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SELEKSI DAN IDENTIFIKASI BAKTERI ENDOFIT POTENSIAL PENGHASIL ENZIM PROTEASE DARI TAMAN NASIONAL GUNUNG HALIMUN - (The Selection and Identification of Potential Endophyte Bacteria as Protease Enzyme Producer from Halimun Mount National Park) Melliawati, Ruth; Rohmattusolihat, Rohmattusolihat; Nuryati, Nuryati; Rahmani, Nanik; Yopi, Yopi
Biopropal Industri Vol 7, No 2 (2016)
Publisher : Balai Riset dan Standardisasi Industri Pontianak

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (416.906 KB)

Endophytic bacteria have an equal chance to bacteria that live outside the plant tissue as potential bacteria. The selection has done towards 326 bacterial endophyte isolates. This research aimed to find and identify proteolytic potential isolates. The proteolytic selection of endophytic bacteria had done using solid skim milk. The capability of endophytic bacteria to agglomerate milk was tested using liquid skim milk which incubated for 7 days at room temperature. Enzyme production of four selected isolates was made through fermentation in GYS medium. The results showed that 86 isolates have proteolytic potential. Isolate HL.29B.63 had highest protease enzymes activity (65.918 U/mL). Medium optimization was able to increase the enzyme activity into 89.94% (125.04 U/mL). The analysis used 16s rDNA showed that isolate HL.29B.63 was Bacillus amyloliquefacient subs. plantarum strain FZB42.Keywords: endophytic bacteria, fermentation, identification, protease, selectionÂ ABSTRAKBakteri endofit mempunyai peluang yang sama dengan bakteri yang hidup diluar jaringan tanaman sebagai bakteri potensial. Seleksi dilakukan terhadap 326 isolat bakteri endofit. Tujuan penelitian ini adalah mencari isolat yang berpotensi proteolitik dan mengidentifikasinya. Seleksi proteolitik terhadap bakteri endofitik menggunakan skim milk padat. Uji kemampuan bakteri endofitik dalam menggumpalkan susu menggunakan medium skim milk cair yang diinkubasi selama 7 hari pada suhu ruang. Produksi enzim terhadap empat isolat terseleksi dilakukan melalui fermentasi dalam medium GYS. Hasilnya menunjukkan bahwa 86 isolat mempunyai potensi proteolitik. Isolat HL.29B.63 mempunyai aktif enzim protease tertinggi (65,918 U/mL). Optimasi medium dapat meningkatkan aktivitas enzim sebesar 89,94% (125,04 U/mL). Analisis menggunakan 16s rDNA menunjukkan bahwa isolat HL.29B.63 adalah Bacillus amyloliquefaciens subs. plantarum strain FZB42.Kata kunci: bakteri endofit, fermentasi, identifikasi, protease, seleksi

ANALISIS KARBOKSIMETIL SELULOSA DARI BAKTERI Acetobacter xylinum DAN Acetobacter sp. RMG-2 [Analysis of Carboxymethyl Cellulose from Acetobacter xylinum and Acetobacter sp. RMG-2 Bacteria] Melliawati, Ruth; Djohan, Apridah Cameliawati
BERITA BIOLOGI Vol 12, No 3 (2013)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v12i3.642

Bacterial cellulose has long been manufactured and used for industrial purposes and health. Bacterial cellulose more profitable than the cellulose plants because during the manufacturing process, they do not harm the environment. The purpose of this study was to identify differences of cellulose produced by Acetobacter xylinum and Acetobacter sp. RMG-2 from that produced by plants. The study was also aimed to determine superiority of carboxymethyl cellulose (CMC) produced by those bacteria. The medium HB was prepared for the production of cellulose from both bacteria. Bacterial cellulose preparation was carried out to obtain solid fine powder, followed by manufacturing carboxymethyl cellulose through several stages to obtain CMC powder. CMC analysis was performed for both bacteria and plants targeted on the surface structure of cellulose, the density of solids, viscosity CMC and functional groups. As a result, the surface fiber cellulose plants had a wider space than fiber cellulose bacterium. The density of solids of CMC A. xylinum, A. sp. RMG-2 and plant were 30.9998 g/cm3, 0.0079 g/cm3 and 0.9978 g/cm3 respectively. Viscosity of the CMC were of 5.78 cP, 5.25 cP and 5.91 cP for each A. xylinum,A. sp. RMG-2 and plant. CMC functional groups of bacteria has met the parameters of success as indicated by the infrared spectrum since it formed a methyl group, carboxyl group and the group of sugar. Cellulose Acetobacter sp. RMG-2 and A. xylinum cellulose can replace plants through the process of compound alkalization with sodium hydroxide, because the compound can lower the level of density of pores of cellulose fibers. The CMC resulting from bacterial cellulose as good as CMC plant and had characteristics resembling CMC plant.

