cover
Filter by Year
Articles by issue : Vol 1, No 2 (2007): August 2007
11
Articles
Products of Orange II Biodegradation by Enterococcus faecalis ID6017 and Chryseobacterium indologenes ID6016

MEITINIARTI, VINCENTIA IRENE ( Universitas Gadjah Mada ) , SOETARTO, ENDANG SUTARININGSIH ( Universitas Gadjah Mada ) , TIMOTIUS, KRIS HERAWAN, SUGIHARTO, EKO

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (261.229 KB)

Abstract

Chryseobacterium indologenes and Enterococcus faecalis were isolated from activated sludge of textile wastewater treatment plant. These bacteria had the ability to decolorize several azo-dyes. Degradation of azo dyes was initiated by decolorization (reduction of azo bond) which occurred in anaerobic condition. In this study, we focussed on biodegradation of Orange II by pure culture of C. indologenes ID6016 and E. faecalis ID6017, and to determine the metabolite products of Orange II degradation. The degradation of Orange II by both bacteria was carried out in batch experiments using liquid medium containing 80 mg/l Orange II, under sequential static agitated incubation. During the bacterial growth under static incubation (6 h), 66.1 mg/l Orange II were decolorized by 35.54 mg/l biomass of E. faecalis ID6017, but no decolorization found with C. indologenes ID6016. Based on HPLC results, the decolorized Orange II products were identified as sulfanilic acid and amino-naphthol. These metabolites were probably used or degraded by C. indologenes ID6016 under agitated incubation.

Evidence for a Link Between Pathogenicity and the Role of ImpBacterial Transport Effector Proteins in Soybean Infection by Xanthomonas axonopodis pv. glycines

SUWANTO, ANTONIUS ( Bogor Agricultural University ) , WAHYUDI, ARIS TRI ( Bogor Agricultural University ) , TJAHJONO, BUDI ( Bogor Agricultural University )

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (970.126 KB)

Abstract

Xanthomonas axonopodis pv. glycines (Xag) is the causal agent of bacterial pustule disease of soybeans. A nonpathogenic mutant of Xag (M715) was constructed employing transposon mutagenesis which showed similar epiphytic survival in planta to its wild type strain (YR32). The objective of this work was to identify and to analyze genes involved in pathogenicity in Xag YR32. Inverse Polymerase Chain Reaction (IPCR) was used to isolate the DNA flanking transposon insertion. A 1.3 kb flanking DNA fragment was sequenced and analyzed employing BLAST program to study homology, the position of transposon insertion and to predict the structure and function of the gene. One of the Open Reading Frames (ORFs) shared homology with inner membrane proteins (imps) of Xanthomonas axonopodis pv. citri (GenBank accession No. NC 003919). Northern blot analysis revealed that an imps gene was monocistronic and the size of imps mRNA in YR32 was slightly longer than in M715. Reverse Transcriptase- PCR analysis demonstrated that the imps transcript in M715 was much less abundant than in the wild type YR32. Transposon (mini-Tn5-Kmr-Tpr) was determined to be inserted close to the end of C-terminal region of imps gene and might be sufficient to destabilize the imps transcript in M715 and so influence effectors transportation from Xag to plant cell.

Formation and Pathogenicity Variation of Oospores of Phytophthora capsici Infecting Black Pepper

MANOHARA, DYAH ( Indonesian Research Institute for Medicinal and Aromatic Crops, Pusat Penelitian dan Pengembangan Perkebunan, )

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (435.656 KB)

Abstract

Phytophthora capsici Leonian is the causal agent of foot rot disease of black pepper (Piper nigrum L.). Foot rot disease is the most destructive disease which can cause significant economic losses of black pepper. Two mating types of P. capsici were found in black pepper plantations in Lampung. This research was aimed at examining the effects of temperature, light, and polycarbonate membrane on oospore formation. Also the effect of light on germination and determining both mating types of 30 randomly selected progenies. The results showed that oospores were produced through either hyphal contact or without any contact. Oospores were produced abundantly on V8 agar in the dark at 16-24 oC. Oospores germinated after 3-weeks incubation in the dark followed by 1-week incubation under TL fluorescent light. All progeny were heterothallic, and consisted of 16 isolates of A1 mating type and 14 isolates of A2 mating type. There was no correlation between mating type categories and their pathogenicities on black pepper leaves. Twenty-six progeny isolates may infect unwounded pepper leaves, whereas another four isolates may only infect wounded pepper leaves. Pathogenicities of all progenies were lower than those of their parents.

