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Peroksidasi Lipid oleh Parasetamol dan Ekstrak Air Panas Teh Hijau (Camellia sisnensis ) pada Sel Khamir Candida tropicalis yang di Simpan pada Suhu Rendah Julistiono, Heddy
JURNAL BIOLOGI INDONESIA Vol 8, No 1 (2012): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v8i1.3070

ABSTRACTLipid Peroxidation by Paracetamol and Hot Water Extract of Green Tea (Camellia sinensis )Â in Yeast Candida tropicalis preserved in Low Temperature. The use of C. tropicalis cell as aÂ tool to evaluate antioxidant property of green tea to protect oxidative stress caused byÂ paracetamol in cell level was investigated in our laboratory. Immediate availability of cellÂ culture will significantly reduce the time of cell preparation. Low temperature preservation ofÂ cell culture is one of methods to produce cell cultures. However, low temperature might affectphysiological state of the cell. In this study, effect of low temperature preservation (4 ÂºC, 10Â days) on the oxidative response of yeast cell treated with paracetamol and hot water extract ofÂ green tea had been investigated. Cells incubation with paracetamol 0.3 % for 2 h causedÂ oxidative stress in both fresh and preserved culture since the content of a marker of oxidativestress, peroxyd lipids increased significantly. Whereas, concentration of peroxidised lipids inÂ preserved cultures was lower than that of fresh culture. Increasing of peroxydized lipids followedÂ with decreasing of cell viability in fresh culture but not in preserved culture. Green tea withconcentration of 0.1 % decreased peroxyd lipids in fresh cultures treated with paracetamol butÂ not in preserved cultures. Green tea with concentration of 0.2 % or 0.4 % in increased peroxydÂ lipids in fresh cultures treated with paracetamol but not in preserved cultures. The data indicatedÂ that green tea did not show anti- or pro-oxidative effect in cultures preserved in low temperatureÂ treated with paracetamol. However, induction of super oxide dismutase (SOD), an antioxidantÂ defense enzyme, was not observed in cell preserved in low temperature. The study revealedthat low temperature might make the cell more resistant against prooxidative properties ofÂ paracetamol.Keywords: C. tropicalis, oxidative stress, low temperature, paracetamol, green tea

Eksplorasi Keanekaragaman Aktinomisetes Tanah Ternate Sebagai Sumber Antibiotik Nurkanto, Arif; Listyaningsih, Febrianti; Julistiono, Heddy; Agusta, Andria
JURNAL BIOLOGI INDONESIA Vol 6, No 3 (2010): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

ABSTRACTExploration 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 â 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.Key words : Actinomycetes, soil, Ternate, antibiotic, cell distruction

Induction of Superoxide Dismutase Activities and Ethanol Resistance by Sodium Chloride and Lead Acetate in Yeast Candida sp. Y390 Julistiono, Heddy
JURNAL BIOLOGI INDONESIA Vol 4, No 1 (2006): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v4i1.3271

ABSTRAKAktivitas Induksi Superoksida Dismutase dan Resistensi Etanol oleh Sodium Kloridadan Plumbum Asetat pada Khamir Candida sp. Y390. Untuk mengetahui sifat khamirCandida sp. Y390 dalam melindungi diri dari cekaman oksidatif, telah dilakukan risetkajian toleransi sel terhadap etanol dan hubungannya dengan enzim superoksida dismutase(SOD) pada khamir tersebut. Viabilitas sel yang ditumbuhkan pada media gliserol dandiinkubasi pada etanol 17,s % selama 1 jam adalah 0.30 f 0.09 % , pada media gliseroldan 1 1,7 % natrium khlorida adalah 1.65 f 0.5 % sedang pada media giserol dan 1 ppmplumbum asetat adalah 1.16 f 0.7 %. Aktivitas CuZnSOD pada sel yang diinduksi olehnatrium khlorida atau plumbum asetat turun sedikit, yakni masing-masing 1,7 dan 1,9kali. Aktivitas MnSOD pada sel yang diperlakukan dengan natrium khlorida naik sedikit(sekitar 1,03 kali), namun pada sel yang diperlakukan dengan plumbum asetat naik secaramenyolok (sekitar 3,6 kali). Data ini menunjukkan bahwa metabolisme etanol dapatmenyebabkan Candida sp. Y390 mengalami cekaman oksidatif. MnSOD khamir mungkinberperan dalam melindungi sel dari kerusakan akibat cekaman oksidatif.Keywords: Candida sp. Y390, yeast, ethanol metabolism, oxidative stress, SOD.

