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The Production of Nata Colored by Monascus purpureus J1 Pigments as Functional Food TRESNAWATI PURWADARIA; LISRINA GUNAWAN; AGUSTIN WYDIA GUNAWAN
Microbiology Indonesia Vol. 4 No. 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1055.427 KB) | DOI: 10.5454/mi.4.1.2

Abstract

Pigments from Monascus sp. may color nata. The mold also produces monacolin K that inhibits HMG-CoA reductase affecting the reduction of blood cholesterol. The aim of this study was to produce the colored nata containing monacolin K. The mold was isolated from commercial angkak. Potato sucrose (PS) and synthetic glucose (SG) media were used to ferment nata with Monascus purpureus J1. Fermentation of nata in PS medium produced red nata, while that in SG medium produced orange nata. The color of nata was similar to the color of supernatant. The optimum red production was obtained after five days of incubation, while the orange production increased until the 14th day. The color concentrations of the supernatant of PS medium containing nata (35.4 μg mL-1) were lower than those without nata (12.4 μg mL-1). The colors of nata looked darker than the color of the supernatant. The concentration of monacolin K in the red nata and the supernatant of PS medium were 0.6 μg mL-1 and 4.6 μg mL-1 respectively, while those in the orange nata and the supernatant of SG medium were 3.2 μg mL-1 and 14.6 μg mL-1 respectively. Dry matter biomass in the PS medium was lower than that in the SG medium. Even though the color of nata looked relatively stable, analyses of the nata water extract that showed a stable condition only occurred in freezing (-20OC) and soaking in buffer solution of pH 12; boiling, water washing, and soaking in a solution of pH 3 and 7 reduced the pigment concentration. Monacolin K concentration was not stable for every treatment, especially for water washing and freezing. Eventhough it was not stable, the boiling nata contained red pigments and monacolin K of 19.7 μg mL-1 and 0.1 μg mL-1 respectively, which can be served as functional food.
Isolation of Endophytic Bacteria from Palm Oil Fruits and Characterization of Their Lipases FANDY DJAFAR; TRESNAWATI PURWADARIA; ARNOLD PARLINDUNGAN SINURAT
Microbiology Indonesia Vol. 4 No. 2 (2010): August 2010
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (8305.143 KB) | DOI: 10.5454/mi.4.2.4

Abstract

Lipases (EC 3.1.1.3) can be produced in palm oil fruits by fruit cells or endophytic microbes. The purpose of this research is to isolate endophytic lipolytic bacteria of palm oil fruit and characterization their lipases. The bacteria were isolated and screened using medium Kouker and Jaeger containing olive oil, minerals, yeast extract, peptone, agar, and rhodamine B as an indicator. Fifteen endophytic bacteria were isolated and identified having the microbial lipase activity. Most of them showed rod shape, positively Gram test, spore formation, and motility except one bacteria strain K which was coccus. The enzyme was produced using submerged culture in the same medium but not containing agar and rhodamine B. Based on data of enzyme activity towards p-nitrophenyl palmitate as a substrate, protein concentration, and specific activity, two bacteria were selected, those were BSWt2(1) and Ink 1.3 isolates. Microscopic and biochemistry analyses show that BSWt2(1) and Ink 1.3 were identified as Bacillus brevis and B. lacterosporus respectively. Crude lipase from B. brevis BSWt2(1) and B. lacterosporus Ink 1.3 showed optimum activities at pH 8.0-9.0 and 60°C, and at pH 8.5 and 60-70°C. The enzymes were stable pre-incubated at pH 7.5-9.0 and pH 7.5-8.0 respectively, and they were stable pre-incubated for 2-4 hours at 80°C and for 2-8 hours at 100°C. Based on stability in high temperature, lipase from both isolates were specific and might be applicable for use in waste water treatment in palm oil factories.
GENETIC DIVERSITY ANALYSIS OF THERMOPHILIC BACTERIA FROM CANDRADIMUKA CRATER IN CENTRAL JAVA EMPLOYING PCR-RFLP OF 16S-rRNA GENE DESILIYARNI, TEMMY; SuwANTOAAAo, ANTONIUS; SUHARTONO, MAGGY T.; PURWADARIA, TRESNAWATI
BIOTROPIA No. 14 (1999)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (196.046 KB) | DOI: 10.11598/btb.1999.0.14.155

