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ISOLATION AND IDENTIFICATION OF MICROORGANISMS DURING SPONTANEOUS FERMENTATION OF MAIZE [Isolasi dan Identifikasi Mikroorganisme pada Fermentasi Spontan Jagung] Rahmawati, .; -Hariyadi, Ratih Dewanti; Hariyadi, Purwiyatno; Fardiaz, Dedi; Richana, Nur
Jurnal Teknologi Dan Industri Pangan Vol 24, No 1 (2013): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.6066/6953

Abstract

ISOLATION AND IDENTIFICATION OF MICROORGANISMS DURING SPONTANEOUS FERMENTATION OF MAIZE [Isolasi dan Identifikasi Mikroorganisme pada Fermentasi Spontan Jagung] Rahmawati1,2), Ratih Dewanti-Hariyadi1,3)*, Purwiyatno Hariyadi1,3), Dedi Fardiaz1,3) and Nur Richana4) 1) Department of Food Science and Technology, Bogor Agricultural University, Kampus IPB Darmaga, Bogor, Indonesia 2) Department of Food Technology, Sahid University, Jakarta, Indonesia 3) Southeast Asia Food Agricultural Science and Technology (SEAFAST) Center, Bogor Agricultural University, Kampus IPB Darmaga, Bogor, Indonesia 4) Indonesian Center for Agricultural Post Harvest Research & Development (ICAPRD), Bogor, Indonesia   Accepted April 05th 2013 / Approved June 12th  2013 ABSTRACT   Maize was traditionally the second most common staple food in Indonesia. Conversion to maize flour has been accomplished to improve its convenience. Traditionally, maize flour is produced by soaking the kernels in water followed by grinding. It was reported that final physicochemical characteristics of the maize flour were influenced by spontaneous fermentation which occurred during soaking. This research aimed to isolate and identify important microorganisms that grew during fermentation thus a standardized starter culture can be developed for a more controlled fermentation process. Soaking of maize grits was conducted in sterile water (grits:water=1:2, w/v) in a closed container at room temperature (±28ºC) for 72 hours. After 0, 4, 12, 24, 36, 48, 72 hours, water and maize grits were sampled and tested for the presence of mold, yeast, and lactic acid bacteria (LAB). Isolates obtained from the spontaneous fermentation were reinoculated into the appropriate media containing starch to observe their amylolytic activity. Individual isolate was then identified; mold by slide culture method, while yeast and LAB by biochemical rapid kits, i.e. API 20C AUX and API CH50, respectively. The number of each microorganism was plotted against time to obtain the growth curve of the microorganisms during spontaneous fermentation. The microorganisms were identified as Penicillium chrysogenum, P. citrinum, A. flavus, A. niger, Rhizopus stolonifer, R.oryzae, Fusarium oxysporum, Acremonium strictum, Candida famata, Kodamaea ohmeri, Candida krusei/incospicua, Lactobacillus plantarum 1a, Pediococcus pentosaceus, L. brevis 1, L. plantarum 1b, and L. paracasei ssp paracasei 3. Four molds and one yeast were amylolytic while none of the LAB was capable of starch hydrolysis. The growth curve suggested that the amylolitic mold and yeast grew to hydrolyze starch during the course of fermentation, while the LABs benefited from the hydrolyzed products and dominated the later stage of the fermentation.  
Purification of alfa-Amylase from Bacillus stearothermophilus by Ultrafiltration Membrane Lestari, Puji; Richana, Nur; Murdiyatmo, Untung
Jurnal Mikrobiologi Indonesia Vol 5, No 1 (2000): JURNAL MIKROBIOLOGI INDONESIA
Publisher : Jurnal Mikrobiologi Indonesia

