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Massive In Vitro Propagation Of Sandalwood Through Friable Embryogenic Callus

Sandalwood (Santalum album), which belongs to Santalaceae family, is a commercially important tree in Indonesia due to its many application. However,its population has significantly depleted since the planting materials using conventional methods are difficult to be provided. This study was conducted to mass propagate sandalwood using in vitro methods through friable embryogenic callus (FEC). The somatic embryos were formed using leaves cultured in MS +0.5 mg/l +1 mg/l indole acetic acid (IAA), and MS +1 mg/l IAA + 0.2 mg/l kinetin as well as 0.5 MS+1 mg/l Gibberellic acid (GA3). Primary somatic embryos (PSE) and secondary somatic embryos (SSE) then formed friable embryogenic callus when they were repetitively transferred to MS +1.5 mg/l BAP + 1.2 mg/l kinetin every 3 weeks. The maturation and regeneration of FEC was best done in the MS +1.5 mg/l BAP + 1.2 mg/l kinetin for 4-8 weeks. The acclimatization of sandalwood plantlets can be best conducted in the medium containing soil, sand and compos in ratio of 1:1:1 with the companion plant Murraya paniculata, which gave the best percentage of survival rate and the lowest percentage of falling leaves.

 

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Expression of no Affinity Tagged Recombinant Human Interferon Alpha-2a in Methilotropic Yeast Pichia pastoris

Recombinant human interferon alpha-2a (rhIFNα-2a) has been widely used for clinical therapy as antiviral, anticancer as well as immunomodulator. In this study, The open reading frame (ORF) encoding synthetic hIFNα-2a was constructed to be in framed with N-terminal alpha factor secretion system in methylotropic yeast Pichiapastoris. This research covered construction of no affinity tagged recombinant human interferon alpha-2a, expression and analysis of the molecular weight and identity of the protein. We used pPICZαB plasmid for cloning and expression vector. The confirmed recombinant plasmid containing the correct DNA sequence of hIFNα-2a was linearized by SacI restriction enzyme and then transformed into P. pastorisgenome using electroporation. We screened two multi-copy recombinants in YPDS plates containing zeocin™. Buffered complex medium containing methanol (BMMY) was used for protein expression during 48 hours in the culture condition. The recombinant protein was purified by blue sepharose affinity chromatography. Analyses of protein hIFNα-2a by SDS-PAGE and Western blot confirmed that protein band observed at around 19.2 kDa was recombinant hIFNα-2a. The quantification of purified rhIFNα-2a by using colorimetric binichoninic assay (BCA) informed that the yield was 44 mg/L culture (OD600= 2-3).

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Antiplasmodial activity of methanol extract and pure isolated compound of Calophyllum bicolor P. F. Steven

Calophyllum bicolor (Clusiacea) is a big tree from Indonesian rain forest in Palangkaraya, Kalimantan. Calophyllum or bintagor is one of many sources of natural bioactive compounds that can be used in the fields of health and pharrmaceuticals. The aim of this research was to explore the ability of methanol extract of Calophyllum bicolor P.F. Steven to inhibit the growth of Plasmodium falciparum. The methanol extract was purified by colomn chromatographyc system, hexane – ethyl acetate was used as solvent with increasing polarity. One pure compound was obtained and was elucidated based on the 1D and 2D-NMR, COSY DEPT, HMBC and HMQC data and the isolated compound was identified as xanthone. Methanol extract showed growth inhibition against parasite P. falciparum with IC50 5.2 ppm and pure compound have maximum inhibition at concentration 0.11 nMol respectively

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Characterization of Phytase, Amylase and Cellulase by Aspergillus niger, Neurospora crassa and Rhizophus oryzae on Sargassum and Rice bran under Solid State Fermentation

The objective of study was to characterize phytase, amylase, and cellulase on sargassum and rice bran on solid state fermentation using Aspergillus niger, Neurospora crassa and Rhizophus oryzae. Media for solid state fermentations composed of dried sargassum and rice bran. The effect of particle of sargassum, initial moisture content on phytase, amylase, and cellulase were evaluated. Optimum enzyme activity of phytase, amylase and cellulase were obtained after 4 days fermentation at 30 °C, and initial moisture content was adjusted to 60 %. The optimum particle size of dried sargassum attaining the highest enzyme activity was 25 mesh. Best formula for enzymes production was at the ratio of 4:6 (w/w) of Sargassum spinosum (SS) and rice bran (RB) respectively. At this formula highest phytase activity was obtained by A. niger, amylase by N. crassa, and cellulase by R. oryzae. Media composed of sargassum and rice bran can be used for phytase, amylase and cellulase production.

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Improvement of Endoglucanase Activity in Penicillium oxalicum ID10-T065 Mutated by Ultra Violet Irradiation and Ethidium Bromide

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 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 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.

 

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