Garuda - Garba Rujukan Digital

Eksplorasi dan karakterisasi bakteri aerob ligninolitik serta aplikasinya untuk pengomposan tandan kosong kelapa sawit Exploration and characterization of ligninolytic aerobic bacteria and its application in composting oil palm empty fruit bunch Haryo Tejo PRAKOSO; Happy WIDIASTUTI; . SUHARYANTO; . SISWANTO
E-Journal Menara Perkebunan Vol 82, No 1: Juni 2014
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstractLignin is a complex compounds that makes up the cell walls of plants and is quite difficult to degrade at normal ambient condition. One of the organic materials with high lignin content is empty fruit bunches (EFB) of oil palm. So far, the well-studied microorganism to degrade lignin is of a class of fungi. Utilization of bacteria to degrade lignin in EFB has rarely been reported although application of the bacteria is very important if it is associated with aerobic composting which requires regular turning process and supporting clean development mechanism (CDM). The objective of this study was to explore and characterize the bacteria having capability to degrade lignin in EFB. The result showed that from 14 types of sample, 12 and 11 isolates were obtained through non enrichment and enrichment methods respectively. Qualitative test was performed using a lignin derivative dye (methylene blue/MB) suspended in Luria Bertani (LB) solid media and the formation of the clear zone was observed, while quantitative assay was performed with enzyme activity assays of laccase (Lac), manganese peroxidase (Mn-P), and lignin peroxidase (Li-P). The best isolate (FS isolate) was obtained from enrichment method that able to make 0.6 cm clear zone of LB media + MB and actively produced laccase, manganese peroxidase with and without addition of Mn with an activity of 2.68, 20.02, and 0.36 U/mL, respectively. While the best isolate from non enrichment method was CRK 1, that was able to make 0.3 cm clear zone and produced Mn-peroxidase with and without addition of Mn as much as 2.09 and 0.23 U/mL, respectively. Application of the decomposer formula could speed upthe declining rate of C/N ratio and suppressing Escherichia coli and Salmonella sp.in EFB compost produced. Abstrak Lignin merupakan senyawa kompleks yang menyusun dinding sel tanaman dan cukup sulit didegradasi secara alami. Salah satu bahan organik yang mempunyai kadar lignin tinggi adalah tandan kosong kelapa sawit (TKKS). Sejauh ini, mikroorganisme yang banyak dipelajari dalam mendegradasi lignin adalah dari golongan jamur. Peng-gunaan bakteri dalam mendegradasi lignin pada TKKS belum banyak dilaporkan walaupun peran bakteri lignino-litik aerob sangat penting jika dikaitkan dengan proses pengomposan secara aerob yang membutuhkan pembalikan secara berkala danprogram clean development mechanism (CDM). Penelitian ini bertujuan mengeksplorasi dan meng-karakterisasi bakteri yang berpotensi mendegradasi lignin dalam pengomposan TKKS. Dari 14 jenis sampel diperoleh sebanyak 12 dan 11 isolat melalui metode tanpa dan dengan pengkayaan. Uji kualitatif dilakukan dengan mengukur terbentuknya zona bening pada media Luria Bertani (LB) padat yang mengandung senyawa warna turunan lignin (biru metilen/MB).Uji kuantitatif dilakukan dengan mengukur aktivitaslakase, Mn-peroksidase, dan lignin peroksidase. Hasil penelitian menunjukkan bahwa isolat FS merupakan isolat terbaik dari metode pengkayaan yang mampu membentuk zona bening pada medium LB + MB 0,6 cm, sedangkan isolat terbaik dari metode tanpa pengkayaan adalah CRK 1 dengan zona bening 0,3 cm pada medium yang sama setelah inkubasisemalam. Isolat FS memiliki aktivitas lakase, Mn-peroksidase dengan dan tanpa Mn berturut-turut adalah sebesar 2,68; 20,02; dan0,36 U/mL, sedangkan isolat CRK 1 memiliki aktivitas Mn-peroksidase dengan dan tanpa Mnberturut-turut adalah 2,09 dan 0,23 U/mL. Aplikasi formula dekomposer pada pengompos-an 200 ton TKKS mampu mempercepat laju penurunan nisbah C/N dan menekan populasi Escherichia coli dan Salmonella sp.

