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Improvement of Endoglucanase Activity in Penicillium oxalicum ID10-T065 Mutated by Ultra Violet Irradiation and Ethidium Bromide Caniago, Asnany; Mangunwardoyo, Wibowo; Nuswantara, Sukma; Lisdiyanti, Puspita
ANNALES BOGORIENSES Vol 19, No 2 (2015): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Penicillium sp. is known as filamentous fungi that produce complete cellulase. Cellulase. This study aims to improve endoglucanase activity of Penicillium oxalicum ID010-T065 by mutated with ultra violet irradiation (with dose of 0.1 J/cm2, 15 cm), ethidium bromide (10 Âµg/mL, 1 hour) and combination of both mutagens. The endoglucanase activity of all mutants was higher than that of the wild type (1.03 U/mL). Mutant UVEB-42 exposed to combine mutation showed the highest endoglucanase activity (2.76 U/mL) with a 2.70 fold increase. Mutant EB-45 (1.83 U/mL) exposed to ethidium bromide solution showed a 1.8 fold increase. Mutant UV-13 (1.72 U/mL) exposed to UV irradiation for 3 minutes showed a 1.7 fold increase. All mutants have optimum endoglucanase activity at 50 Â°C. Mutant UVEB-53 showed the highest thermostability by retaining 86 % of endoglucanase activity at 90 Â°C. The gene analysis of the endoglucanase I gene showed 3 bases mutated at mutant UV-13 and UVEB-53 that changed proline to serine. Mutant EB-45 showed 4 bases mutated that changed valine to glysine and proline to serine. Two bases mutated at Mutant UVEB-53 changed proline to serine. Bases mutated in eg1 gene could influenced the enhance of enzym activity in mutant.

Enhancement of Î²-Glucosidase Activity in Penicillium sp. by Random Mutation with Ultraviolet and Ethyl Methyl Sulfonate Syafriana, Vilya; Nuswantara, Sukma; Mangunwardoyo, Wibowo; Lisdiyanti, Puspita
ANNALES BOGORIENSES Vol 18, No 2 (2014): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

The genus Penicillium has a potential ability to produce Î²-glucosidase. The aim of the study was to improve the Î²-glucosidase activity of Penicillium sp. ID10-T065 with physical (Ultraviolet = UV), chemical (Ethyl Methyl Sulfonate = EMS), and combined mutation (UV-EMS). The spores of Penicillium sp. ID10-T065 were exposed into UV irradiation for 3 minutes with dose of 0.1 J/cm2 and 13 cm of distances. Chemical mutation was done by treated spores into 3% of EMS solution for an hour. Combined mutation of UV and EMS were also performed by UV for 3 minutes (0.1 J/cm2, 15 cm) and continued with soaking into 2-3% of EMS solution. The developed mutants were screened, selected and assayed. Comparison of enzyme activities with the wild- type (1.78 U/ml), mutant UV13 (5.53 U/ml) showed a 3.1 fold increase; mutant EM31 (4.26 U/ml) showed a 2.4 fold increase. Meanwhile, mutant UM23 obtained from the multiple exposures showed a decreased activity (1.75 U/ml). Mutant UV13 showed the best enzyme activity to be considered as a potential strain for Î²-glucosidase producer. This result needs to be further elaborated especially on its genetic stability studies in order for the ascertained as a stable mutant.

Conservation of Major L1 and Variability of Minor L2 Capsid Late Protein Genes in Human Papillomavirus of Indonesia Variants Nuswantara, Sukma; Prana, Titik K; Wulandari, Dwi; Widyowati, Henni; Anzela, Vera; Levy, Dea; Cahyadi, Petrus; Tjandra, Lukas D
ANNALES BOGORIENSES Vol 14, No 2 (2010): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Human Papilloma VirusÂ (HPV) has an out standing feature for its vast intraspecies variability. Of all known 100 types orÂ more, 15 t ypes of them are classified as high riskÂ because of their occurrenceÂ inÂ more than 95% of cervical cancer cases. Among all genes in their genome, E6 and E7 genes are considered oncogenes and have close relevance with their pathogenicity, whilst L1 and L2 ge nes produce capsid proteins that directly interact with theirÂ hostÂ receptors.Â ConsideringÂ theÂ importanceÂ ofÂ L1Â andÂ L2Â inÂ host-receptorÂ relationship,Â weÂ triedÂ to investigate their molecular variability thereby uncover their specificity as Indonesian variant s. Here we reported about the conservation of L1 minor capsid protein and variability of L2 capsid protein among high-risk types Human Papilloma Virus (HPV). The results indicated that L1 DNA was relatively more conserved than its L2 counterpart Also it was indicated that the middle part of either L1 or L2 CDSâ showed more DNA variability than thoseÂ atÂ theirÂ upstreamÂ sequences.Â ItÂ isÂ concludedÂ thatÂ L2Â middleÂ sequencesÂ areÂ importantÂ factorsÂ for intraspecific variations found in HPV of Indonesian variants. Â Keywords: Human Papilloma Virus (HPV), L1 gene, L2 gene, cervical cancer, high risk HPV

