cover
Filter by Year
Articles by issue : Vol 6, No 2 (2012): June 2012
5
Articles
The Effect of Growth Medium pH towards Trypsin-Like Activity Produced by Lactic Acid Bacteria

WULANSARI, DYAH, WAHYUNTARI, BUDIASIH, TRISMILAH, TRISMILAH, NURHASANAH, ASTUTIATI

Microbiology Indonesia Vol 6, No 2 (2012): June 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (1337.584 KB)

Abstract

In cases of pancreatic disease, trypsin deficiency often occurs due to reduced expression of trypsin in the pancreas. Patients with pancreatic problem can be treated with a supplement containing digestive enzymes, including trypsin. However, most of the enzymes currently used for the treatment are derived from porcine and bovine sources. On the other hand, lactic acid bacteria are also known to show trypsin-like activity. In the previous work, our group screened 11 lactic acid bacteria isolates, which had previously been proven to show serine protease activity, for trypsin-like activity. The strains were initially grown in MRS (de Mann, Rogosa and Sharpe) medium before being transferred directly to the production medium to produce trypsin. During the previous study, the initial pH of the production medium was set at 6 (the same as the MRS medium pH), which is the optimum pH for the cell growth of lactic acid bacteria. However, most trypsin has an optimum pH of around 8. In this study, we altered the production medium pH to 8 and we harvested the lactic acid bacteria from MRS medium by centrifugation prior to their inoculation to the production medium. Observation of the culture growth and enzyme activity indicated that the new strategy improved the enzyme activity expressed by some strains.

Properties of an Extracellular Protease of Bacillus megaterium DSM 319 as Depilating Aid of Hides

WAHYUNTARI, BUDIASIH ( BPPT ) , HENDRAWATI, HENDRAWATI ( UIN )

Microbiology Indonesia Vol 6, No 2 (2012): June 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (1929.409 KB)

Abstract

Properties of a Bacillus megaterium DSM319 extracellular protease which are related to its application for depilating hides were investigated. The properties observed were optimum temperature and pH, the type of protease, and type of the protein which could be hydrolyzed by the enzyme. The enzyme was produced in a 3.5 liters LKB jar fermentor in a medium containing (2.0% v v-1 molasses and 1.3% w v-1 urea at 37 ºC , pH 7.5, aeration 1 vvm, agitation 250 rpm for 24 hours). The enzyme solution was concentrated by means of membrane ultrafiltration followed by ammonium sulfate precipitation at 70% w v-1 saturation. Optimal temperature and pH were observed at 30 ºC and pH 8 respectively. Two mM PMSF and EDTA inhibited the enzyme leaving level of activity of 84.5 and 4.3% correspondingly, indicated that the crude enzyme might be a metal requiring serine protease. The presence of 0.5, 2.0, and 3.5 mM CaCl caused an increase of the enzyme activity of 73, 88, and 79% respectively. The enzyme was able to hydrolyze Na-Benzoyl-DL-Arginine p-Nitro Anilide, a specific amino acid sequence cleaved by trypsin at a reaction rate of 0.024 absorbance value at 405 nm per minute. The enzyme was capable of hydrolyzing bovine serum albumin, hemoglobin, and gelatin, and to hydrolyze alkaline soluble collagen and keratin. The K value of the enzyme for hydrolysis of bovine serum albumin and gelatin was 3.44 m and 1.65 mg mL-1, whereas V values were 8.09 and 55.24 mg mL-1 respectively. The experimental data showed max that the crude enzymes have potential for dehairing of cowhides.

Five Unique Amino Acid Residues of Hemagglutinin (HA) Proteins of Swine Influenza A (H1N1) Detected in 2009 in Jakarta, Indonesia

YASMON, ANDI ( Department of Microbiology, Faculty of Medicine University of Indonesia, Jl. Pegangsaan Timur 16 Jakarta 10320 Indonesia ) , MUHAYAR, YULIANTY ( Department of Microbiology, Faculty of Medicine University of Indonesia, Jl. Pegangsaan Timur 16 Jakarta 10320 Indonesia ) , SETIAWATY, VIVI ( National Institute of Health Research and Development, Ministry of Health, Jakarta, Indonesia ) , DEWI, BETI ERNAWATI ( Department of Microbiology, Faculty of Medicine University of Indonesia, Jl. Pegangsaan Timur 16 Jakarta 10320 Indonesia ) , BELA, BUDIMAN ( Department of Microbiology, Faculty of Medicine University of Indonesia, Jl. Pegangsaan Timur 16 Jakarta 10320 Indonesia ) , IBRAHIM, FERA ( Department of Microbiology, Faculty of Medicine University of Indonesia, Jl. Pegangsaan Timur 16 Jakarta 10320 Indonesia )

Microbiology Indonesia Vol 6, No 2 (2012): June 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (976.836 KB)

Abstract

Nine HA genes of influenza A (H1N1) viruses originating from swine which were detected in 2009 in Jakarta, Indonesia, were characterized in this study. Nasopharyngeal and/or pharyngeal samples were extracted to obtain viral RNA genomes. Amplification of the HA segment was performed by using the reverse transcription-polymerase chain reaction (RT-PCR), and followed by nested PCR in cases of RT-PCR negative. DNA sequencing was performed by using eight overlapping primers. All the Jakarta strains were closely related to vaccine strain A/California/07/2009. Nine amino acid changes were found in the Jakarta strains, and 5 (P100S, S220T, G239D, R240Q, and I338V) of those were unique to all Jakarta strains with respect to strain A/California/07/2009 used to produce vaccine. An I338V substitution was detected in a cleavage site of HA and no amino acid changes were detected in potential sites for N-linked glycosylation. For seven sites (positions 131, 158, 160, 183, 187, 222, and 227) playing an important role in viral attachment to host receptor, none of the expected amino acid changes was detected; however, a S220T substitution close to amino acid 222 was found in all the Jakarta strains. All amino acid changes potentially affect the pathogenicity of the viruses and the efficacy of strain A/California/07/2009 in neutralizing the Jakarta strains.