Superoxide Dismutase Activity and Ethanol Respiration in a Fungi Resistance to Ethanol Monascus sp. MM Julistiono, Heddy; Suharna, Nandang; Desnora, Beni
JURNAL BIOLOGI INDONESIA Vol 3, No 4 (2002): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

ABSTRACTMonascus sp. MM was a contaminant fungus isolated from museum specimenpreserved with ethanol 70 %. In order to verify role of superoxide dismutase (SOD) inprotecting cell from ethanol toxicity during ethanol metabolism, SOD activities of Monascussp.MM and a Monmcus sp. NGK, which was isolated fiom fermented red rice (angkak), werecompared. When fungus was grown with glucose, CuIZn-SOD activity of Monascus sp., MMwas 7.1 times of thht of Monascus sp. NGK. Whereas in ethanol medium, CuIZn-SOD activityof Monarcus sp. MM was 24.6 times of that in Monascus sp. NGK. Induction of CuIZn-SODMonmcus sp. MM by ethanol was not observed. Compared with Mn-SOD, activity of CuIZn-SOD was markedly important (I 0 times of Mn-SOD when fungi grown with ethanol; 12 timeswhen the fungi grown with glucose). The data indicated that Cu/Zn-SOD might play animportant role in protecting cell fiom ethanol toxicity during ethanol metabolism. Ethanolrespiration rate of Monascus sp. MM was also important since O2 consumption and ethanoldegradation rates were clearly higher than that of Monascus sp. NGK.Keywords: Monarcw sp., superoxide dismutase, respiration, ethanol resistance.

ANALISIS DELIMITASI JENIS PADA Monascus Spp. MENGGUNAKAN SIDIK JARI DNA ARBITRARY PRIMER PCR [Species Delimitation Analysis within Monascus spp. using Arbitrary Primer PCR DNA Fingerprinting] Suharna, Nandang; Julistiono, Heddy
BERITA BIOLOGI Vol 15, No 2 (2016)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v15i2.2928

A species delimitation analysis within Monascus spp. using Arbitrary Primer Polymerase Chain Reaction (AP PCR) DNA fingerprint was carried out. This is one of the methods used for identification and discrimination of bacterial strains within the same species. Its advantages including using single primer, independent of DNA quality, and observing amplicon shared by only some strain. This study analyzed Monascus sp. MM isolate which was originated from a source contaning high level of ethanol and two M. purpureus isolates which isolated from angkak. However, based on ITS region, 99% homology showed the unclear species delimitation. Therefore, this analysis was aimed at clarifying on the identities of Monascus species tested. The result showed DNA polymorphism among three isolates of Monascus that showed species delimitation. This study showed that species delimitation within Monascus isolates used in this analysis could be supported by AP PCR DNA fingerprinting. Therefor we suggested to use this technique or method for phylogenetic study to clarify taxonomic position of Monascus strains.Â

EFEK FERRI SITRAT TERHADAP KEMAMPUAN KHAMIR Candida tropicalis DALAM MEREDUKSI 3-(4,5-DIMETHYLTHIAZOL-2-YL)- 2,5-DIPHENYLTETRAZOLIUM BROMIDE (MTT) [Effect of ferric citrate on 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) reduction in yeast Candida tropicalis] Julistiono, Heddy; Muthmainah, Resti Siti; Adam, Adam
BERITA BIOLOGI Vol 11, No 2 (2012)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Effect of iron (ferric citrate) on 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) reduction in yeast Candida tropicalis was investigated. Reduction of MTT in yeast grown in YMB media containing 5 mM of ferric citrate decreased significantly compared to that of yeast grown containing 0, or 1.25 or 2.5 mM ferric citrate after 24 h incubation. However, there was no difference in cell density among cultures treated with 0 mM, 1.25 mM, and 5 mM ferric citrate. Ferric citrate of 5 mM caused smaller colony when cells were grown on YPDG media. Reduction of MTT in smaller colony cells was weaker than that of with normal size colony. An antioxidant, Epigallocatechin Gallate (EGCG) of 0.01 % could not reverse MTT reduction caused by 5 mM ferric citrate. Since enzymes responsible in MTT reduction are usually located in mitochondrion, the data suggested that, in the condition of 5 mM ferric citrate might cause mitochondrion disorder without killing the yeast cells. The data was in concordance with other studies on other yeast or human cells. However, this study does not show role of free radicals provoked by high level of iron concentration.