Solubilization of insoluble phosphates by Aspergillus niger Pelarutan fosfat sukar larut oleh Aspergillus niger LAKSMITA P SANTI; D H GOENADI; . SISWANTO; I SAILAH; . ISROI
E-Journal Menara Perkebunan Vol 68, No 2: Desember 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

RingkasanPenggunaan langsung fosfat alam (FA) ke dalam tanah sebagai sumber pupuk P telah dilakukan selama bertahun-tahun melalui beberapa macam cara penggunaan. Kualitas FA di Indonesia umumnya rendah dan ketersediaan bahan baku yang berkualitas untuk produksi pupuk fosfat terlarut relatif terbatas. Beberapa mikroba asal tanah yang dapat melarutkan fosfat anorganik telah banyak dilaporkan. Namun, informasi yang tersedia tentang mekanisme pelarutan P dari FA lokal asal Indonesia dan P anorganik oleh Aspergillus niger BCC F194 belum banyak diteliti. Satu seri penelitian laboratorium telah dilaksanakan untuk mengetahui kemampuan A. niger BCC F194 melarutkan P. Evaluasi agronomi FA lokal (FA Cileungsi dan Madura) di rumah kaca juga telah dilakukan. A. niger BCC F194 dapat melarutkan sumber P sukar larut, yaitu FA Cileungsi dan Madura, serta senyawa Ca3(PO4)2 dan AlPO4. Kelarutan P anorganik tersebut berhubungan dengan peningkatan aktivitas proton (H*) yang menyebabkan penurunan pH medium dan produksi asam organik. Asam organik utama yang dihasilkan oleh A. niger BCC F194 dalam medium cair Pikovskaya yang dimodifikasi adalah asam oksalat (3.75 mM), asam sitrat (2.0 mM), dan asam glukonat (0.9 mM). Kelarutan FA Cileungsi lebih besar dibandingkan dengan FA Madura, dan kelarutan Ca3(PO4)2 lebih besar dibandingkan kelarutan AlPO4. Tidak ada korelasi antara kelarutan P anorganik dengan aktivitas enzim fosfatase, walaupun aktivitas enzim fosfatase cukup tinggi terdeteksi dalam medium. Satu formula biosuperfosfat telah berhasil dirakit dengan mereaksikan FA lokal dengan supernatan kultur cair (SKC) pengganti asam sulfat. Hasil percobaan padabibit kakao, karet dan kelapa sawit di rumah kaca menunjukkan bahwa prototipe pupuk biosuperfosfat dengan bahanbaku FA Cileungsi dan Madura bentuk granul maupun serbuk, memiliki nilai efektivitas agronomi yang relatif menyamai SP-konvensional. SummaryThe direct application of rock phosphate (RP) to soils as a source of phosphorus (P) fertilizer has been employed with varying popular methods over the years. The RP in Indonesia has low quality for plant fertilization and the availability of the raw material with good quality for production of soluble phosphate fertilizers is limited. Many common soil microbes that can dissolve insoluble inorganic phosphate have been extensively studied. However, there is little information on mechanism of P-solubilization from local RP of Indonesia and inorganic P by Aspergillus niger BCC F194 isolated from tropical acid soils. A laboratory study was conducted to determine the ability of phosphate solubilization A. niger BCC F194. Agronomic evaluation of bioactivated local RP, i.e. Cileungsi and Madura phosphate rocks (CRP and MRP) for direct application in greenhouse experiment was also conducted. A. niger BCC F194 was able in solubilizing different types of hardly-soluble phosphates, i.e. CRP and MRP, Ca3(PO4)2, and AlPO4 compounds. Inorganic P solubilization was directly correlated to the organic acid production and increasing proton (H+) activities causing pH to decrease and production of organic acid. The major acidic metabolites produced by A. niger BCC F 194 in modified liquid culture Pikovskaya medium were oxalic acid (3.75 mM), citric acid (2.0 mM), and gluconic acid (0.9 mM). The solubilization of Cileungsi RP was higher than that of Madura RP, and the solubilization of Ca3(PO4)2 was better than that of AlPO4. No correlation between solubilization of inorganic P and enzyme activities, although high activities of phosphatase enzyme were detectable in the medium. A biosuperphosphate formula had been constructed by reacting local RP with liquid culture supernatant (LCS) replacing sulfuric acid. Results of cocoa, rubber, and oil palm seedlings experiments in greenhouse indicate that both granular and powder biosuperphosphate prototypes commonly had a comparable relative agronomic effectiveness value to that of the conventional SP.

