Cellulolitic Activities Of Actinomycetes Isolated From Soil Rhizospere of Waigeo, Raja Ampat, West Papua

Seven actinomycetes isolated from Waigeo’s soil rhizospere have been characterized in growth, cellulase activities and pH. These actinomycetes were chosen based on their capabilities in cellulose degradation which were recognized by previous study. Those isolates are belonging to Streptomyces and Actinoplanes genera based on morphological and molecular analyses. The growths of actinomycetes were investigated based on spectrophotometric approach and cellulase activities were conducted based on reduction sugar. Cellulase activities, cell growth, and pH had significant correlation based on statistical analysis.  The highest enzyme activities was acquired from  Streptomyces bobili LIPIMC-A-283 (0,519 µmol mL-1 minute-1) at 96 h of incubation. This study provided important basic information on potential cellulolitic alternative from actinomycetes group, as consideration on choosing low-cost technique using microbial agent in cellulose treatment

Studi Kelimpahan Aktinomisetes Tanah Dan Hubungannya Terhadap Enzim Selulase, Amilase, Total Karbon Dan Nitrogen Hutan Pasca Kebakaran Bukit Bangkirai Kalimantan Timur

Soil Actinomycetes Population, Enzymes Activity, and its Relation with Carbon and NitrogenContent, in Bukit Bangkirai, East Kalimantan. Bukit Bangkirai is one of the tropical forest inIndonesia have been exposed with intense forest fire. The affected forest is subjectivelydivided into three level of damages, heavily damage forest (HD), low damage forest (LD) andcontrol (K). The objective of this research was to observe the abundance of Actinomyceteswhich have important role in ecological process. Through decompotition of organic materialsand nutriens cycle. Actinomycetes were isolated and enumerated by SDS-YE method. CFU/ gsoil (x 104) in K, HD and LD are 41,86 ± 25,52, 16,09 ± 5,70 and 18,96 ± 4,19 respectively. Amylaseand cellulase were determined by DNS method. Carbon and Nitrogen total were determinatedby CN analyzer. The different of amylase, cellulase activities and abundace of Actinomycetesbetween HD, LD, and Control plot were not significant. However, carbon and nitrogen totalare different. LD plot has the highest carbon and nitrogen total, followed by control and HDplot. There has no significant different among plot observed may indicate microbial communitiesof soil in Bukit Bangkirai have been recovered

抗赤痢アメーバ新規薬剤開発を目的とした補酵素A生合成経路の解明

筑波大学 (University of Tsukuba)201

Identifikasi Molekular Dan Karakterisasi Morfo-Fisiologi Actinomycetes Penghasil Senyawa Antimikroba

The objectives of study were to identify antimicrobial producing Actinomycetes using 16S rDNA analyses and morphology and physiology characteristics. Eight Actinomycetes strain with the higest antibacterial and antifungal activity were selected and identified using six primers (20F, 520F, 920F, 1500R, 920R, and 520R). Morphological observation and physiology analyses were performed to the selected strain to accurately identify the strains. Morphological characters observed were aerial mycelium, spore chain, colony form, and pigment production. Physiological characterizations were antimicrobial properties, growth temperature, pH tolerance, salinity concentration for growth, sugars assimilation, and some enzymes production (arginine dihydrolase, urease, ß-glucosidase, protease, ß-galactosidase). Based on homology search by BLAST program and phylogenetic tree analyses, all of isolates were identified as the genus Streptomyces. They belong to eight different spesies. Isolates RC-SS-37-4, RC-SS-37-16 and BL-22-3 have been identified as Streptomyces costaricanus (100 %), Streptomyces costaricanus (99.8 %) and Streptomyces parvulus (98.6 %), respectively. Five isolates were identified as Streptomyces spp. (BL-36-1, BL-20-2, BL-14-2, BL-22-1 and BL-06-5) and can be presumed as new species because of the low homology value to their closest related spesies

Phosphate solubilizing activities of Actinomycetes isolated from Waigeo, Raja Ampat islands, West Papua

Actinomycetes is a major microbial group observed in soil, and contributes to nutrient cycling. This study is intended to verify physiological characters and phosphate solubilizing ability of Actinomycetes isolated from soil of Raja Ampat, West Papua. Most of isolates (RCW16-9, RCW16-8, W5-6, RCW25-1, RCW26-5, W28-4, W3-1, W3-7, W17-7, and W10-1) belonged to Streptomyces genera. The isolates produce clear zone in Pivoskaya after 3 days incubation. The liquid growth of this isolate rapidly utilizes glucose, and after 24 days of incubation almost 95% glucose was consumed. Decrease of pH from 6.1 to 4.3 may stimulate dissolution of calcium phosphate, and about 21 mg/L-P was observed in bulk solution. An increase of phosphomonoesterase activity during incubation is concomitant with the release of orthophosphate into bulk solution. Acidity of cultures increased may stimulate solubilization of calcium phosphate. Most strains produce phosphomonoesterase enzyme, indicating that actinomycetes are important soil microbes responsible for mediation and stimulation of both inorganic and organic phosphate dissolution. Physiological and biomass growth character of phosphate solubilizing actinomycetes could be important taxonomic indicator for identifying and grouping soil actinomycetes

Biochemical, Metabolomic, and Genetic Analyses of Dephospho Coenzyme A Kinase Involved in Coenzyme A Biosynthesis in the Human Enteric Parasite Entamoeba histolytica

