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"Inoculum for making tempe is usually made from the mould Rhizopus oryzae or Rhizopus oligospons. Large scale use of an inoculum from one source only (LKN-LIPI) might threaten the biodiversity of microorganisms in traditionally made inoculum. The potential use of other Rhizopus moulds as inoculum for tempe production should be examined. Rh. microsporus var. rhizopodifonnis UICC 520 isolated from Hibiscus from Manado and Rh. microsporus var. chinensis UICC 521 isolated from tempe from Aceh were examined for their potentials to produce tempe.Rhizopus has also been known to be able to transform antioxidants such as isoflavone in tempe. The potential of Rhizopus microsporus v. Tiegh. var. chinensis (Saito) UICC 521 to transform glucoside isoflavones in soybean tempe has been evaluated.The results showed that Rh. microsporus var. rhizopodiformis UICC 520 has little potential as a tempe inoculum. Rh. microsporus var. chinensis UICC 521, however, is very potential as an inoculum to produce good tempe and produces abundant spores ((3-10)x106 cell/ml) on rice substrate.Two types of aglycon isoflavone, daidzein and genistein, were detected by TLC separation. The total isoflavone concentration during fermentation period of 24, 48, 72 hours was determined using HPLC. At 24 hours the concentration was 0.142 mg/g tempe, while at 48 and 72 hours the concentration were 0.181 and 0.135 mg/g tempe, respectively. The highest total isoflavone concentration was detected at 48 hours fermentation period. Hence, there was an increase of aglycon isoflavones up to 6000% after transformation of isoflavones in soybean by Rh. microsporus var. chinensis UICC 521 when compared to the initial concentration before transformation."

"SUMMARY This research was carried out to study the diversity of Rhizopus in Irian Jaya. Tempe samples were collected from regions in Irian Jaya. The aim of this research was to Isolate and Identify Rhizopus spp. from traditionally made tempe, to make usar from the Rhizopus sp. isolates, and to produce tempe from the chosen usar. Isolation was done using direct plating method. Nineteen Rhizopus spp. isolates have been isolated from soybean tempe collected from several regions in Irian Jaya. Examination of their morphological characteristics, growth temperature, and spore ornamentation, showed that eighteen isolates belong to the Rh. microsporus group: two isolates were Rh. microsporus var. oligosporus, sixteen isolates were Rh. microsporus var. chinensis. One isolate was Rh. oryzae. The isolates were examined for their ability to produce usar. Out of the nineteen isolates that were tested, only seven produced good usar (inoculum): six isolates from Rh. microsporus var. chinensis, and one of Rh. oryzae. The amount of spores before and after inoculum production were determined by Total Plate Count (TPC) method. The average of viable spores from all isolates before the inoculum production was ((2.5-3.2)x14' cetllmly, and after the inoculum production was ((1.2-1.8)x104 celllg). The usars (Inocula) from seven Isolates were examined to produce tempe. The Rh. oryzae inoculum produced better tempe than the Rh. microsporus var. chinensis Inoculum based on colour, texture, aroma and shelf-life."

"Yotefa Bay, lying on the north coast of Jayapura, was established as a Tourist Park in 1978, based on the Minister of Agriculture's Decree No.: 372/KptslUm/6/1978 dated 9"? June 1978, Mangrove forest, which spreads out from the north part to southeast part of Yotefa Bay, is dominated by Rhizophora spp. Research on pattern of litter production of Rhizophora mucronata in Yotefa Bay Tourist Park, Jayapura-Papua, was conducted from June - September 1999. This research was experimental research using vegetation analysis transect method, and the data analysis to know the community srtucture was conducted using quantitative parameters, i.e.: diameter at breast high (dbh); density and bassal area. Litterfall was collected according to the methods describes by Brown (1984), using a spesilic litter traps- All materials acumulated in the traps were collected once per week during the sampling period. Estimated of litterfall rates were using the formulas by Chansang and Poovachiranon (1990), and estimated of production of litterfall using the statistical porcedures by Walpole (1985).

The results showed that total of R. mucronata litter production ranged between 18.8120 tons ha"year" to 78.84 tons ha"year". The largest component of litter obtained was leaves: 52.04%, followed by component of fruits 34.34%, flowers 9.91% and skin-branches 3.78% respectively. There is not a tendency that pattern of leave litter production in the mangrove forest in Yotefa Bay follows rainfall fluctuation, and did not tend to be greater during rainy season. lt is necessary to conduct similar research in other mangrove forest of Yotefa Bay Tourist Park. It is hoped that in th next research, besides the use of more spesific supporting parameters, the location for observation will be wider, with many more litter traps and longer time for observation. Research on decomposition rate of R. mucronata leave litter was conducted in 16 weeks, starting from June to September 1999.