Abstract

The specific primers for bacteria (63f and 1387r) were used to amplify the 16S-rRNA genes from total community genomic DNA of thermophilic bacteria. The total community genomic DNA was obtained from muds and water samples of Candradimuka crater, Dieng Plateau, Central Java. PCR products were cloned into vector  pCR*2.1-TOPO (3.9 kb) and transformed into Escherichia coli TOPIC. Two  tetrameric restriction endonucleases  Rsal  and  Hhal  were employed to generate Restriction Fragment Length Polymorphisms (RFLP) paterns. These enzymes yielded 10 and 9 groups of 16S-rRNA profiles or OTU (Operational Taxonomic Units) from 27 16S-rRNA gene clones. Rsal was found to be more discriminative in differentiating the clones than Hhal. Rsal-RFLP indicated that OTU 7 and OTU 3 represented the most abundant clones, i.e. 6 and 5 clones respectively. The distribution of 16S-rRNA gene clones could  indicate relative distribution of  specific groups of  thermophilic  bacteria  in  their  natural habitat. Analysis of diversity at the DNA level could represent both culturable  and  unculturable bacteria in the  environment. Similarity analysis showed that  at level  0.600 there  were 8 different  groups from 10  RFLP  profiles generated by  Rsal  digestion. This study indicated that there were at least 8 groups of different thermophilic bacteria occupying Candradimuka crater. Key words: Thermophiles, 16S-rRNA, Candradimuka crater.
SYNERGISTIC ACTIVITY OF ENZYMES PRODUCED BY EUPENICILLIUM JAVANICUM AND ASPERGILLUS NIGER NRRL 337 ON PALM OIL FACTORY WASTES PURWADARIA, TRESNAWATI; NIRWANA, NONI; KETAREN, Pius P.; PRADONO, DYAH ISWANTINI; WIDYASTUTI, YANTYATI
BIOTROPIA No. 20 (2003)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2003.0.20.178

Abstract

The use of palm kernel cake (PKC) and palm oil mill effluent (POME), substances from palm oil factory wastes, for monogastric is limited by their high cellulose and mannan contents. Hydrolytic enzymes have been supplemented to increase the nutrient digestibility. The maximal digestibility was obtained in the synergistic action of all enzyme components including B-D-endoglucanase (CMCase), B-D-glucosidase, B-D-mannanase, p-D-mannosidase, and oc-D-galactosidase. Two kinds of enzymes produced by Eupenicillium javanicum  and Aspergillus niger NRRL 337 on the submerged culture containing 3% coconut meal were selected to hydrolyze PKC or dry POME. Enzyme from E. javanicum contained higher CMCase, B-D-mannanase, and a-D-galactosidase activities, while that from  A. niger  NRRL 337 contained more p-D-glucosidase and p-D-mannosidase activities. Saccharification (hydrolytic) activities of enzyme mixtures on PKC and POME were determined at pH 5.0, the optimal pH for p-D-mannanase from  E. javanicum,  and at 5.4 the optimal pH for a-D-galactosidase from  E. javanicum  and P-D-glucosidase from  A. niger NRRL 337. The enzyme proportions of E. javanicum and A. niger NRRL 337 were 100 : 0, 80 : 20, 60 : 40, 40 : 60, and 0 : 100%. The highest Saccharification activity on both substrates was observed on the mixture of 80% A. niger NRRL 337. The pH levels did not significantly affect Saccharification activity. Fiber components in PKC were more digestable than in POME. Further analysis on the reducing sugar components using thin layer chromatography showed that more monomers were produced in the 60 or 80% of A. niger NRRL 337. The glycosidases of A. niger NRRL 337 played more important role in the Saccharification activity. Keywords:        Synergistic activity/ palm kernel cake/palm oil mill effluent/ Eupenicillium javanicum/ Aspergillus niger NRRL 337
OPTIMIZATION OF CELLULASE PRODUCTION WITH PENICILLIUM NALGIOVENSE Sll GROWN ON PRETREATED WHEAT POLLARD PURWADARIA, TRESNAWATI; KUMALASARI, AGNES T.; TUTIHARYATI; KETAREN, PIUS P.; SINURAT1, ARNOLD P.
BIOTROPIA No. 23 (2004)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (360.752 KB) | DOI: 10.11598/btb.2004.0.23.202

Abstract

The cellulase production with Penicillium nalgiovense S11 on wheat pollard was enhanced using substrate pretreatments, i.e.: (i) mechanic process by Wiley milling, (ii) reducing sugars removal by water soaking, and (iii) chemical pretrcatment by 0.5% NaOH soaking at 100°C. The enzyme production stated as enzyme activities of all prctrcated substrates were higher than the untreated substrate. Although soaking with water showed significant increase in enzyme activities, the highest CMCase (EC 3.2.1.4), FPase (filter papcrase) and p-glucosidase (EC 3.2.1.21) were observed on NaOH pretreated pollard. The NaOH prctreatment also enhanced the enzyme production by increasing substrate concentration from 2 to 4%. The optimal incubation time in the cellulase production on 4% NaOH-pretrcated pollard was observed on the fifth day. Addition of 250 ppm glucose also increased the enzyme activities. The optimal treatments increased the specific activities of CMCase, FPase, and |3-glucosidase into 60, 4, and 198  times, respectively, as compared to the specific activities on 2% unpretreated pollard. Keywords  :   Cellulase production/ pollard pretreatments/ water soaking/ NaOH  soaking