Penggunaan enzim protease pada pengolahan lateks pekat DPNR sebagai bahan pembuatan sphygmomanometer Use of protease on the processing of concentrated latex DPNR as material for sphygmomanometer manufacturing . SISWANTO; . SUHARYANTO; Yoharmus SYAMSU
E-Journal Menara Perkebunan Vol 77, No 2: Desember 2009
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstractIn order to increase competitiveness in the international market especially in USA, the domestic industrial manufactures of latex dipping products have to meet the FDA requirement for protein standard that is 150 g protein/g. Use of cheap protease from an effective local sources will support the production of concentrated latex with low protein so that the end product will meet FDA prerequisite of standard protein. Local source of proteases from Bacillus sp. isolated from latex coagula serum (LCS), papain and bromeline were examinated their proteolytic activity using casein and casein mixed with LCS (1:1) as substrate. The best protease source will be applied to produce deproteinized natural rubber (DPNR) of concentrated latex, and furthermore used as raw material in producing sphygmomanometer at commercial scale. The objective of this research is to determine the best protease source and condition of optimum activity and its effectiveness for producing DPNR of concentrated latex as raw material for sphygmomanometer production. The result showed that Bacillus sp. K3 is the best isolate for protease producer with protease activity of 0.438 U/mL under room temperature (28-30oC) for three days. Of three sources of protease tested, papain was the most active one when casein was used as substrate. The used of LCS as substrate was not efficient because of the presence of protease inhibitor which could not be removed by heating at 100C for five minutes. The proteolytic activity of papain was optimum at room temperature 37C and pH 7.7-11 i.e achieved 0.6-0.7 U/mL. Sphygmomanometers component produced by concentrated latex non DPNR containing 0,27-0.31% total N and 445-710 g extractable protein/g, whereas sphygmo-manometers component produced by latex DPNR containing 0.18-0.28% total N and 79-103 extractable protein thus pass its protein content prerequisite of FDA (<150 g /g). Sphygmo-manometers component produced by con-centrated latex DPNR have physical properties such as tensile strength, modulus 300% and elongation at break better than conventional concentrated latex.AbstrakUntuk meningkatkan daya saing di pasar internasional khususnya Amerika Serikat, barang celup lateks alam produksi dalam negeri harus memenuhi standar protein yang ditetapkan oleh FDA yaitu 150 g protein/g. Penggunaan enzim protease dari sumber lokal yang murah dan efektif akan membantu dalam pembuatan lateks pekat rendah protein sehingga produk yang dihasilkan memenuhi standar protein yang disyaratkan FDA. Sumber enzim protease lokaldari isolat Bacillus sp. yang diisolasi dari serum bekuan lateks (SBL), papain dan bromelin diuji aktivitas proteo-litiknya dengan substrat kasein dan campuran kasein dan SBL (1:1). Sumber enzim protease terbaik digunakan untuk produksi lateks pekat deproteinized natural rubber (DPNR) dan selanjutnya lateks tersebut digunakan untuk percobaan produksi komponen sphygmomanometer skala komersial. Penelitian bertujuan menetapkan sumber protease terbaik dan kondisi optimum aktivitasnya untuk pembuatan lateks pekat DPNR dan komponen sphygmomanometer. Hasil penelitian menunjukkan bahwa Bacillus sp. K3 adalah isolat terbaik dalam menghasilkan enzim protease yaitu mencapai 0,438 U/mL pada inkubasi suhu ruang (28-30oC) selama tiga hari. Dari ketiga sumber protease yang diuji, enzim papain menujukkan aktivitas terbaik ketika diuji dengan substrat kasein. Penggunaan subtrat SBL kurang sesuai untuk produksi protease karena adanya inhibit orprotease yang tidak bisa dihilangkan dengan cara pemanasan pada suhu 100C selama lima menit. Aktivitas enzim papain optimum pada suhu 37C dan antara pH 7,7-11, yaitu mencapai 0,6- 0,7 U/mL. Komponen sphygmomanometer konvensional yang dibuat dengan bahan baku lateks pekat non DPNR memiliki kadar N total 0,27-0,31% dan kadar protein terekstrak 445- 710 g/g, sedangkan komponen sphygmomanometer yang diproduksi dengan lateks pekat DPNR memiliki kadar N total 0,18-0,28% dan protein terekstrak 79-103 g/g sehingga memenuhi ambang batas yang ditetapkan oleh FDA yaitu <150 μg/g. Sifat fisika seperti tegangan putus, modulus 300%, dan perpanjangan putus komponen sphygmomanometer yang dibuat dari lateks pekat DPNR lebih baik dari pada lateks pekat non DPNR.

