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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 : BRIN

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Abstract

Human Papilloma Virus (HPV) has an outstanding feature for its vast intraspecies variability. Of all known 100 types or more, 15 types of them are classified as high risk because of their occurrence in 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 genes produce capsid proteins that directly interact with their host receptors. Considering the importance of L1 and L2 in host-receptor relationship, we tried to investigate their molecular variability thereby uncover their specificity as Indonesian variants. 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 CDS‟ showed more DNA variability than those at their upstream sequences. It is concluded that L2 middle sequences are important factors for intraspecific variations found in HPV of Indonesian variants.
Enhancement of β-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 : BRIN

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The genus Penicillium has a potential ability to produce β-glucosidase. The aim of the study was to improve the β-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 β-glucosidase producer. This result needs to be further elaborated especially on its genetic stability studies in order for the ascertained as a stable mutant.
Improvement of Endoglucanase Activity in Penicillium oxalicum ID10-T065 Mutated by Ultra Violet Irradiation and Ethidium Bromide Caniago, Asnany; Mangunwidjaja, Wibowo; Nuswantara, Sukma; Lisdiyanti, Puspita
Annales Bogorienses Vol. 19 No. 2 (2015): Annales Bogorienses
Publisher : BRIN

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Abstract

Penicillium sp. is known as a filamentous fungus that produces complete cellulase. This study aims to improve endoglucanase activity of Penicillium oxalicum ID010-T065 by mutation with ultra violet irradiation (with dose of 0.1 J/cm2, 15 cm), ethidium bromide (10 μg/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 °C. Mutant UVEB-53 showed the highest thermostability by retaining 86 % of endoglucanase activity at 90 °C. The gene analysis of the endoglucanase I gene (eg1) showed 3 bases mutation in mutant UV-13 and UVEB-53 that changed proline to serine. Mutant EB-45 showed 4 mutated bases that changed valine to glysine and proline to serine. Two bases mutation in Mutant UVEB-53 changed proline to serine. Bases mutation which is occured in eg1 gene could influence the enhance of endoglucanase activity in mutant.