156, Escherichia coli FNCC 0183, Pseudomonas aeruginosa FNCC 0063, Salmonella typhi FNCC 0165 and Bacillus subtilis FNCC 0061. Catalase, Trypsin and Protease K were also used for confirmation test. Results revealed that both Leu. mesenteroides MBF2-5 and MBF7-17 possessed bacteriocin activity although against Leu. mesenteroides TISTR 120 and JCM 6124 indicators strains. The optimum pH for bacteriocin potency assay for both Leuconostoc strains MBF2-5 and MBF7-17 was pH 6, whereas the optimum incubation temperature was 32 oc with MIC value of 90% and 80%, respectively
HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS
No full textA transcriptional fusion for an Aeromonas caviae chitinase gene was constructed under the control of a constitutive promoter of the kanaraycin resistance gene (PKmR). The construct was inserted into a medium copy number broad host range plasmid vector to yield recombinant plasmid pAM340, which harbored transcriptional fusion PKmR- chi. Another transcriptional fusion, Ptac-chi, in a recombinant plasmid pAM630, was conducted as comparison. Triparental mating of E. coli carrying the recombinant plasmids with Pseudomotws fluorescens 5100, a phyllosphere bacterium, was performed. Pseudomonas fluorescens 5100 exconjugants were examined for constitutive expression of chitinase employing a spectrophotometric assay; they showed stronger chitin degradation activity than Escherichia coli transformants. Using a fungal antagonism plate assay, this chitinolytic P. fluorescens, however, could not inhibit selected phytopathogenic fungi. Keywords:   Aeromonas  caviae/ chitinase  gene/transcriptional  fusion/PKm\u27V  Vtac-chilPseudomonas fluorescen
TRANSGLYCOSYLATION ACTIVITY AND CHARACTERIZATION OF RECOMBINANT SUCROSE PHOSPHORYLASE FROM LEUCONOSTOC MESENTEROIDES MBFWRS-3(1) EXPRESSED IN ESCHERICHIA COLI
No full textObjective: Sucrose phosphorylase (SPase) is an enzyme that catalyzes the transfer of glucosyl to various acceptor molecules. Distinct types of SPaseshave been reported, and their transglycosylase activities have been shown to differ. In general, glycosylation is a process that is used to modifybioactive compounds. As such, glycosylation can increase the chemical stability of compounds and improve their characteristics such as reduce strongsmell and sour taste. We previously cloned recombinant SPase (SPaseWRS-3[1]) from Leuconostoc mesenteroides MBFWRS-3[1] in Escherichia coli.In the current study, we aimed to characterize SPaseWRS-3 and determine its transglycosylation activity using benzoic acid (BA), ascorbic acid, andkojic acid (KA).Methods: Expression analyses were conducted in lysogeny broth (LB) medium supplemented with tetracycline and expression was induced usingisopropyl-β-d-thiogalactopyranoside. The characteristics of the 56 kDa recombinant SPase (rec-SPase) were confirmed using sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). Rec-SPase activity was determined spectrophotometrically using sucrose as the substrate and NADPHas the end-product at 340 nm. Transglycosylation activity was evaluated using thin-layer chromatography (TLC) on silica gel plates.Results: Our results demonstrated that the rec-SPase had an activity of 98.52% relative to the reference SPase (ref-SPase). BA and KA were determinedto undergo glucosyl transfer by rec-SPase using ref-SPase, as observed with TLC. Our findings are consistent with those reported previously for theSPase isolated from L. mesenteroides.Conclusion: Recombinant SPase activity is comparable to reference SPase activity. Our study could be the initial study to deeply observe SPase activityin other substrates as well
Respon Galur Leuconostoc mesenteroides dan Weissella confusa terhadap pH Kondisi Pertumbuhan Menggunakan Antibiotik sebagai Indikator
No full textLactic acid bacteria is known to contribute to human health as exopolysaccharide producer, bacteriocin producer, live vaccins, starter in fermentation process and as probiotics. This study aimed to observe Leuconostoc mesenteroides and Weisella confusa strains response under manipulated growth condition for carbon source composition and on pH as a model, using various antibiotics as indicator. MRS and modified MRS (50% normal dextrose concentration) were used. Modification of pH representing acidic and alkaline condition were 4.6 and 9.0. Disc diffusion method was employed using six different antibiotics. Result showed that all strains grown in acidic standard MRS tend to be more sensitive to amoxicillin (Am), chloramphenicol (Ch) and polymyxin B (PoB), but more resistant to ciprofloxacin (Ci), while in the acidic modified MRS, the strains respond more sensitive to Am, Ch, Ci and PoB. When grown in alkaline standard MRS, all strains respond more sensitive to Am, Ci and PoB, but more resistant to Ch, while in alkaline modified MRS, they respond more resistant to Am and Ch but more sensitive to Ci and PoB. Most of the strains did not show inhibition against vancomycin and none of strains showed inhibition against sulphamethoxazole-trimethoprim under conditions applied.Bakteri asam laktat diketahui berkontribusi dalam kesehatan manusia sebagai penghasil eksopolisakarida, vaksin hidup, starter kultur dalam proses fermentasi dan sebagai probiotik. Penelitian ini bertujuan untuk mengamati respon galur Leuconostoc mesenteroides dan Weissella confusa di bawah kondisi komposisi sumber karbon dan pH yang termanipulasi dan dengan menggunakan indikator berbagai antibiotika. Media yang digunakan adalah MRS dan MRS termodifikasi (konsentrasi dekstrosa 50% dari normal). Modifikasi pH dilakukan pada kondisi asam (pH 4,6) dan basa (pH 9,0). Metode yang digunakan adalah difusi cakram menggunakan enam jenis antibiotik. Hasil menunjukkan bahwa semua galur dapat tumbuh di medium MRS + asam dan cenderung lebih sensitif terhadap amoksisilin (Am), kloramfenikol (Ch) dan polimiksin B (PoB), tetapi lebih resisten terhadap siprofloksasin (Ci). Dalam MRS termodifikasi + asam, galur cenderung menjadi lebih sensitif terhadap Am, Ch, Ci, dan PoB. Ketika ditumbuhkan pada MRS termodifikasi + basa, respon semua galur menjadi lebih sensitif terhadap Am, Ci, dan PoB, tetapi lebih resisten terhadap Ch, sedangkan dalam MRS termodifikasi + basa responnya menjadi lebih resisten terhadap Am dan Ch tetapi lebih sensitif terhadap Ci dan PoB. Kebanyakan galur tidak menunjukkan zona inhibisi terhadap vankomisin dan pada kondisi yang digunakan tersebut tidak satu galur pun menunjukkan zona inhibisi terhadap sulfametoksazol-trimetoprim
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