Transformation of Coffee arabica using chitinase gene and regeneration of planlets from transformed-zygotic embryos Transformasi Coffea arabica menggunakan gen kitinase dan regenerasi planlet dari embrio zigotik-transforman Asmini BUDIANI; T CHAIDAMSARI; . PRIYONO; S MAWARDI; . SISWANTO
E-Journal Menara Perkebunan Vol 68, No 2: Desember 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

RingkasanRekayasa genetika kopi arabika tahan penyakit cendawan dapat dilakukan dengan cara memasukkan gen kitinase (gen chi) ke dalam genom tanaman tersebut. Penelitian ini bertujuan untuk mengintroduksikan gen chi pada kopi arabika serta meregenerasi eksplan yang ditransformasi menjadi plantlet. Gen chi disubkloning dari pBS G11 ke dalam plasmid pCAMBIA2301. Melalui Agrobacterium tumefaciens, plasmid rekombinan pCAMBIA2301/35s-chi kemudian dimasukkan ke dalam eksplan daun dan embrio zigotik kopi arabika. Eksplan daun transforman ditumbuhkan pada media seleksi yang mengandung kanamisin untuk induksi kalus embriogenik . Beberapa kombinasi 2,4-D dan dicamba serta kinetin, BAP dan 2-iP diuji kemampuannya untuk menginduksi terbentuknya kalus embriogenik. Embrio zigotik transforman ditumbuhkan pada media MS modifikasi yang mengandung kanamisin. Hasil penelitian menunjukkan bahwa perbedaan tipe sitokinin dan kombinasinya dengan 2,4-D atau dicamba. menyebabkan terjadinya variasi persentase pembentukan kalus embriogenik tahan kanamisin. Penambahan 100 mg/L kanamisin dalam media seleksi cukup efektif untuk menghambat pertumbuhan eksplan daun nontransforman. Persentase tertinggi induksi kalus embriogenik pada eksplan daun non transforman maupun transforman diperoleh pada media yang mengandung 5 mM 2,4- D dengan 5 mM of kinetin atau 5 mg/L dicamba dengan 5 mM BAP. Sedangkan dalam media dengan penambahan 5 mM kinetin, 100 mg/L asam sitrat dan 100 ppm asam askorbat, jumlah eksplan yang membentuk kalus mencapai optimum pada konsentrasi 0 dan 1 ppm dicamba untuk eksplan transforman dan 10 mg/L dicamba untuk non transforman. Pada eksplan embrio zigotik transforman, peningkatan konsentrasi kanamisin dari 100 mg/L hingga 500 ppm menurunkan persentase pengecambahan embrio dari 80.5 % menjadi 49%, persentase perakaran, dari 34 % menjadi 16%, jumlah akar, panjang akar dan tinggi tunas dari 7 mm menjadi 4 mm. Pada semua perlakuan kanamisin, embrio zigotik non transforman tidak membentuk akar dan pada umur kultur yang sama tunas yang dihasilkan lebih pendek dibandingkan dengan embrio-zigotik transforman. Hasil tersebut membuktikan bahwa gen ketahanan terhadap kanamisin (NPTII) telah terinsersi dan terekspresi dengan baik pada plantlet kopi arabika yang berasal dari eksplan embrio-zigotik transforman. Karena gen chi dikonstruksi dalam satu vektor dengan NPTII, maka diharapkan gen tersebut juga telah terinsersi ke dalam genom tanaman kopi.SummaryGenetic engineering of arabica coffee resistant to fungal diseases might be done by introducing a chitinase-encoding gene (chi) into genome of this plant. This research was aimed to introduce chi construct into arabica coffee and regenerate plantlets from the transformed explants. The chi gene was previously subcloned from pBS G11 into pCAMBIA2301 plasmid. With Agrobacterium tumefaciens,the recombinant plasmid pCAMBIA2301/35s-chi was then introduced into leaf and zygotic embryos explants of arabica coffee. The transformed leaf explants were cultured on the selection media containing kanamycin in the presence of several combinations of 2,4-D and dicamba with kinetin, BAP and 2-iP to induce the formation of embryogenic callus. The transformed zygotic embryos were cultured on the media of modified MS containing kanamycin. The results showed that the several types of cytokinin used in combination with 2,4-D or dicamba caused the percentage of kanamycin resistant-embryogenic calli was varied. The addition of 100 mg/L kanamycin in the selection media was effective for inhibiting the growth of untransformed explants. Among the several combinations of auxin and cytokinin tested, the highest percentage of embryogenisis for untransformed and transformed leaf explants were achieved on