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The objective of this experiment was to evaluate the optimum conditions of cc-amylase of Bacillus slearotherinophiluspurification by ultrafiltration and the step of a-amylase purification from indigenous isolate. Culture filtrate frombioreactor was separated by microfiltration membrane with a pore sIze of 0.2 tm. The aupernatant was purifie again byultrafiltration system membrane with cut off 30 000 Dalton. Treatments of cz-amylase purified by ultrafiltration werecarried out at flow rate of 30,45 and 60 mI/minute, and concentrated by about 5, 10 and 15 tImes. The crude enzymesresulted From ultrafiltration were precipitated with acetone. The results showed that the optimum condition ofultrafiltration was using flow rate of 30 mI/minute and concentrated by about 10 times. At the optimum condition ofultrafiltration, the specific activity of ci-amylase was of 6 686.6 U/mg with 2.3 fold purification factor. The effect of flow ratedecreased the total enzyme activity, specific activity and yield. The concentration disposal could decrease total activity andprotein, but not always reduced specific activity of the enzyme. Purification of crude enzyme by ultrafiltration and acetonereduced the total activity, total protein and yield, but specific activity and purification factor increased. Ultrafiltrationfollowed by acetone precipitation, gave enzyme specific activity of 18155.4 U/mg, purification factor of 6.3 and yield of20%, respectively. Zymogram analysis using Native-Polyacrylamide Agarose Gel Electrophoresis indicated´ ci-amylase ofapproximately 192 932.8 Da.
Study of the Addition Calcium Ion to alfa-Amylase Activity and Stability from Bacillus stearothermophilus TII 12 ., Rosminik; Richana, Nur; Lestari, Puji; Damardjati, Djoko Said
Jurnal Mikrobiologi Indonesia Vol 6, No 1 (2001): JURNAL MIKROBIOLOGI INDONESIA
Publisher : Jurnal Mikrobiologi Indonesia

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The effcet of storage in liguld or freeze dried condition and calcium ion addition to activity md stability of asmylase from Bacillus stewotkcrmophilhis TlI. have been carried out. The enzyme was stored under several condition:I enzyme was freeze dried, ii enzyme was precipitated by aceton and resoluble in phosphate citrate buffer, iiienzyme was freeze dried after aceton precipitation and reaolibilizstioii in phosphate citrate buffer, Iv enzymeprecipitated by *ceton, resoluble in phosphate Citrate buffer and added by 5 mM calcium ion, and v enzyme wasfreeze dried after aceton precipitation, resolibilization in phosphate citrate buffer and added by 5 mM calcium ion.ReuIts show after nine month storage that the beat enzyme obtained in the addition of calcium ion in the freezedried condition 79.5%.
Cultivation of Thermophilic Bacteria Isolate of alfa-Amylase Production Richana, Nur; Yusuf, Gagan Maulana; Lestari, Puji; Damardjati, Djoko Said
Jurnal Mikrobiologi Indonesia Vol 4, No 2 (1999): JURNAL MIKROBIOLOGI INDONESIA
Publisher : Jurnal Mikrobiologi Indonesia

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Cultivation of thermophilic bacteria isolate TVII-6 for a.amylase production. Thermophilic bacteria TVIJ-6 was isolatedand selected from the soil sample taken from Dieng vulcanic. The specific activity of ainylase produced by TVfl-6 was 1624.37U/mg protein. Cassava starch was used as carbon source on enrichment culture and the optimum concentration was 1% withspecific activity 2092.23 U/mg protein. Cultivation in a 2-I bioreactor at pH 7.0 and temperature of 50°C, combine with 250 rpmagitation and 1.0 vvm aeration produced the highest a.amylase. The maximum specific growth rate obtained was 0.147/hours.The relationship between product and substrate was linear Y=0.129 X + 0.024. Efficiency of the substrate to produce amylaseY, was 0.129 g proteinlg substrate. Relationship between the bacterial growth and substrate was Y = 03115 X + 0.013.Efficiency of the substrate for growing the bacteria Y,, was 0.315 g cell/g substrate. Based on the bacterial growth and amylaseproduction, amylase could be considered growth associated, with kinetic parameter of product ratep was 0.37gfgfhr.
Hydrolysis Methods of Starch for Culture Media of Bacillus sp. BMN14 Producing Biosurfactant Lipopeptide Richana, Nur; Suryani, Ani; Makagiansar, Helena Yusuf; Irawadi, Tun Tedja
Jurnal Mikrobiologi Indonesia Vol 5, No 2 (2000): JURNAL MIKROBIOLOGI INDONESIA
Publisher : Jurnal Mikrobiologi Indonesia

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Ezymatic and acid hydrolysis of starch for culture media of Bacillus sp. BMN14 producing lipopeptide blosurfactantwere studied using cassava, arrowroot and sago starch. These starch bydrolysates were used as glukose substitute Inthe were to culture media of biosurfactant.producing bacteria. Starch hydrolyzed from arrowroot 73.08-83.5% andcassava 80.0-87.98% yielded glucose higher than sago 62.5-64.5%. Capability of the bacteria to produce biosurfactantrepresented as the amount of acid precipitate 0.78 g/l and surface tension of the broth 33.6 mN/rn in the culturemedium contained enzymatic hydrolyzed-starch.
The Selection of Xylanase-Producing Indegenous Bacteria Richana, Nur; Lestari, Puji; Thontowi, Ahmad; ., Rosmimik
Jurnal Mikrobiologi Indonesia Vol 5, No 2 (2000): JURNAL MIKROBIOLOGI INDONESIA
Publisher : Jurnal Mikrobiologi Indonesia