This research was also a factorial experiment with two factors. Data was analyzed using ANOVA with two-way classification in order to know whether the time-length of soaking and immersion, and the location of decomposition have any influence on the rate of decomposition. Analysis was also done to determine the rate of decomposition value (k value). Chemical analysis was done in the laboratory to know the content of Carbon, Nitrogen, Phosphor and Potassium elements in the leave litter of the R. rnucronafa of the decomposition remnants.

The results showed that decomposition rate of leave litter of R. mucronata was influenced by the time-length of soaking and immersion and the decomposition location. The rate of decomposition value (k value) average was 1.19; this value showed that decomposition worked well. Average contents of element of Carbon, Nitrogen, Phosphor and Potassium were respectively 14,135, 1,27%, O,100% and O,212%. The existence of those elements in the decomposed mangrove leave litter has effected the rate of decomposition value. lt is necessary to conduct a similar research in the other part of mangrove forest in the Yotefa Bay Tourist Park, Jayapura. lt is expected that the next research will be focused more on microorganism activities (fungi and bacteria) and the influence of other abiotic factors, which have not been done in this research, in order to know the influence of those factors onthe rate of decomposition."

"ABSTRAKProtease asam yang dihasilkan oleh kapang memiliki peranan penting dalam industri pangan dan farmasi. Rhizopus spp. merupakan salah satu jenis kapang yang memiliki aktivitas proteolitik dan tidak menghasilkan toksin. Kemampuan Rhizopus dalam menghasilkan enzim proteolitik bervariasi baik antar spesies maupun antar galur dalam spesies yang sama. Untuk memperoleh enzim dengan aktivitas proteolitik yang tinggi, perlu diperhatikan faktor lingkungan yang mempengaruhi aktivitasnya. Tingkat keasaman (pH) dan suhu merupakan faktor lingkungan yang sangat menentukan aktivitas enzim proteolitik. Setiap enzim memiliki pH optimum yang khas, yaitu pH lingkungan yang menyebabkan aktivitasnya maksimum. Enzim yang dihasilkan oleh mikroorganisme juga mempunyai suhu optimum tertentu untuk mengkatalisis suatu reaksi enzimatis. Indonesia dikenal kaya akan keanekaragaman hayati kapang. Untuk memanfaatkan keanekaragaman hayati kapang indigenous Indonesia, perlu dilakukan penelitian untuk mengetahui potensi produksi protease asam ekstra selular dari Rh. microsporus v. Tiegh. var. rhizopodiformis (Cohn) Schipper & Stalpers dan Rh. microsporus v. Tiegh. var. chinensis (Saito) Schipper & Stalpers koleksi University of Indonesia Culture Collection (UICC). Penelitian ini bertujuan mengetahui potensi biakan Rh. microsporus var. rhizopodiformis UICC 519, 520, dan Rh. microsporus var. chinensis UICC 521 dalam menghasilkan enzim proteolitik, dan penentuan waktu fermentasi, pH, dan suhu optimum aktivitas proteolitik pada kapang terpilih. Aktivitas proteolitik semi kuantitatif dari ketiga biakan dipelajari berdasarkan kemampuannya membentuk zona bening pada medium 4% (b/v) Skim Milk Agar (SMA) dengan cara pencawanan (plating) spora tunggal pada suhu ruang selama 28-29 jam. Filtrat kultur fermentasi Rh. microsporus var. rhizopodiformis UICC 520 yang diekstraksi dari medium dedak padi digunakan untuk uji aktivitas proteolitik. Uji aktivitas proteolitik kuantitatif dari filtrat kultur dilakukan dengan mengukur jumlah tirosin yang dihasilkan dari hidrolisis substrat kasein pada suhu suhu 37°C ± 2°C selama 30 menit. Setiap labu yang berisi medium fermentasi steril diinokulasi dengan 1 ml suspensi spora Rh. microsporus var. rhizopodiformis UICC 520 yang berisi 3,9-4,6 x(10 pangkat 5)koloni/ml. Medium fermentasi selanjutnya diinkubasi pada suhu ruang (26--30°C) tanpa pengocokan. Protease kasar diekstraksi dari medium fermentasi pada 0, 24, 48, 72, 96, dan 120 jam untuk mengetahui waktu fermentasi optimum. Untuk menentukan pH optimum aktivitas proteolitik, kasein sebagai substrat enzim dilarutkan dalam larutan dapar hingga diperoleh pH 2,0; 3,0; 4,0; 5,0; 6,0; 7,0; dan 8,0. Untuk menentukan suhu optimum aktivitas proteolitik, sampel direaksikan dengan substrat kasein 0,7% (b/v) dalam dapar glisin-HCI pH 3,0 dan dinkubasikan pada suhu 35, 40, 45, 50, dan 55°C. Berdasarkan diameter zona bening yang terbentuk dari pertumbuhan spora tunggal pada medium SMA dapat disimpulkan bahwa Rh. microsporus var. rhizopodiformis UICC 520 memiliki kemampuan menghasilkan protease lebih cepat dibandingkan Rh. microsporus var. rhizopodiformis UICC 519 dan Rh. microsporus var. chinensis UICC 521 dalam waktu yang sama (28-29 jam). Aktivitas proteolitik Rh. microsporus var. rhizopodiformis UICC 520 dengan medium fermentasi dedak padi mencapai maksimum, yaitu sebesar 0,0944 U/ml pada inkubasi selama 72 jam. Peningkatan aktivitas proteolitik yang cepat diikuti dengan penurunan pH filtrat kultur. Enzim proteolitik dalam filtrat kultur Rh. microsporus var. rhizopodiformis UICC 520 aktif pada kisaran pH substrat 2,0-6,0 dan memiliki dua puncak aktivitas, yaitu puncak tertinggi pada substrat kasein dengan pH 2,0-3,0 (0,0772--0,0912 U/ml) dan puncak aktivitas kedua yang lebih rendah dari puncak pertama, yaitu pada pH substrat 5,0-6,0 (0,0603-0,0649 U/ml). Enzim proteolitik dari filtrat kuitur Rh. microsporus var. rhizopodiformis UICC 520 aktif pada kisaran suhu 35--50°C. Aktivitas proteolitik tertinggi diperoleh pada suhu 45° C.