MIKROBA ENDOFIT DARI TANAMAN SRIKAYA (Annona squamosa L.) SEBAGAI PENGHASIL ANTIMIKROBA Staphylococcus aureus DAN Candida albicans [Antimicrobial activity of endophytic microbes from sugar-apple (Annona squamosa l.) plant againts Staphylococcus aureus and Candida albicans] Melliawati, Ruth; Sunifah, Sunifah
BERITA BIOLOGI Vol 16, No 1 (2017)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Various studies indicated that endophytic microbes lived in the plant tissues and produced antimicrobial compounds. Sugar-apple plant Â (Annona squamosa L) contained alkaloids, cyanogenic glycosides, and flavonoids. The purpose of this reasearch were (1) to determine the endophytic microbes isolated from sugar-apple plant (2) to study inhibiting capabillity of endophytic isolate against Staphylococcus aureus and Candida albicans, (3) to analyze antimicrobial compounds produced by the potential endophytic isolate. Diffusion agar plate methode was used to assessed antimicrobial activity. Antimicrobial compounds were analyzed by Thin Layer Chormatography (TLC) and High Performance Liquid Chormatography (HPLC), compared with erythromycin, metronidazole and tetracycline. Twelve bacterial isolates and 24 fungus were isolated. Selected bacteria, BMC 1.1, showed the biggest clear zone on C. albicans culture on agar medium, meanwhile selected fungi, BTCK 1.1T, formed the biggest colony on S. aureus culture on agar medium. TLC and HPLC analysis showed that the Rf value of BMC 1.1 and BTCK 1.1T chloroform phase fractions was similiar to metronidazole. Metronidazole concentration in C1, C2, Ck1 and Ck2 fraction were 170.98 ppm, 18.27 ppm, 1.51 ppm and 4.14 ppm respectively.

Glucoamylase Production by Aspergillus awamori KT-11 In Solid State Fermentation Using Cassava Peel as Substrate Perwitasari, Urip; Nuryati, Nuryati; Melliawati, Ruth; Yopi, Yopi
ANNALES BOGORIENSES Vol 21, No 1 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

In order to utilization of cassava peel waste this study tries to produce glucoamylase by solid state fermentation with Aspergillus awamori KT-11. Composition medium and drying technique are affecting the glucoamylase production. The highest glucoamylase activities were from cassava peel plus mineral medium. Activity glucomaylase in cassava peel plus mineral medium by oven drying was 365 U/mL and freeze dring was 452 U/mL.Â It is conclud cassava peel plus mineral is a better substrate for glucoamylase production from A. awamori KT-11 in solid state fermentation. Powder of glucoamylase also proved capable of hydrolyzing starch-based biomass.Â

Isolation of Endophytic Microbes from Gunung Halimun National Park, West Java, Indonesia and Bioassay Their Potency for Eradicating Microbial Crops Pathogen Sukara, Endang; Melliawati, Ruth
ANNALES BOGORIENSES Vol 17, No 1 (2013): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/73

Gunung Halimun National Park (GHNP) â West Java, Indonesia is the largest preserved primary forest in West Java. Diversity of plants and animals of this park have been studied intensively during the 15 years years. Diversity of endophytic microorganisms, however, has never been reported. Endophytic microbes are those who reside in the interspatial tissues of plants, have a capacity to produce array of secondary metabolites. This paper illustrates the occurrence of endophyte microbes in diverse flowering plants of GHNP. Total of 160 bacteria and 337 fungus were isolated from 86 plants species in the area. Out of that, 159 bacterial isolates and 320 fungus isolates survived in our collection and tested against four major microbial crops pathogen namely Xanthomonas campestris, Pseudomonas solanacearum, Colletotrichum glocosporoides and Fusarium oxysporum cubense. Plate Agar Test Assay method reveals that 51 among 159 bacterial isolates and 62 among 320 endophytic fungal isolates have an ability to inhibit the growth of microbial crop pathogens. Endophytic bacteria can inhibit more microbial crops pathogen but the inhibition ability is less compare to that of endophytic fungi. From our preliminary study clearly shows that endophytic microbes of GHNP should have potential value in developing biological control agent to combat microbial crop pathogens and eventually reduce the use of synthetic chemicals.

PENAPISAN BAKTERI ENDOFIT PENDEGRADASI PATI DARI TAMAN NASIONAL GUNUNG HALIMUN (TNGH) Melliawati, Ruth; Sulistiyowati, Eki
Teknologi Indonesia Vol 35, No 1 (2012)
Publisher : LIPI Press