The Dynamics of Bacterial Communities During Traditional Nata de Coco Fermentation

SEUMAHU, CECILIA ANNA, SUWANTO, ANTONIUS, HADISUSANTO, DEBORA, SUHARTONO, MAGGY THENAWIJAYA

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (582.948 KB)

Abstract

One of the important problems in traditional Nata de Coco (Nata) fermentation is production inconsistency due to strain or genetic variability reflecting mixed microbial communities involved in this process. This research was aimed at examine the population dynamics of the bacterial community during the fermentation processes. Samples were collected daily for six days from fermentation media derived from “good” and “bad” Nata fermentation. We compared the levels of bacterial diversity through amplified 16S-rRNA (ARDRA). DNA was extracted directly from the fermentation media and 16S-rRNA gene was amplified employing Universal Bacterial Primers. The amplicons were cloned into pGEM-T Easy vector, and restriction enzymes HaeIII and RsaI were used to generate ARDRA profiles. ARDRA phylotypes of DNA extracted from the fermentation medium obtained from different Nata qualities were compared. Phylotype profiles demonstrated unique bacterial community profiles for different conditions of Nata quality, which could be developed as a parameter to monitor Nata quality during fermentation. In this research we found that the dynamics of the bacterial population involved in Nata fermentation were a crucial factor for determining traditional Nata quality.

Screening of Proteolytic Enzymes of Streptomyces sp. Local Strain and Their Characterization

YURATMOKO, DERI, MUBARIK, NISA RACHMANIA, MERYANDINI, ANJA

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (236.721 KB)

Abstract

Protease of two Streptomyces sp. strain were chosen for characterization because of the large clear zone surrounding the colony in nutrient agar media containing 1% (w/v) skim milk. Extracellular protease from the two isolates SLW 8-1 and 45I-3 were characterized following incubation of the isolate in Nutrient Broth media containing skim milk or chicken feather (1%). The optimum activity of the protease SLW 8-1 was at pH 9 and 80 ºC, whereas that of the keratinase was at pH 6.5 and 70 +C. Protease of strain 45I-3 showed its optimum activity at pH 7.5 and 50 ºC whereas the keratinase was at pH 8.5 and 80 ºC.

The Process of Xylanase Production from Bacillus pumilus RXAIII-5

RICHANA, NUR, IRAWADI, TUN TEDJA, NUR, M. ANWAR, SAILAH, ILLAH, SYAMSU, KHASWAR

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (324.944 KB)

Abstract

The optimum conditions for the growth of Bacillus pumilus RXAIII-5 (a potential xylanase producer) were sought, these included temperature, pH, aeration, and agitation of the culture batch. Afterwards a mathematical model based on the parameter of cultivation kinetics was formulated. At the same time, the rheology of the fluid used for bacterial cultivation in a bioreactor was studied. The data obtained was used for estimating the ‘scaling up’ of enzyme production. The results of the study indicate that the optimum condition for processing in 50 ml Erlenmeyer flask are used temperature of 35 oC (308oK), pH 7, and an agitation rate of 140 rpm. The highest xylanase activity and its specific activity are 297.132 U.ml-1 and 655.32 U.g-1protein, respectively. Subsequent experiments in a bioreactor using all of the experiment parameters mentioned above, except for the agitation rate, shows that the results are as follows. The highest specific growth was at 0.082 hour-1 at an aeration and agitation rate of 0.5 vvm and 150 rpm, respectively. Based on the data of the cultivation kinetics, the optimum conditions for the fermentation in Biostat 2L-bioreactor is 1 vvm and 200 rpm of aeration and agitation, respectively . The efficiency of substrate (Yp/s) and of cell biomass (Yp/x) to produce xylanase is 50.744 U.g-1 and 43.906 U.g-1, respectively. The efficiency of substrate to cell production (Yx/s) is 1.178g.g-1. The liquid cultivation-medium has non-Newtonian properties. Based on a mathematical model it is found that the consistency index (k constant) and index of liquid behavior (n value) are 0.179 g.cm-1.second-1 and 0.3212, respectively. Becouse the value of 0

Characterization of Acid-Aluminium Sensitive Mutants of Soybean Symbiont Bradyrhizobium japonicum Generated by Transposon Mutagenesis

WAHYUDI, ARIS TRI, PURNAWIJAYA, ANDINI, NURDIANI, DINI, IMAS, TEDJA

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (61.648 KB)