Efek Sumber Karbon Berbeda terhadap Produksi â-Glukan oleh Saccharomyces Cerevisiae pada Fermentor Air Lift Kusmiati, Kusmiati; Thontowi, Ahmad; Nuswantara, Sukma
Jurnal Natur Indonesia Vol 13, No 2 (2011)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

The need of â-glucan is increasing in food, medicine and cosmetic industry, because it becomes anticancer,antitumor and antiaging, increases immunosystem, and decreases cholesterol content in blood. The cell walls ofS. cerevisiae contain 80-90% polysaccharides that posses â-glucan. This research was aimed to obtain appropriatecarbon sources to increase the production of â-glucan. The carbon sources used were glucose, glucose commercial,sucrose and molases. The fermentation process was done by using air lift fermentor. The steps of fermentatonincluded regeneration of S. cerevisiae strain, preculture, fermentor preparation and running fermentor for 84hours. Sampling of S. cerevisiae culture was determined the cell growth by optical density (OD) usingspectrophotometer UV/VIS at ë 550 nm. The protein content was determined by Lowry method at ë 755 nm and thetotal glucose was measured by phenol sulphate method at ë 490 nm. The measurement result of cell growthshowed that the high intensity of S. cerevisiae in medium contain molases, but it did not show significant effectwhen compare to other carbon sources. The protein and carbohydrate contain in medium tended to decrease. Theresult of â-glucan on glucose, sucrose, glucose commercial and molases were 933,3, 1100, 1000, and966,7 mg/l. It can be concluded that sucrose and glucose commercial can replace the glucose to produce of â-glucan, because they are cheaper and easier to get. Beside that, molases can be used as an alternative carbonsource because it can produce of â-glucan as well as glucose.

POTENSI GEN ONCOPROTEIN HUMAN PAPILLOMAVIRUS TIPE 16 SEBAGAI KANDIDAT VAKSIN KANKER SERVIKS Taupiqurrohman, Opik; Yusuf, Muhammad; Nuswantara, Sukma; Subroto, Toto
Majalah Kedokteran Bandung Vol 48, No 2 (2016)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Kanker serviks menduduki peringkat dua besar penyebab kematian pada wanita dengan jumlah penderita meningkat setiap tahunnya. Kanker ini banyak diakibatkan oleh infeksi human papillomavirus (HPV) dan tipe prevalensi terbesar di Indonesia adalah tipe HPV 16. Vaksin HPV telah dikembangkan dan diproduksi secara komersial, namun perlu dicari alternatif lain vaksin dengan basis gen penyandi E (early) protein. Gen E6 dan E7 merupakan onkogen penting pada proses terjadinya kanker serviks. Penelitian ini dilaksanakan di Laboratorium Kimia Komputasi dan Bioinformatika, Universitas Padjadjaran, dari bulan Desember 2015 sampai Februari 2016. Pada studi ini, penemuan kandidat vaksin peptida HPV dilakukan dengan pendekatan imunoinformatika. Analisis in silico HPV tipe 16 menunjukkan bahwa gen E7 merupakan sumber kandidat vaksin yang potensial karena tidak homolog dengan genom manusia dan diprediksi memiliki afinitas yang baik terhadap major histocompability complex (MHC). Hasil analisis in silico menunjukkan bahwa kandidat vaksin HPV dari gen E7 adalah peptida YMLDLQPET dan HVDIRTLEDLLMGTL. [MKB. 2016;48(2):84?91]Kata kunci: HPV tipe 16, imunoinformatika, in silico, kanker serviksHuman Papillomavirus Type 16 Oncoprotein Genes as the Candidate of Cervical Cancer VaccineCervical cancer is the second largest cause of death for Indonesian women, with increasing number of cases every year. This cancer is mostly caused by human papillomavirus (HPV) infection, in which HPV 16 is the most prevalent type in Indonesia. Although HPV vaccine has been developed and commercially available, the other alternative of vaccine based on E (early) gene is required. Genes of E6 and E7 are important oncogenes in the development of cervical cancer. This study was conducted at the Laboratory of Computational Chemistry and Bioinformatics, Universitas Padjadjaran, from December 2015 to February 2016. In this study, the candidates of HPV peptide vaccine were discovered using immunoinformatics method. In silico-analysis of HPV type 16, it was shown gene E7 is not homologous with human genome and it is predicted to have a good affinity with major histocompability complex (MHC). Hence, it was proposed as a potential source of peptide vaccine. It is concluded that he candidates forHPV vaccine from E7 peptides are YMLDLQPET and HVDIRTLEDLLMGTL. [MKB. 2016;48(2):84?91]Key words: Cervical cancer HPV 16, immunoinformatics, in silico

