S. Margino, S.
Fakultas Pertanian Universitas Gadjah Mada

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ISOLATION AND LIGNOCELLULOLYTIC ACTIVITIES OF FIBER-DIGESTING BACTERIA FROM DIGESTIVE TRACT OF TERMITE (Cryptothermes sp.)

Journal of the Indonesian Tropical Animal Agriculture Vol 39, No 4 (2014): December
Publisher : Diponegoro University

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Abstract

The objectives of this study were to obtain the fiber-digesting bacteria isolates from termitedigestive tract and to determine the optimum conditions of growth and production of cellulase, xylanaseand ligninase enzyme of isolate. The first study was conducted to isolate and select the fiber-digestingbacteria from the digestive tract of termites based on the highest activity of cellulolytic (S), xylanolytic(X) and lignolytic (L). The second study was optimation of the growth conditions of bacteria and theenzyme production due to effect of rice straw substrate and nitrogen. The material used were dry woodtermites, rice straw, and culture medium. The design used was a completely randomized factorial design,in which the first factor was rice straw substrate (1, 2, and 3% W/V), while the second factor wasnitrogen (0.1, 0.2 and 0.3% W/V). Variables measured were cellulase, xylanase and ligninase activities.Results of the first sudy showed that the isolates obtained consisted of 3 types, those were cellulolyticbacteria (S1, S2, and S3), 3 types of bacteria xylanolytic (X1, X2, and X3) and 3 types of bacteria lignolytic(L1, L2, and L3). Meanwhile, results of the second study showed that isolates of S2, X3, and L1 had thehighest activity, those were 1.894 U/mL, 1.722 U/mL and 0.314 U/mL, respectively. In conclusion, the addition of 1% level of rice straw substrate and 0.3% of nitrogen showed the highest enzyme activity oncellulase, xylanase and ligninase.

Isolasi dan Karakterisasi Β-1,3-Glukanase Akar Semai Tusam (Pinus merkusii Jungh. et de Vriese) yang Berasosiasi dengan Fungi Ektomikorisa

Jurnal Perlindungan Tanaman Indonesia Vol 11, No 2 (2005)
Publisher : Universitas Gadjah Mada

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Association between tusam (Pinus merkusii Jungh. et de Vriese) seedling roots with ectomycorrhiza fungi was expected to increase the activity {β-1,3-glucanase level in the plants. This enzyme has potency to protect seedling from soil borne fungal pathogens by degrading the fungal cell walls. The objectives of this research were to isolate and characterize β-1,3-glucanase from tusam seedling roots associated with ectomycorrhiza fungi. Crude protein was isolated with ammonium sulfat precipitation and then purified by gel filtration chromatography and characterize molecular weight, temperature and pH for optimum the activity. The enzyme from tusam seedling roots associated with ectomycorrhizal fungi, designated as GLUC15 have 15 kD molecular weight, 30° - 40° C temperature and pH 5-7 for optimum activity.

Pemurnian dan Karakterisasi Enzim Endokitinase dari Agen Pengendali Hayati Trichoderma reesei

Jurnal Perlindungan Tanaman Indonesia Vol 7, No 2 (2001)
Publisher : Universitas Gadjah Mada

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This experiment was aimed to purify and characterize the endochitinase of Trichoderma reesei. Extracellular endochitinase was produced by T. reesei strain T13, a fungal biocontrol agent in colloidal chitin medium as sole carbon source. The enzyme was purified by ammonium sulfate precipitation, followed by gel filtration chromatography and chromatofocusing. The results showed that T. reesei produced endochitinase with molecular weight of 32 kDa and the activity was optimum at pH of 5,5 and temperature of 30 to 35oC.Key words: endochitinase, Trichoderma reesei

Isolasi dan Karakterisasi Kitinase Akar Tusam (Pinus merkusii Jungh. et de Vriese) Roots yang Bersimbiosis dengan Fungi Ektomikoriza

Jurnal Perlindungan Tanaman Indonesia Vol 11, No 2 (2005)
Publisher : Universitas Gadjah Mada

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The experiment was aimed to detect the activity and characterize the chitinase of tusam root during ectomycorrhizal symbiosis. Tusam inoculated with tusam stands soil from Kaliurang as fungi inocula. Crude proteins were isolated from 4, 6, and 8 weeks age of tusam. Chitinase activity and isoform were detected using glycol chitin as a subtrat. The enzyme was purified by ammonium sulfat precipitation, dialysis, followed by gel filtration chromatography. The results showed that tusam prodced chitinasse with a molecular weight of approximately 52kDa. The optimum activity was at pH 5 and temperature of 30°C.