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"Thesis Supervisors: Dr. Suyanto Pawiroharsono; Prof. Dr. Indrawati GandjarSUMMARYFood deterioration is often due to lipid oxidation, excluding bacterial andenzymatic spoilage. The end-products of lipid oxidation, such as aldehydes,ketones, and alcohols are responsible for unacceptable off-tiavors and off-odors in food. Lipid oxidation can be inhibited by antioxidants.Soybean tempe is the most popular indigenous fermented food inIndonesia. Soybeans are known to contain isotiavones. Four major forms areknown respectively as acetylglycosides, malonylglycosides, glycosides, andaglycones.Tempe were produced from soybean fermentation by Rhizopus oryzaeUICC 524 and Rhizopus microsporus var. chinensis UICC 521. The tempesamples were extracted with methanol and the extraction defatted with hexa»ne. The isoflavone aglycones were isolated using column chromatography,and then anaiyzed using a gradient elution reverse phase of high-pressureliquid chromatography (HPLC). After HPLC analysis, isotiavone aglyconeswere evaporated to dryness and added to soybean oil at the 100, 200, 300,iii ' and 400-ppm concentration in test tubes, then heated at 170°C for 30 minu-tes. The oxidation of soybean oil was measured using the thiobarbituric acid(TBA) test. The result, called thiobarbituric acid reactive substances(TBARS) value, was expressed as pmolll and compared to the syntheticantioxidant, buthylated hidroxytoluene (BHT), at the same concentration.The profile of isoflavone aglycones isolated contains daidzein andgenistein. No factor-2 (6,7,4'-trihidroxyisoflavone) and glycitein were detec-ted. Daidzein resulted from biotransformation of daidzin was dominant inboth tempe samples. The isoflavone biotransformation was much greater byR. microsporus var. chinensis UICC 521 than by R. oryzae UICC 524, exceptfor the 24 hours incubation. After 72 hours of incubation, the total isoflavoneaglycones in tempe using R. microsporus var. chfnensis UICC 521 was721.6 pglg and when using R. oryzae UICC 524, 268.2 p.glg.The oxidized soybean oil without any antioxidants had a TBARS value of327.32 1 20.31 pmol/1. Addition of the antioxidants showed a decreasedTBARS value following increasing concentration for both. For concentrationuntil 300 ppm, the TBARS values of oxidized soybean oil added with isofla-vone aglycones were greater than when added with BHT, respectively55.40 zl: 2.77 pmol!! and 45.20 i 2.63 pmolll. However at concentration of400 ppm, the TBARS values of oxidized soybean oil added with isoftavoneaglycones and added with BHT did not show a significant difference.ix + -51 pp; 6 append; 5 plates; 3 tables.Bilb 35 (1964-1999).iv"