PRODUKSI BETA-1,3 GLUKAN DARI Agrobacterium DAN AKTIVITAS PENYEMBUHAN LUKA TERBUKA PADA TIKUS PUTIH Kusmiati, Kusmiati; Rachmawati, Fitria; Siregar, Syafrida; Nuswantara, Sukma; Malik, Amarila
Makara Journal of Science Vol. 10, No. 1
Publisher : UI Scholars Hub

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Production of beta-1,3 glucan from Agrobacterium and its wound healing activity on white rat. The objective of this study was to determine the activity of beta-1,3 glucan product extracted from local Agrobacterium sp Bro 1.2.1, both wild-type and mutant-type, on opened-wound healing process. Beta-1,3 glucan product was extracted by precipitation, and the purification was carried out by column chromatography as KCl gradient fractions. In this study, white Sprague Dawley rats were employed, and have been treated for opened-wound condition. Seven groups were performed in this experiment, i.e. the negative control, the positive control employing povidone iodine, the two groups of two commercial beta-1,3 glucan with 0,02 mg/4 cm2 each, and the last three groups of beta-1,3 glucan as the test group with 0,02 mg/4 cm2, 0,10 mg/4 cm2 and 0,50 mg/4 cm2, respectively. The result showed significant differences of wound-healing activity performing statistical analysis of the least significance between the negative control, the positive control, as well as the highest dose of the test group of beta-1,3 glucan, at the dose of 0,5 mg/4 cm2 (p<0.05).

PRODUKSI BETA-1,3 GLUKAN DARI Agrobacterium DAN AKTIVITAS PENYEMBUHAN LUKA TERBUKA PADA TIKUS PUTIH Kusmiati, Kusmiati; Rachmawati, Fitria; Siregar, Syafrida; Nuswantara, Sukma; Malik, Amarila
Makara Journal of Science Vol. 10, No. 1
Publisher : UI Scholars Hub

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Production of beta-1,3 glucan from Agrobacterium and its wound healing activity on white rat. The objective of this study was to determine the activity of beta-1,3 glucan product extracted from local Agrobacterium sp Bro 1.2.1, both wild-type and mutant-type, on opened-wound healing process. Beta-1,3 glucan product was extracted by precipitation, and the purification was carried out by column chromatography as KCl gradient fractions. In this study, white Sprague Dawley rats were employed, and have been treated for opened-wound condition. Seven groups were performed in this experiment, i.e. the negative control, the positive control employing povidone iodine, the two groups of two commercial beta-1,3 glucan with 0,02 mg/4 cm2 each, and the last three groups of beta-1,3 glucan as the test group with 0,02 mg/4 cm2, 0,10 mg/4 cm2 and 0,50 mg/4 cm2, respectively. The result showed significant differences of wound-healing activity performing statistical analysis of the least significance between the negative control, the positive control, as well as the highest dose of the test group of beta-1,3 glucan, at the dose of 0,5 mg/4 cm2 (p<0.05).

PRODUKSI BETA-1,3 GLUKAN DARI Agrobacterium DAN AKTIVITAS PENYEMBUHAN LUKA TERBUKA PADA TIKUS PUTIH Kusmiati, Kusmiati; Rachmawati, Fitria; Siregar, Syafrida; Nuswantara, Sukma; Malik, Amarila
Makara Journal of Science Vol. 10, No. 1
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Production of beta-1,3 glucan from Agrobacterium and its wound healing activity on white rat. The objective of this study was to determine the activity of beta-1,3 glucan product extracted from local Agrobacterium sp Bro 1.2.1, both wild-type and mutant-type, on opened-wound healing process. Beta-1,3 glucan product was extracted by precipitation, and the purification was carried out by column chromatography as KCl gradient fractions. In this study, white Sprague Dawley rats were employed, and have been treated for opened-wound condition. Seven groups were performed in this experiment, i.e. the negative control, the positive control employing povidone iodine, the two groups of two commercial beta-1,3 glucan with 0,02 mg/4 cm2 each, and the last three groups of beta-1,3 glucan as the test group with 0,02 mg/4 cm2, 0,10 mg/4 cm2 and 0,50 mg/4 cm2, respectively. The result showed significant differences of wound-healing activity performing statistical analysis of the least significance between the negative control, the positive control, as well as the highest dose of the test group of beta-1,3 glucan, at the dose of 0,5 mg/4 cm2 (p<0.05).