Skrining Bakteri Asam Laktat Penghasil Bakteriosin dari Daging dan Produk Olahannya

Agritech Vol 24, No 2 (2004)
Publisher : Faculty of Agricultural Technology, Universitas Gadjah Mada, Yogyakarta, Indonesia

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Lactic acid bacteria (LAB) which naturally occur in meat and meat products have been isolated and screened for their ability to produce bacteriocin. The objective of this research was to obtain the potential bacteriocin producer of lactic acid bacteria which could be used as food bio-preservative. Source of lactic acid bacteria used in this study were beef chicken flesh, vacuum packaged sausage and sliced meat obtained from traditional market or department store. Ten grams of each samples was put onto five different enrichment media, i.e., TGE (tryptoneglucose-yeast extract) pH 5 plus 3% NaCl; MRS (deMan Rogose Sharpe) pH 5,5; TGE broth pH 5,5; TGE buffer broth pH 5,5; and TGE broth plus Tween 80 & 1% Naazida pH 6,0, incubated for 24-71 hours to stimulate the growth of lactic acid bacteria. Different enrichment media were used to stimulate the growth of strains belong to each genus, since the nutritional and environmental requirement for optimum growth were suggested to be genera-dependent. Screening of LAB bacteriocin producer was carried out by dilution -pour plate methods (culture from each enrichment medium) followed by overlay using the indicator strains. Indicator strains used in this study were Lactobacillus plantarum NCDO 955, Pediococcus acidilactici LB-42, Leuconostoc mesenteroides LY, and Enterococcus faecalis MI. Colonies showing growth inhibition to indicator (indicated by clear zone) were isolated and purified. Isolates were then characterized based on Gram, catalase, shape and arrangement of cell, type of fermentation, effect of temperature to the growth and acid production from several carbon sources. From the primary screening (dilution - pour plate –overlay), 30 strains belong to Lactobacillus, Leuconostoc, Streptococcus and Enterococcus which suspected to produce antimicrobial substance were obtained. However, based on the confirmation lest (dUsion method), only three (3) strains were identified to produce bacteriocin. i. e. Leuconostoc mesenteroides SM 22, SM 32, and SM 46. In this study, Leuconostoc mesenteroides SM 22 was selected for food application. Bacteriocin of Leuconostoc mesenteroides SM-22 was able to inhibit the growth of psychrophilic bacteria naturally occur in meat and shrimp kept at refrigerator. Microbial population of raw meal with the initial number of about 3x104 CFU/g decreased one log cycle after treated with bacteriocin, and this number maintained less than 105 CFU/g after storage raw meat at refrigerator for five days. On the other side, microbial population of raw meat with no bacteriocin treatment increased to 106 CFU/g after 4 days kept at refrigerator. In the case of shrimp, washing raw shrimp with cold water could reduce the population of bacteria about one log cycle, followed treatment with bacteriocin, this populationincreased very slowly and still less than 105 CFU/g after 5 days storage at refrigerator. While without any treatment, microbial population of raw shrimp which initially about 3x105 CFU/g rapidly increased to 106 CFU/g after 3 days. This data showed that Leuconostoc mesenteroides SM-22 was a potential bacteriosin producer and can be applied as bio-preservative for cold storage fbod.

Transformasi Fragmen DNA Salmonella Ke Dalam Sel E. coli DH5-α

Agritech Vol 17, No 1 (1997)
Publisher : Faculty of Agricultural Technology, Universitas Gadjah Mada, Yogyakarta, Indonesia

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Genomic DNA from Salmonella typhimarium ATCC 14028, cultured in a Luria broth for 24 hr at 37°C, was isolated from the cells using EDTA as a chelating agent and SDS as a detergent and lysing agent. The amount of genomic DNA was then digested with each of three restriction endonucleases, i.e., EcoRI, HindlII, and BamHI for 4 hr at 37°C. A plasmid (pUC19) was used as a vector. Prior to use, the pUC19 plasmid was split with one of the three restriction enzymes and dephosphorylated using a bacterial alkaline phosphatase. The genomic DNA was then ligated to the corresponding prediggested plasmid using a ligase at 15°C over night. Transformation of the DNA recombinant into E.coli DH5-a was successfully carried out using a heat shock method as indicated by gel electrophoresis