the media containing 5 mM 2,4-D and 5 mM kinetin or 5 mg/L dicamba and 5 mM BAP. However in the presence of 5 mM kinetin together with antioxidants of 100 mg/L citric acid and 100 mg/L ascorbic acid, the explants calluses was optimum at 0 - 1 mg/L dicamba for transformed explants and 10 mg/L dicamba for untransformed explants. In the explants of transformed-zygotic embryos, increasing kanamycin from 100 mg/L up to 500 mg/L decreases the percentage of embryo germination from 80.5 % to 49%, rooted-shoots from 34 % to 16%, number of roots, root length and shoot length from 7 mm to 4 mm. At all the kanamycin treatments, root was not developed from the untransformed-zygotic embryos and the lenght of shoots were shorter compared to the transformed-zygotic embryos. This result demonstrates that the kanamycin-resistant gene (NPTII) has been inserted and well expressed in the plantlets of arabica coffee derived from transformed-zygotic embryos. Since the chi gene was constructed in one vector with NPTII, this gene might also been inserted in the genome of coffee.

Produksi imunoglobulin Y (IgY) untuk pengembangan metode deteksi dini kontaminasi okratoksin (Immunoglobulin Y (IgY) production to develop an early detection method for ochratoxin contamination) Irma KRESNAWATY; . SUHARYANTO; . SISWANTO; Sumi HUDIYONO
E-Journal Menara Perkebunan Vol 86, No 1 (2018): April, 2018
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v1i1.279

Indonesian coffee and cocoa commodities are constrained by low product quality problem due to contamination of fungal metabolites which producing ochratoxin A (OTA). Ochratoxin is neprotoxic, immunogenic, carcinogenic and teratogenic to the human health. Early detection method on site detection should be developed because of those negative effects. The aim of this study was to produce antibody to develop a method for OTA detection. Antibody was produced by immunization of egg laying hen. Antibody-produced was sepatared and analyzed using ELISA (Enzyme-Linked Immunosorbent Assay) and DBIA (dot blot immunoassay),and tested its composition using HPLC and SDS PAGE. The results showed that anti-OTA polyclonal antibodies had been obtained already from chicken eggs in the 4th period (7 weeks after initial immunization). These antibodies showed anti-OTA reactivity by DBIA method and still showed anti-OTA reactivity up to 9th period (12 weeks after initial immunization). The anti-BSA antibodies produced should be removed to increase the sensitivity of antibodies againts ochratoxin A. The separation of BSA antibodies can be conducted by the absorption of the protein. [Keywords: ochratoxin A; early detection; antibody IgY]. AbstrakKomoditas kopi dan kakao Indonesia terkendala masalah mutu produk yang rendah akibat kontaminasi cendawan penghasil okratoksin A. Okratoksin A (OTA) bersifat neprotoksik, imunogenik, karsinogenik dan teratogenik yang membahayakan kesehatan manusia. Karena efek negatif yang diakibatkan oleh mikotoksin ini, maka perlu dikembangkan deteksi dini kontaminasi okratoksin langsung di lokasi. Penelitian ini bertujuan menghasilkan antibodi imunoglobulin Y (IgY) untuk mengembangkan metode perakitan perangkat deteksi cepat berbasis imunologi untuk deteksi OTA. Antibodi dihasilkan menggunakan uji ayam petelur. Antibodi yang dihasilkan dipisahkan dan dianalisis aktivitasnya dengan ELISA (Enzyme-Linked Immunosorbent Assay) dan DBIA (dot blot immunoassay), serta diuji komposisinya dengan HPLC dan SDS PAGE. Hasil penelitian menunjukkan bahwa antibodi poliklonal anti-OTA sudah diperoleh dari telur ayam pada periode ke-4 (7 minggu setelah imunisasi awal). Antibodi ini menunjukkan reaktivitas anti-OTA dengan metode DBIA dan masih menunjukkan reaktivitas anti-OTA sampai periode 9 (12 minggu setelah imunisasi awal). Komposisi asam amino antibodi anti-OTA menunjukkan perbedaan dengan komposisi asam amino IgY di database. Antibodi anti BSA yang dihasilkan harus dihilangkan terlebih dahulu untuk meningkatkan sensitivitas antibodi terhadap okratoksin A dan pemisahan dapat dilakukan dengan penyerapan antibodi BSA.[Kata Kunci: okratoksin A; deteksi dini; antibodi IgY].