ANALISIS VARIASI GENETIK Saccharomyces cerevisiae Dl TAHAN ETANOL DENGAN RAPD (RANDOM AMPLIFIED POLYMORPHIC DNA) [Genetic Variation Analysis of Ethanol Tolerance Yeast, Saccharomyces cerevisiae Dl by Using RAPD] Julistiono, Heddy; Yulineri, Titin; Hanjono, Sukamto
BERITA BIOLOGI Vol 4, No 5 (1999)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Genetic variations among 3 cultures, which were treated with or without Mn of Saccharomvces cerevisiae D1, were analyzed using RAPD (Random Amplified Polymorphic DNA ) technique. The Mn-treatment of three cultures were as follows: the culture KMn was D1 strain, the culture Mn+ was D1 strain colony survived in ethanol 20%, which was previously treated with 0,5 mU MnSOt and the culture Mn- was a D1 colony survived in ethanol 20% without MnSOi treatment. Polymorphism of total DNA of the three cultures may indicate that mutation may occur in cells which were tolerant to ethanol. The locus or base change was not identified. However, since the oxygen uptake rate of the three cultures at catabolite derepression state were identical,the results suggest that the locus may not be in mitochondrial DNA encoding respiratory chain proteins. The relation between DNA polimorphic and ethanol tolerant cell is still to be clarified.

CEKAMAN OKSIDASI SEL KHAMIR Candida tropicalis YANG DIPERLAKUKAN DENGAN PARASETAMOL DAN ANTIOKSIDAN (+)-KATEKHIN [Oxidative Stress in Candida tropicalis Treated with Paracetamol and Antioxidant (+)-catechin] Julistiono, Heddy
BERITA BIOLOGI Vol 11, No 1 (2012)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v11i1.1889

In order to more understand similarity of yeast Candida tropicalis with mammalian cells in analgesic drug paracetamol metabolism and toxicity, ability of yeast in the drug metabolism and oxidative response of cells treated with the drug and (+)-catechin was studied. In mammalian cells, paracetamol toxicity is mainly caused by metabolite byproduct of drug metabolism catalyzed by cytochrome P450, a membrane-bound enzyme and peroxidase and a soluble enzyme. Previously it has been shown that paracetamol induced oxidative stress in the yeast cells; and green tea extract protected the cells from oxidation. In this study, it had been shown that paracetamol could be metabolized by yeast cell suspension or cell free extracellular protein, reflecting possibility of role of enzyme that can not be separated from cell and that is soluble, which is common phenomenon in mammalian cell system. Paracetamol of 3 mg/ml increased lipid peroxidation, a marker of oxidative stress. A green tea polyphenol, (+)-catechin of 0.1 mg/ml did not decrease lipid peroxidation content induced by paracetamol. At higher concentration (2 mg/ml), solely (+)-catechin did not increase lipid peroxidation content. Paracetamol or (+)-catechin induced slightly activity of superoxide dismutase enzyme. The data indicated that paracetamol metabolism or toxicity in the yeast may be similar to that of mammalian cells. In this condition, it suggested that (+)-catechin is one of polyphenol green tea that has weak activity of antioxidant and consequently has weak activity of prooxidant. Mechanism of paracetamol toxicity in C. tropicalis is still to be studied with emphasis on the free radical formation and drug metabolism.

Penguraian Parasetamol oleh Sel dan Protein Ekstraselular Khamir Candida tropicalis dan Rhodotorula minuta Julistiono, Heddy; Saragih, Ernawati; Yulineri, Titin
Media Penelitian dan Pengembangan Kesehatan Vol 27, No 3 (2017)
Publisher : Badan Penelitian dan Pengembangan Kesehatan