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The isolation and selection of xylanase producing bacteria have been done from soil and waste of starch Industry.Strains were cultivated in neutral and alkaline media with xilan as a substrate. Colonies which produced clearingzone were presumed as xylanolytic bacteria and chosen for further screening. Results show that 10 isolates ofbacteria In neutral condition and 25 bacteria in alkaline condition. There are three isolates in neutral condition ON.12, ON-13 and ON-33 have high enzyme activities at 19.93, 23.42 and 19.32 U/ml, and specific activities which were80.37, 71.85 and 114.36 U/mg protein respectively. On the other hand there are two isolates in alkaline condition TAIS and TA-fl15 which have high enzyme activities at 12.43 and 11.18 U/mI, and specific activities which were 36.72 and42.68 U/mg protein respectively.
Analysis of Reducing Sugars on Hydrolysis of Cassava Starch by a Thermostable a-Amylasefrom Bacillus stearothemophilus TII 12 Lestari, Puji; Darwis, Abdul Aziz; Syamsu, Khaswar; Richana, Nur; Damardjati, Djoko Said
Jurnal Mikrobiologi Indonesia Vol 6, No 1 (2001): JURNAL MIKROBIOLOGI INDONESIA
Publisher : Jurnal Mikrobiologi Indonesia

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The product of a-amylase hydrolysis was evaluated for their application In sugar syrup Industry. The objectives ofthis experiment were to determine the enzyme production of Bacillus stearothermophilus TH,2 in a five liter bioreactorand to analyze its hydrolysis product on cassava starch using thin layer chromatography and high performance liquidchromatography. The bacterium was cultured in the bloreactor for 48 hours, and then the biomass, enzyme activity,protein and reducing sugar contents in the filtrate were evaluated In the course of cultivation. The strain secreted anextraceiiuiar a-amylase in the optimal condition at pH 6.5, 5O´C, agitation of 300 rpm and aeration of 1.5 vvm for 24hours. The highest activity of a.amylase and reducing sugar content of 1 068.87 U/mI and 4.48 g/l respectively wereobtained after 24 hours incubation. Hydrolysis products by the crude enzyme on cassava starch were evaluated atdifferent Incubation time. In the course of incubation the content of glucose, dextrin, maltose and oligosaccharideswere increasing. After 24 hours the concentration of glucose and maltose reached 51 970 and 10 090 ppm respectively.Based on the enzymatic products, we concluded that thermostabie a.amylsse produced by B. slearothermophilus was aaendo-a-amylase.
The Process of Xylanase Production from Bacillus pumilus RXAIII-5 RICHANA, NUR; IRAWADI, TUN TEDJA; NUR, M. ANWAR; SAILAH, ILLAH; SYAMSU, KHASWAR
Microbiology Indonesia Vol 1, No 2 (2007): August 2007
Publisher : Indonesian Society for microbiology

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The optimum conditions for the growth of Bacillus pumilus RXAIII-5 (a potential xylanase producer) were sought, these included temperature, pH, aeration, and agitation of the culture batch. Afterwards a mathematical model based on the parameter of cultivation kinetics was formulated. At the same time, the rheology of the fluid used for bacterial cultivation in a bioreactor was studied. The data obtained was used for estimating the ‘scaling up’ of enzyme production. The results of the study indicate that the optimum condition for processing in 50 ml Erlenmeyer flask are used temperature of 35 oC (308oK), pH 7, and an agitation rate of 140 rpm. The highest xylanase activity and its specific activity are 297.132 U.ml-1 and 655.32 U.g-1protein, respectively. Subsequent experiments in a bioreactor using all of the experiment parameters mentioned above, except for the agitation rate, shows that the results are as follows. The highest specific growth was at 0.082 hour-1 at an aeration and agitation rate of 0.5 vvm and 150 rpm, respectively. Based on the data of the cultivation kinetics, the optimum conditions for the fermentation in Biostat 2L-bioreactor is 1 vvm and 200 rpm of aeration and agitation, respectively . The efficiency of substrate (Yp/s) and of cell biomass (Yp/x) to produce xylanase is 50.744 U.g-1 and 43.906 U.g-1, respectively. The efficiency of substrate to cell production (Yx/s) is 1.178g.g-1. The liquid cultivation-medium has non-Newtonian properties. Based on a mathematical model it is found that the consistency index (k constant) and index of liquid behavior (n value) are 0.179 g.cm-1.second-1 and 0.3212, respectively. Becouse the value of 0
POTENTIAL USE OF AN EXTRACELLULAR ENZYME OF a-AMYLASE FROM INDIGENOUS INDONESIAN MESOPHILIC BACTERIA Lestari, Puji; Richana, Nur; Masriani, Rina
Indonesian Journal of Agricultural Science Vol 14, No 1 (2013): April 2013
Publisher : Indonesian Agency for Agricultural Research and Development - MOA