Coenzyme A (CoA) is an essential cofactor for numerous cellular reactions in all living organisms. In the protozoan parasite Entamoeba histolytica, CoA is synthesized in a pathway consisting of four enzymes with dephospho-CoA kinase (DPCK) catalyzing the last step. However, the metabolic and physiological roles of E. histolytica DPCK remain elusive. In this study, we took biochemical, reverse genetic, and metabolomic approaches to elucidate role of DPCK in E. histolytica. The E. histolytica genome encodes two DPCK isotypes (EhDPCK1 and EhDPCK2). Epigenetic gene silencing of Ehdpck1 and Ehdpck2 caused significant reduction of DPCK activity, intracellular CoA concentrations, and also led to growth retardation in vitro, suggesting importance of DPCK for CoA synthesis and proliferation. Furthermore, metabolomic analysis showed that suppression of Ehdpck gene expression also caused decrease in the level of acetyl-CoA, and metabolites involved in amino acid, glycogen, hexosamine, nucleic acid metabolisms, chitin, and polyamine biosynthesis. The kinetic properties of E. histolytica and human DPCK showed remarkable differences, e.g., the Km values of E. histolytica and human DPCK were 58–114 and 5.2 μM toward dephospho-CoA and 15–20 and 192 μM for ATP, respectively. Phylogenetic analysis also supported the uniqueness of the amebic enzyme compared to the human counterpart. These biochemical, evolutionary features, and physiological importance of EhDPCKs indicate that EhDPCK represents the rational target for the development of anti-amebic agents

Eksplorasi Keanekaragaman Aktinomisetes Tanah Ternate Sebagai Sumber Antibiotik

Exploration of Soil Actinomycetes Diversity from Ternate as Indigenous Antibiotic Sources.Actinomycetes of soil samples from Ternate, North Moluccas were isolated using SDS-YEmethod in humic acid vitamin agar. Ternate has high abundance of Actinomycetes, approximately6.00 – 487 x 104 CFU/ g soil, depends on habitat types. We have selected 60 isolates andconducted antibiotic screening against pathogenic bacteria and fungi using agar diffusionmethod and found both narrow and broad antibiotic spectrum types . Based on 16S rDNAanalysis, all Actinomycetes with antibiotic activities are belong to the genus Streptomyces. .Minimum Inhibitor Concentration (MIC) value was determined by broth microdilution method.It was found that MIC values varied, depended on microbial tested. We found two isolateswith higher antibiotic activity compared to the commercial antibiotics (chloramphenicol,erythromycin for antibacterial and nystatin, kabicidin for antifungal). Cell destruction analysiscaused antibiotic activities was conducted through leak of protein and nuclatic acid

TOKSISITAS AKUT ORAL DUA SENYAWA BISANTRAKUINON (+)-2,2’-EPISITOSKIRIN A DAN (+)-1,1’-BISLUNATIN [Oral Acute Toxicity of Two Bisanthraquinones (+)-2,2’-Epicytoskyrin A and (+)-1,1’-Bislunatin]

Bisanthraquinones (+) - 2,2'-epicytoskyrin A and (+) -1,1'bislunatin produced by the endophytic fungus Diaporthe sp. GNBP-10 showed potent antibacterial activity on in-vitro test and have the opportunity to become new antibiotics candidates. The aspects of safety and toxicity of drug candidates have to be examined before applying to human. This study was conducted to determine the safety aspects of the compounds through acute oral toxicity testing in mice (Mus musculus). Acute toxicity of (+) - 2,2'-epicytoskyrin A and (+) - 1,1'-bislunatin evaluated by the method of Up and Down Procedure with limit test at a dose of 2000 mg / kg. Results of acute toxicity test showed that the LD50 of (+) - 2,2'-epicytoskyrin A and (+) - 1,1'-bislunatin were of 1638.87 mg / kg and > 2000 mg / kg respectively. Administration of (+)- 2,2'-epicytoskyrin A resulted in increased miliari multifocal hepatitis, fatty degeneration and necrosis of liver cells, and the renal tubule epithelial degeneration. Administration of (+) - 1,1'-bislunatin at a dose of 2000 mg / kg resulted in multifocal accumulation of inflammatory cells in the liver and degeneration of cells in the islets of Langerhans although not resulting in death. The administration of those compounds indicated the changes in the organs, but based on the UN/ECE classification of LD50 value showed that (+) 2,2'- epicytoskyrin A and (+) -1,1'-bislunatin included as low acute toxicity substance

Characterization and validation of Entamoeba histolytica pantothenate kinase as a novel anti-amebic drug target

The Coenzyme A (CoA), as a cofactor involved in >100 metabolic reactions, is essential to the basic biochemistry of life. Here, we investigated the CoA biosynthetic pathway of Entamoeba histolytica (E. histolytica), an enteric protozoan parasite responsible for human amebiasis. We identified four key enzymes involved in the CoA pathway: pantothenate kinase (PanK, EC 2.7.1.33), bifunctional phosphopantothenate-cysteine ligase/decarboxylase (PPCS-PPCDC), phosphopantetheine adenylyltransferase (PPAT) and dephospho-CoA kinase (DPCK). Cytosolic enzyme PanK, was selected for further biochemical, genetic, and phylogenetic characterization. Since E. histolytica PanK (EhPanK) is physiologically important and sufficiently divergent from its human orthologs, this enzyme represents an attractive target for the development of novel anti-amebic chemotherapies. Epigenetic gene silencing of PanK resulted in a significant reduction of PanK activity, intracellular CoA concentrations, and growth retardation in vitro, reinforcing the importance of this gene in E. histolytica. Furthermore, we screened the Kitasato Natural Products Library for inhibitors of recombinant EhPanK, and identified 14 such compounds. One compound demonstrated moderate inhibition of PanK activity and cell growth at a low concentration, as well as differential toxicity towards E. histolytica and human cells