Yeast can be used as cell model to study toxicity in mammalian cell. In the previous study we demonstrated that yeast Candida tropicalis was able to metabolize analgesic drug paracetamol causing oxidative stress. This phenomenon is similar to that in mammalian cell. In mammalian cell system, enzymes responsible in paracetamol metabolism are at least cytochrome P450 (P450) and peroxidase. In order to understand the possible role of peroxidase enzyme in paracetamol metabolism in yeast, research on the effect of peroxidase inhibitor of sodium cyanide (KCN) and a peroxidase substrate peroxide (H2O2) on paracetamol degradation by cell suspension and extracellular protein of C. tropicalis and Rhodotorula minuta was carried out. Paracetamol was degraded by cells or extracellular protein in both of yeast. Paracetamol degradations were significantly inhibited by KCN (0.01 Î¼M) or H2O2 (3 Î¼M). Since P450 is generally located inside the cell (in cell membrane) while no activity of P450 in extracellular, the data indicated the presence of soluble enzyme which is able to metabolize paracetamol that is inhibited by KCN or H2O2. The possibility of presence of peroxidase in the soluble protein by which paracetamol is metabolized and its inhibition by peroxide via competitive substrate or peroxide toxicity is discussed. The results supported use of yeast for studying toxicity of paracetamol in cell level.AbstrakKhamir dapat digunakan sebagai sel model untuk mempelajari toksisitas pada sel mamalia. Pada penelitian sebelumnya diketahui bahwa khamir Candida tropicalis mampu memetabolisme obat analgesik parasetamol dan mengakibatkan terjadinya cekaman oksidasi pada sel seperti yang terjadi pada sel mamalia. Pada sel mamalia, metabolisme parasetamol terutama dilakukan setidaknya oleh enzim membran sitokrom P450 dan peroksidase. Untuk mengetahui indikasi keterlibatan enzim peroksidase dalam metabolisme parasetamol pada C. tropicalis dan Rhodotorula minuta, diamati efek senyawa inhibitor peroksidase kalium sianida (KCN) terhadap metabolisme parasetamol; juga efek hidrogen peroksida (H2O2), senyawa substrat peroksidase. Hasil menunjukkan bahwa pada kedua khamir, baik pada sel maupun protein ekstraselular dapat mengurai parasetamol. Penguraian parasetamol dapat dihambat oleh KCN (0,01 Î¼M) dan juga H2O2 (3 Î¼M). Mengingat pada umumnya khamir memiliki P450 dalam sel (membran) tetapi tidak ada aktivitas P450 pada larutan ekstraselular, maka hasil ini mengindikasikan adanya peran enzim terlarut dalam mengurai parasetamol, yang dihambat oleh KCN dan H2O2. Kemungkinan enzim terlarut tersebut adalah peroksida yang kemampuan metabolisme parasetamolnya dapat dihambat oleh H2O2 melalui proses kompetitif dan keracunan dibahas dalam penelitian ini. Kemampuan metabolisme parasetamol oleh enzim yang diduga peroksidase menambah peluang penggunaan khamir dalam penelitian toksisitas parasetamol di tingkat sel.

SIFAT PROTEKSI EKSTRAK AIR PANAS TEH {Camellia sinensis (L.) Kuntze} HIJAU PADA KHAMIR Candida tropicalis YANG DIPERLAKUKAN DENGAN PARACETAMOL Julistiono, Heddy
BERITA BIOLOGI Vol 10, No 6 (2011)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

In order to develop yeast Candida tropicalis as a model cell for evaluation of substance having anti- or pro-oxidant activity in cell level, the effect of hot water tea {Camellia sinensis (LJ Kuntze) extract on peroxidized lipids, a marker of oxidative stress and cell mortality in the yeast caused by paracetamol has been evaluated. Incubation of yeast cell in the presence of 1.38 % green tea for 2 h decreased cell viability followed with increasing of peroxidized lipids, whereas 0.27 % green tea caused increasing of peroxidized lipids without causing cell death.Yeast cell was not affected by 0.1 % green tea. Incubation of yeast cell with presence of 0.15 % (g/v) paracetamol for 2 h did not cause cell mortality, however content of peroxidized lipids increased significantly. In the presence of higher dose of paracetamol (0.3 %) cell viability remarkably decreased and followed with increasing of peroxidized lipids significantly. Green tea of 0.1 % increased cell viability of cells treated with 0.3 % paracetamol while peroxidised lipids decreased. The data indicated that high dose of paracetamol caused oxidative stress in yeast cells, while green tea with lower concentration might protect paracetamol toxicity due to its antioxidant property. Although the antioxidant property, high dose of green tea could cause oxidation due to its pro-oxidant activity. In conclusion, yeast C. tropicalis could be potentially used as a model cell to evaluate substances having antioxidant property in cell level.

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