"Temulawak (Curcuma xanthorriza Roxb) memiliki kandungan utama kurkuminoid dan xanthorrhizol yang dapat memberikan manfaat kesehatan bagi tubuh. NADES merupakan green solvent yang banyak diuji coba untuk mengekstraksi senyawa pada temulawak. NADES diketahui dapat meningkatkan solubilitas dan bioavailabilitas senyawa tidak larut air seperti kurkumin. Penelitian ini bertujuan untuk mencari kombinasi pelarut terbaik, menetapkan kondisi optimum ekstraksi dengan NADES menggunakan Ultrasound-Assisted Extractionuntuk senyawa kurkuminoid total (CUR) dan xanthorrhizol (XNT) pada rimpang temulawak, dan membandingkan hasilnya dengan ekstraksi konvensionalmaserasi-etanol. Variabel yang digunakan untuk optimasiberupa persentase penambahan air pada NADES (%), waktu ekstraksi (menit), dan rasio serbuk dengan pelarut (S/L). Semua variabel dimodelkan dengan menggunakan Response Surface Methodology (RSM). Penetapan kadar dianalisis menggunakan KLT densitometri yang parameternya telah divalidasi, fase gerak diklorometana-kloroform (4:6), dan dideteksi pada panjang gelombang 425 nm (CUR) dan 224 nm (XNT). Dari hasil analisis ekstraksi, NADES terbaik yaitu kolin klorida dengan asam malat (ChCl-MA) dengan rasio 1:1. Ditemukan kondisi optimum dari hasil rekomendasi RSM, yaitu pada penambahan air 27,7%, waktu ekstraksi 17,5 menit, dan rasio serbuk pelarut 1:18,5 dengan hasil kadar 4,76 mg/g total CUR dan 12,98 mg/g XNT, sedangkan maserasi-etanol 96% menghasilkan kadar 1,88 mg/g total CUR dan 9,80 mg/g XNT. NADES-UAE lebih efektif menarik senyawa CUR dan XNT dibandingkan maserasi-etanol 96%. Data pada penelitian ini berguna untuk pengembangan metode ekstraksi hijau lebih lanjut untuk mengekstrak kurkuminoid dan xanthorrhizol menggunakan NADES.

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The Javanese turmeric (Curcuma xanthorriza Roxb) contains curcuminoids (CUR) and xanthorrhizol (XNT) as a main compounds that can provide health benefits. NADES is one of the green solvent that has been tested for extracting temulawak. This study aims to obtain the optimum condition to extract total CUR and XNT from Curcuma xanthorriza Roxb. using organic acid-based NADES with Ultrasound-Assisted Extraction. The variables used were water addition (%), extraction time (min), and solid-to-liquid ratio (S/L). All variables were modelled by using Response Surface Methodology (RSM). Determination of marker content was analysed using TLC Densitometry that was validated, dichloromethane-chloroform (4:6) as mobile phase, was detected at a wavelength of 425 nm for total CUR and 224 nm for XNT. Three organic acid-based NADESs were screened to find one NADES combination that gives the highest content of CUR and XNT. It resulted in choline chloride and malic acid DES (ChCl-MA) at a 1:1 M ratio. The result showed the optimal extraction conditions with ChCl-MA (1:1) is 25% water addition, 15 minutes of extraction time, and a 1:20 S/L ratio. These conditions produce total CUR levels of 4.58 mg/g and XNT of 12.93 mg/g; the ethanol 96%-maceration produces 1.88 mg/g total CUR levels and 9.80 mg/g XNT. The most influential variable observed for the extraction was the solid-to-liquid ratio (S/L) and the addition of water (%) (p<0,05). Based on the result, NADES-UAE is more effective than ethanol maceration. The data reported herein are useful for further developments of green extraction methods to extract curcuminoids and xanthorrhizol.