Abstract

Acid-aluminium sensitive mutants of symbiotic bacterium Bradyrhizobium japonicum BJ11 (designated as AAS11) and KDR15 (designated as AAS15) were constructed by mini-Tn5 transposon mutagenesis to study genes involved in acid-aluminium tolerance (AAT) in B. japonicum. Transposon delivery was carried out through conjugation between B. japonicum strains as recipients and Escherichia coli S17-1 (ë pir) carrying pUTmini- Tn5Km1 as a donor strain. The result showed that frequency of transconjugation was in the range of 6.7 x 10-7 to 7.1 x 10-6 cell per recipients. AAS11 and AAS15 mutants did not grow on Ayanaba media (pH 4.5) containing 50 μM Aluminium. These mutants remained able to form root nodules of Siratro (Macroptilium arthropurpureum) plants revealing genes interrupted by transposon which were responsible for acid-Al tolerance did not correlate with the nodulation genes. Strains tolerant to acid-aluminium and their mutants with a wild type sensitive to acidaluminium were characterized by accumulating phosphate and aluminium absorption. Compared to the wild type acid-aluminium tolerant B. japonicum, there was approximately a three- to eight-times decrease in phosphate accumulation and a five- to seven-times increase in aluminium absorption by these mutants. These results suggest that aluminium and phosphate contents in the bacterial cells may be involved in mechanisms of acid-Al tolerance of B. japonicum grown in acid-aluminium stress conditions.

Generation and Characterization of Temperature Resistant Mutant of Recombinant PJ156/CAV-17 Virus

UTAMA, ANDI, SHIMIZU, HIROYUKI

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (467.677 KB)

Abstract

Previous study revealed that a recombinant virus between poliovirus (isolate PJ156) and coxsackie A virus serotype 17 (CAV-17), namely PJ156/CAV-17, was temperature sensitive. It is well known that two amino acids in 3D region (His-73 and Ile-362) are determinants for temperature sensitivity of poliovirus, in particular for Sabin 1 strain. However, it is not known whether those amino acids affect the temperature sensitivity for other enteroviruses. Sequence analysis of 3D region of PJ156/CAV-17 showed that amino acid in 3D-73 and -362 were Tyr and Ile, respectively, similar with the sequence of parental CAV-17 virus. Since amino acid in 3D-73 of PJ156/ CAV-17 was not His, it is suggested that the temperature sensitivity of the PJ156/CAV-17 was associated with the Ile-362. To confirm this suggestion, the temperature-sensitive escape mutants of PJ156/CAV-17 were generated by blind passaging at 39.5 oC. The escape mutants were then recovered and plaque purified, and the sequence of 3D region was determined. It was found that the amino acid in 3D-362 was Thr, instead of Ile. Consequently, Ile- 362 was proved to be involved in temperature sensitivity of PJ156/CAV-17. Full sequences of both viruses were also determined and compared. Furthermore, the characteristics of the temperature resistant of PJ156/CAV-17 variant were analyzed. It is confirmed that the recovered PJ156/CAV-17 virus could grow well at 39.5 oC, and there was strong correlation between temperature sensitivity and attenuation.

Antiphatogenic and Anti Food Spoilage Activities of Ethylacetate and Methanol Extract of Panax ginseng var. Notoginseng

YASNI, SEDARNAWATI

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (45.598 KB)

Abstract

Javanese ginseng is a traditional herb known to possess broad health benefits that have been clinically proven. The aim of this research was to analyze the antimicrobial activity of Javanese ginseng against pathogenic bacteria (Escherichia coli, Salmonella typhimurium, Bacillus cereus, Staphylococcus aureus), food spoilage bacteria (Bacillus stearothermophilus and Pseudomonas fluorescens) and food spoilage fungi (Aspergillus flavus, Fusarium graminearum, and Penicillium citrinum). The result may increase the utilization of ginseng not only for health purposes but also as a natural food preservative. It may also open new possibilities for the development of natural functional foods. Ethylacetate and methanol extracts, obtained by maceration, were fractionated employing vacuum liquid chromatography (VLC). Fractionation using methanol and ethylacetate as solvents produced six fractions from each solvent. Fractions 1 and 4 of methanol extract performed the highest growth inhibitory effects on Bacillus cereus (Gram-positive bacteria) and Escherichia coli (Gram-negative bacteria), whereas fractions 4 and fraction 5 of methanol extract effectively inhibited the growth of Penicillium citrinum.

The Production of Tannin Acyl Hydrolase from Aspergillus niger

ANWAR, YUNITA ARIAN SANI, HASIM, ., ARTIKA, I MADE

Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

Show Abstract | Original Source | Check in Google Scholar | Full PDF (56.212 KB)

Abstract

The aim of this research was to produce tannin acylhydrolase (tannase) from Aspergillus niger isolated from cacao pod. The first step of the study included determination of optimal pH, temperature, and incubation period to produce tannase. Optimal conditions obtained for tannase production were pH 5.5, a temperature of 28 oC and an incubation period of 3 days. Optimization of production medium was conducted. The media tested were solid and liquid wheat flour media with a concentration of tannic acid as inducer at 0, 3, 5, and 7% (wt/vol). The best production medium was solid medium with tannic acid concentration of 5% (wt/vol).