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Amylase enzyme has a great significance for industrial usages in  Indonesia. However, this enzyme is still imported. The use of bacteria in biotechnological process of industrial products such as enzyme production has stimulated the exploration of extracellular amylase producing  bacteria. This study aimed to identify and analyze the potential use of amylolytic bacterial enzymes for hydrolyzing cassava starch. Two bacterial isolates, i.e. MII-10 and DKW-8 originated from Indonesia soil were identified based on their morphological, physiological and biochemical properties according to the standard protocol. The isolates were then  cultivated on fermentation medium and their growth pattern and  enzymatic assays were observed. The acetone-precipitated crude enzyme harvested based on predetermined cultivation time was used for  enzymatic hydrolysis product characterization on cassava starch using thin layer chromatography (TLC). The results showed that the mesophilicbacteria isolates (MII-10 and DKW-8) were belonged to Bacillus licheniformis. The maximum bacterial cell growth and enzyme activity were reached at 48 hours after incubation. The MII-10 isolate was found more stable than DKW-8 in producing amylase enzyme. Amylase produced by the MII-10 and DKW- 8 isolates was identified to be an endo-a-amylase as confirmed by oligosaccharides and dextrin of the random hydrolysisproducts. Relatively high dextrose equivalence (DE) value of a-amylase of MII-10 (DE of 9.96) suggests that the enzyme is prospective for  saccharification of starchy material in glucose syrup industry.
Biosorpsi ion tembaga dalam limbah tailing menggunakan jamur pelapuk putih amobil Biosorption of copper ion in tailing mining effluent using immobilizede white-rot fungi DIMAWARNITA, Firda; SUHARYANTO, .; TRI-PANJI1, .; RICHANA, Nur; ZAINUDIN, Achmad
E-Journal Menara Perkebunan Vol 83, No 1: Juni 2015
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v83i1.11

Abstract

Abstract         Copper and gold mining industry produce large amount of tailings effluent containing heavy metals such as Cu and Hg that will polute  environment and agricultural land, if it is not properly treated.  Effort in reducing pollution level and recovery of metal in tailing could be conducted with biosorption process using microbial cell as an absorbent agent. This study aims to determine the ability of selected white rot fungi (WRF) immobilized on empty fruit bunches of oil palm (EFB)  to absorb Cu (II) metals in tailing. Based on growth rate, ligninolytic enzymes activity, and the absorption capacity of heavy metals, the superior WRF candidate was Omphalina sp. The ability of biomass Omphalina sp. to decrease the con-centrateion of Cu (II) with an initial concentration of 150 ppm, up to 63%. Omphalina sp. that immobilized on TKKS was able to reduce Cu (II) as much as 66-77% at pH 4.0 for 60 minutes. Abstrak                Eksploitasi tambang tembaga dan emas banyak menghasilkan limbah tailing yang masih me-ngandung logam berat seperti Cu (II) dan Hg (II). Limbah tersebut berpotensi mencemari perairan dan lahan pertanian bila tidak ditangani dengan baik.  Usaha untuk mengatasi limbah tailing dan sekaligus memekatkan (recovery) logam di dalam-nya dapat dilakukan dengan proses biosorpsi menggunakan sel mikroba. Penelitian ini bertujuan menetapkan kemampuan biomassa jamur pelapuk putih (JPP) yang diimobilisasi pada tandan kosong kelapa sawit (TKKS) dalam mengabsorpsi ion logam Cu (II). Metode biosorpsi logam yang digunakan dengan sistem batch dengan kadar logam 300 ppm, 200 ppm,dan 150 ppm. Hasil seleksi JPP berdasarkan laju pertumbuhan, aktivitas enzim ligninolitik, dan penyerapan logam berat telah  diperoleh   kandidat   JPP   unggul  yaitu Omphalina sp. Dalam media PDB,  Omphalina sp. toleran terhadap Cu (II) hingga konsentrasi 150 ppm. Kemampuan biomassa Omphalina sp. untuk penurunan konsentrasi Cu (II) dengan konsentrasi awal 150 ppm yaitu mencapai 63%. Omphalina sp. yang diimobilisasi dengan TKKS mampu menurun- kan kandungan Cu (II) sebesar 66-77%  pada pH 4.0 selama 60 menit.