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"Indonesia memiliki keanekaragaman hayati yang tinggi, termasuk kekayaan akanberbagai macam kapang. Salah satu kapang yang bermanfaat bagi kehidupan manusiaadalah Rhizopus. UICC telah mengkoleksi berbagai spesies Rhizopus, tetapi isolat-isolatitu belum semua diteliti sifat-sifat biologinya.Penelitian ini bertujuan untuk mengkaji beberapa sifat biologi tiga isolat kapangRh. microsporus var. chinensis yang meliputi: suhu pertumbuhan optimum; morfologisecara makroskopik dan mikroskopik; pembentukan zigospora; pH pertumbuhanoptimum; aktivitas amilolitik, proteolitik, dan lipolitik yang ditentukan secara kualitatif.Hasil penelitian menunjukkan bahwa Rh. microsporus var. chinensis isolat UICCNo. 16, 18, dan 20 memili.k. i suhu pertumbuhan optimum yang berkisar antara 33° -36°Cpada medium PDA dan TEA; secara makroskopik dan mikroskopik ketiga isolat tidakmemperlihatkan perbedaan bentuk morfologi, walaupun ditumbuhkan pada medium yangberbeda; zigospora tidak terbentuk dari perkawinan ketiga isolat dengan strain-strainRhizopus microsporus dari CBS dan perkawinan antar ketiga isolat itu; pH pertumbuhanoptimum pada medium PDB dan TEB untuk isolat UICC No. 16 dan 18 berkisar antara4,0--6,0, sedangkan isolat UICC No. 20 berkisar antara 4,0--5,0; ketiga isolat kapangmenunjukkan aktivitas amilolitik yang lemah pada medium SA, aktivitas proteolitikditandai dengan kemampuan mencairkan medium NG dalam 2 hari, dan indeks aktivitas.Jipolitik berkisar antara 0,03--0,06 pada medium TA."

"ABSTRAKIndonesia memiliki keanekaragaman mikroorganisme yang tinggi, termasuk kapang tempe, yaitu Rhizopus. Salah satunya adalah Rh. microsporus V. Tiegh. var. rhizopodiformis (Cohn) Schipper & Stalpers. Agar dapat dimanfaatkan lebih lanjut dalam bidang industri, maka perlu diteliti sifat-sifat biologi kapang tersebut. Penelitian ini bertujuan untuk mengkaji beberapa sifat biologi Rhizopus microsporus var. rhizopodiformis isolat UICC no. 6, 13 dan 33 yang meliputi: suhu pertumbuhan optimum; morfologi secara makroskopik dan mikroskopik; pembentukan zigospora; pH pertumbuhan optimum; dan uji kualitatif aktivitas amilolitik, lipolitik dan proteolitik. Hasil penelitian menunjukkan bahwa Rh. micro.sporus var. rhizopodiformis memiliki suhu pertumbuhan optimum berkisar antara 330 36°C pada medium PDA dan TEA; secara makroskopik dan mikroskopik ketiga isolat tidak memperlihatkan perbedaan bentuk morfologi baik pada medium PDA dan TEA; serta mampu membentuk zigospora dan memiliki tipe kawin (+). Isolat UICC no. 6 & 13 pada medium PDB dan TEB mempunyai pH pertumbuhan optimum yang berkisar antara antara 5,0-6,0, sedangkan isolat UICC no. 33 berkisar antara 4,0-5,0 pada medium PDB dan TEB. Ketiga isotat UICC tersebut bersifat amilolitik, lipolitik dan proteolitik."

"ABSTRAKGlukoamilase merupakan enzim ekstraseluler yang dapat menghidrolisis pati nrenjadi glukosa.Penelitian ini bertujuan untuk nengetahui pengaruh empat sumber karbohidrat, yaitu tepung beras, tepung maizena, tepung tapioka, dan soluble starch terhadap aktivitas glukoanilase R. arrhizus UICC 2 dan R. oryzae UICC 128 pada kondisi fermentasi yang diberikan dalam waktu inkubasi 24 jam.Data rata-rata aktivitas glukoamilase R. arrhizus UICC 2 dan R. oryzae UICC 128 dalam waktu fermentasi 24 jam yang dinyatakan dalam unit/ml, diperoleh nilai tertinggi dari sumber karbohidrat tepung rnaizena, kemudian diikuti dengan tepung tapioka, soluble starch, dan tepung beras.Hasil perhitungan aktivitas glukoamilase menunjukkan, ada perbedaan aktivitas glukoamilase R. arrhizus UICC 2 antara sumber karbohidrat tepung maizena dan tepung beras, antara tepung tapioka dan tepung beras, serta antara soluble starch dan tepung beras. Ada perbedaan aktivitas glukoamilase R. oryzae UICC 128 antara tepung maizena dan tepung beras, antara tepung tapioka dan tepung beras, antara soluble starch dan tepung beras, antara tepung maizena dan tepung tapioka, serta antara tepung maizena dan soluble starch.ABSTRACT"