Isolation and Identification of a Thermostable Amylase-Producing Bacterium from Hatuasa Hotspring

MALLE, DOMINGGUS ( Faculty of Agriculture, Pattimura University, Jalan Ir M Putuhena, Kampus Poka Ambon, Ambon 97233, Indonesia ) , PICARIMA, JUNUS ( Faculty of Mathematics and Natural Sciences, Pattimura University, Jalan Ir M Putuhena, Kampus Poka Ambon, Ambon 97233, Indonesia ) , HUWAE, LAURY CHARA ( Faculty of Mathematics and Natural Sciences, Pattimura University, Jalan Ir M Putuhena, Kampus Poka Ambon, Ambon 97233, Indonesia ) , RAHMAWATI, INDRA ( Center for Biotechnology , BPPT, Puspiptek-Serpong, Tangerang Selatan, Indonesia ) , PURBOWASITO, WAHYU ( Center for Biotechnology , BPPT, Puspiptek-Serpong, Tangerang Selatan, Indonesia )

Microbiology Indonesia Vol 6, No 2 (2012): June 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (1801.672 KB)

Abstract

Hot springs are a common source of thermophyles which produce thermostable enzymes. The objective of this study was to isolate and identify thermostable amylase-producing bacteria from a local geothermal spring. An amylase–producing bacterium strain was isolated from this hot spring which excreted amylase after being grown on starch agar screening plates at 37 °C. It was identified as Bacillus megaterium using the method of 16S ribosomal DNA. The organism is a rod-shape and is a spore-forming  bacterium.  Maximum amylase production was achieved after incubation in the production media for 72 hours. Preliminary analysis of the secreted amylase showed that the enzyme could bind to DEAE-Sepharose matrix and was discharged by eluting with 0.5 M NaCl. The partially purified enzyme was stable up to 75 °C, showing that this enzyme might have potential application in the starch-processing industry.

Mercury (Hg)-Resistant Bacteria in Hg-Polluted Gold Mine Sites of Bandung, West Java Province, Indonesia

CHAERUN, SITI KHODIJAH ( Laboratory of Biogeosciences, Mining and Environmental Bioengineering, Research Division of Genetics and Molecular Biotechnology, School of Life Sciences and Technology, Institut Teknologi Bandung, Jalan Ganesha 10, Bandung 40132, Indonesia Center ) , HASNI, SAKINAH ( Laboratory of Biogeosciences, Mining and Environmental Bioengineering, Research Division of Genetics and Molecular Biotechnology, School of Life Sciences and Technology, Institut Teknologi Bandung, Jalan Ganesha 10, Bandung 40132, Indonesia ) , SANWANI, EDY ( Department of Metallurgical Engineering, Faculty of Mining and Petroleum Engineering, Institut Teknologi Bandung, Jalan Ganesha 10, Bandung 40132, Indonesia ) , MOEIS, MAELITA RAMDANI ( Research Division of Genetics and Molecular Biotechnology, School of Life Sciences and Technology, Institut Teknologi Bandung, Jalan Ganesha 10, Bandung 40132, Indonesia )

Microbiology Indonesia Vol 6, No 2 (2012): June 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (1762.508 KB)

Abstract

In the present study, ten mercury-resistant heterotrophic bacterial strains were isolated from mercurycontaminated gold mine sites in Bandung, West Java Province, Indonesia. The bacteria (designated strains SKCSH1- SKCSH10) were capable of growing well at ~200 ppm of HgCl except for strain SKCSH8, which was able to grow at 550 ppm HgCl . The bacteria were mesophylic and grew optimally at 1% NaCl at neutral pH with the optimal growth temperature of 25-37 ºC. Phenotypic characterization and phylogenetic analysis based on the 16S rRNA gene sequence indicated that the isolates were closely related to the family Xanthomonadaceae, Aeromonadaceae, and Pseudomonadaceae and they were identified as Pseudomonas spp., Stenotrophomonas sp., and Aeromonas sp. Eight bacterial strains were shown to belong to the Pseudomonas branch, one strain to the Stenotrophomonas branch and one strain to the Aeromonas branch of the ã-Proteobacteria. Phylogeny based on their 16S rRNA gene sequences indicated that four of the isolates (SKCSH1, SKCSH4, SKCSH7, SKCSH9) could be classified as representatives of four novel species in the genus Pseudomonas that were allocated to P. moraviensis (96.96% similarity) and P. plecogossicida (94.53, 96.61, and 96.73% similarity). Four other isolates could be allocated to P. plecogossicida (97.57 and 98.66% similarity) and P. hibiscicola (99.97% similarity), one isolate to Stenotrophomonas africana (99.69% similarity), and one other isolate to Aeromonas hydrophila subsp. ranae (99.43% similarity). The findings of this study provide the first information of the phylogenetically-diverse Hg-resistant bacteria in the Hg-polluted sites of Indonesia that may be highly useful for developing in situ bioremediation or detoxification of Hg-contaminated sites in Indonesia.