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jti.v35i1.178

In order to produce a good quality biological product such as starch, an enzyme namely amylase was crucially required. Potential microbe could generate sugar through starch degradation. Endophytic bacterias were found on various tissues of plant. Indonesia is really potential to become a source of advantagous endophytic bacteria with its rain forests. The purposes of this research were (1) to screen which endophytic bacteria could degradate starch, (2) to determine the type of reducing sugar generated and (3) to determine the value of amylase activity and theaccumulation of reducing sugar. This research was expected to give some information about the use of endophytic bacteria as an alternative of sugar producer for several industrial sectors. The sceening was practiced to the 30 isolated endophyt bacteria from the plants in Halimun Mountain National Park. Those bacteria was inoculated at a medium named Potato Starch Agar (PSA). The ability to degradate starch could be known when there were clear zones around them. The kinds of sugar were detected by TLC method while the analysis of the amount of reducing sugar and experiment about the activity of amylase were done based on Somogy-Nelson method. The sceening test result that 12 isolates bacteria could grow well on PSA. Three of them generated large clear zones. They wereHL.72 B.153, HL.74 B.156 and HL.89 B.197, with the width of each clear zone were 5,31 cm2, 2,69 cm2 and 6,6 cm2. The experiment of enzymatic activity and the accumulation of of reducing sugar of three choosen isolated bacterias showed that the highest result was HL.89 B.197 with 556,67 Unit and its sugar accumulation value was 631,89 ppm. Then the amylase activitys value of HL.72 B.153 was 505,3 Unit and its sugar accumulation value was 472,29 ppm. The last result was HL.74 B.156 with the amylase activitys value was 263,73 Unit and its sugar accumulation value was 330,21 ppm. The result of TLC showed that a fermentation of HL.72 B.153 was Rf=0,42 of Rf value was same as galactose. HL.74 B.156 Rf value was 0,54 was same as the Rf value of fructose and the Rf value of HL.89 B.197 was 0,54 which was same as the Rf value of glucose.

Enzyme Production From Cassava Peels by Aspergillus Awamori KT-11: The Making of Natural Sweetener From Several Tubbers Melliawati, Ruth; Rahman, Farida
ANNALES BOGORIENSES Vol 23, No 1 (2019): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

The use of cassava (Manihot esculenta Crantz) peel for enzyme production has not been widely used. The purpose of this study was to produce complex amylase enzymes from cassava peel by A. awamori KT-11 and apply them in the manufacture of natural sweeteners. Enzyme production is carried out on red and white cassava peel. Media of cassava peel sterilized, inoculated with 1% A. awamori KT-11, incubated for 5 days, then dried at 50°C and mashed. Making sugar is done on cassava flour, sweet potato ( Ipomoea batatas L), taro (Colocasia esculenta) and cocoyam (Xanthosoma sagittifolium) with different concentrations of 10%, 15%, 20%, and 15% and 20% enzyme concentrations. The hydrolysis process is carried out for 3 days at 60°C. The enzyme activity in red cassava peel was 405,006 U/mL and white cassava peel was 321,239 U/ml. The sugar produced in cassava, taro, sweet potato, and Cocoyam was 101.38 mg/mL, 81.18 mg/mL, 55.929 mg/mL, and 42.874 mg/mL, respectively. The results of TLC showed that cassava and taro sugar contain maltose, lactose and glucose, sweet potatoes contained glucose and dextrin and Cocoyam containing fructose. The sweetness level of sugar from cassava, taro, sweet potato and Cocoyam is 14 brix, 12 brix, 9 brix and 9 brix, respectively.

Senyawa Antibakteri Escherichia coli ATCC 35218 dan Staphylococcus aureus ATCC 25923 dari Kapang Endofit Taman Nasional Gunung Halimun Melliawati, Ruth; Harni, Harni
Jurnal Natur Indonesia Vol 12, No 1 (2009)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Endophytic fungus is a microorganism which live in the interstitial spaces healthy tissues of the host plant, andhas capability to produce secondary metabolite such as micotoxin, antibiotic and antiviral. This research wasaimed to find out the isolates of endophytic fungus produce antibacterial compounds to inhibit Escerichia coliATCC 35218 and Staphylococcus aureus ATCC 25923, and to investigate the Retention time (Rt) of the antibacterialcompounds produced by endophytic fungus with High Performance Liquid Chromatography (HPLC) methods.Diffusion Agar Plate Method was used to examine the antibacterial compounds on Escerichia coli ATCC 35218 andStaphylococcus aureus ATCC 25923. While the antibacterial compounds were examined with Thin LayerChromatography (TLC) and High Performance Liquid Chromatography (HPLC) methods, and the result werecompared with Chloramphenicol and Ampicillin antibiotic standart. Two isolates of endophytic fungus namelyHl.46F.211 and HL.57F.258 were inhibited the growth Escherichia coli ATCC 35218 and three isolates namelyHL.48F217, HL.53F.243 and HL.57F.258 showed antagonistic action against Staphylococcus aureus ATCC 25923.The results of TLC and HPLC analysing method show that antibacterial compounds produced by endophytic fungusHL.46F.211 had Rt (Retention Time) rate similar with antibiotic Chloramphenicol at 2,796 (at water fraction) and Rtantibiotic Amphycillin at 2,662 (at Chloroform fraction). While antibacterial compounds produced by endophyticfungus HL.57F.258 had Rt rate similar with antibiotic Amphycillin at 2,650 (at Chloroform fraction).

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