PENGARUH PENAMBAHAN URASIL DALAM MEDIA FERMENTASI TERHADAP HASIL β-GLUKAN DARI DUA GALUR AGROBACTERIUM Kusmiati, Kusmiati; Tamat, Swasono R.; Nuswantara, Sukma; Muhamad, Salmah
Makara Journal of Science Vol. 11, No. 2
Publisher : UI Scholars Hub

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Influence of Uracil in Fermentation Media on β-Glucan Production by Agrobacterium Radiobacter A 1.5 and Agrobacterium sp. Bro 1.2.1. Optimum β-glucan production can be achieved by an optimum condition in the fermentation media. Uracil, as a precursor of UDP-glucose, may act as a glucose donor in the formation of polysaccharides such as β-glucan. It is expected that addition of certain quantity of uracil into the fermentation media in a suitable growth phase of Agrobacterium radiobacter A 1.5 and Agrobacterium sp. Bro 1.2.1, will significantly increase the β-glucan production. In this investigation, 0.025%; 0.05% or 0.1% of uracil were added into the fermentation media during the logarithmic phase (24 hour) or stationary phase (46 hour) of growth. The β-glucan product was evaluated from the β-glucan (crude) dry-weight and from the β-glucan content. Beta-glucan content was determined as glucose by the Hisamatsu-AOAC and HPLC methods. The highest β-glucan (crude) dry-weight produced by the A. 1.5 was in a medium containg 0.025% uracil (24 hour), whilst by the A. Bro 1.2.1 was in a medium containg 0.1% uracil (46 hour), both higher than control. The highest β-glucan content produced by the A. 1.5 (27.03%) was in a medium containg 0.025% uracil (46 hour), while control produced only 23.28%. The highest β-glucan content produced by the Bro 1.2.1 (29.34%) was in a medium containg 0.025% uracil (24 hour), while control produced only 28.75%. Two-way anova analysis showed that there were no significant influence difference (α = 0,05) from various concentration of uracil in either growth phases, to the yield of β-glucan (crude) dry-weight nor to the β-glucan equivalent glucose content.

Beta-Glucosidase 1 (bgl1) Gene Analysis in Mutant and Wild-type of Penicillium sp. ID10-T065 Syafriana, Vilya; Nuswantara, Sukma; Mangunwardoyo, Wibowo; Lisdiyanti, Puspita
Jurnal Riset Biologi dan Aplikasinya Vol. 4 No. 1 (2022)
Publisher : Universitas Negeri Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26740/jrba.v4n1.p1-8

In the previous study, Penicillium sp. ID10-T065 has been mutated using ultraviolet (UV), ethyl methyl sulfonate (EMS) and the combination of UV-EMS to increase Beta-glucosidase (bgl) activity. There were three mutants selected, UV13 (UV mutant), EM31 (EMS mutant), and UM23 (UV-EMS mutant). This study examined the mutations in the bgl gene encoding (bgl1) as well as sequence differences between mutants and wild-type of Penicillium sp. ID10-T065. The gene analysis was performed by PCR amplification and sequencing of the bgl1 gene. The results of bgl1 gene sequences (600 bp) from mutants were aligned with the wild-type, it was discovered that there were base alterations from position 2025 to 2050. Mutant UV13 showed the highest base alterations (7 bases) which occurred at position 2027 (T?C); 2036 (T?G); 2040 (T?G); 2047 (G?C); and 2048-2050 (TTG?GGA). Mutant EM31 showed alterations in five bases at positions 2034 (G?A), 2036 (T?G), 2037 (G?C), 2044 (G?C), and 2047 (G?T). Mutant UM23 showed two base alterations at position 2025 (T?A) and 2037 (G?C). UV irradiation and EMS mutation resulted in transition and transversion DNA, whereas the combination of UV-EMS mutation resulted in transversion mutations. Base alterations in UV13 and EM31 mutants, causing missense and silent mutation, while in UM23 mutant only silent mutations occur. The bgl1 gene analysis showed that mutation using UV light was more effective than using EMS or a combination of UV-EMS in Penicillium sp. ID10-T065.

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