Immuno-chemiluminescense detection of allergenic proteins from rubber gloves and natural rubber latex Deteksi protein allergen secara imunokimia dari sarung tangan dan lateks karet alam . Siswanto
E-Journal Menara Perkebunan Vol 83, No 1: Juni 2015
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstrakLateks alam maupun produk jadi yang berasal dari karet alam diketahui mengandung protein alergen. Namun demikian identifikasi jenis protein allergen belum banyak dilaporkan. Penelitian ini bertujuan untuk mendeteksi protein alergen dari sarung tangan dan lateks karet alam menggunakan metode immuno-chemiluminescense. Protein di-ekstrak dari tiga fraksi sentrifugasi lateks (serum B, serum C dan partikel karet) serta tujuh jenis sarung tangan komersial, kemudian dipisahkan berdasarkan berat molekulnya melalui Gel elektroforesis 1-D (SDS PAGE) dan 2-D. Selanjutnya untuk deteksi protein allergen secara immuno-chemiluminescense dilaku-kan imunobloting menggunakan serum Ig_E tiga pasien yang terbukti positif alergi terhadap protein asal sarung tangan lateks, kemudian diwarnai dengan Sypro Ruby protein blot fluorescence. Hasil penelitian menunjukkan bahwa berdasarkan hasil analisis Western blot one-DE sampel protein lateks menggunakan serum tiga orang tenaga medis yang terbukti positif alergi terhadap protein lateks, maka dapat diidentifikasi 14 jenis protein alergen pada sarung tangan lateks, empat diantaranya merupakan pita major yaitu Berat Molekul (BM) 35, 38, 46 dan 56 kDa. Protein allergen pada sarung tangan tersebut kemungkinan berasal dari bagian C-serum terutama protein BM 46 dan 56 kDa ataupun campuran antara C-serum dan B-serum dari lateks karet alam. Hal ini dibuktikan bahwa dari sampel C-serum lateks dapat teridentifikasi 12 protein alergen, empat diantaranya merupakan pita major yaitu BM 42, 46, 51 dan 56 kDa. Sedangkan dari sampel B-serum teridenti-fikasi tiga pita major dengan BM 14, 16 and 51 kDa. Hasil analisis Western blot 2-DE ekstrak protein sarung tangan menggunakan serum tiga orang tenaga medis yang terbukti positif alergi terhadap protein lateks, maka dapat diidentifikasi 12 - 13 spot protein alergen dengan pI at 4.0 to 7.0 dan yang paling dominan adalah dengan BM 23, 35, 38, 42, 45, 46 kDa.Abstract Natural rubber latex and finished products derived from natural rubber is known to contain allergenic proteins. Nevertheless identification of allergenic protein has not been widely reported. This study aims to detect the protein allergens from the glove of hands and natural rubber latex using immuno-chemiluminescense. Proteins extracted from the latex centrifugation three fractions (serum B, serum C and rubber particles) as well as seven types of commercial gloves, then separated by molecular weight through 1-D gel electrophoresis (SDS PAGE) and 2-D. Furthermore, for the detection of allergen proteins in immuno-chemiluminescense performed immunoblotting using the serum IgE three patients who tested positive for allergy to latex gloves native protein, and then stained with fluorescence Sypro Ruby protein blot. The results showed that based on the results of Western blot analysis of one-DE latex proteins using serum samples three medical personnels who tested positive for allergy to latex proteins, we can identify 14 types of protein allergens in latex gloves, four of which are major bands that having Molecular Weight (MW) 35, 38, 46 and 56 kDa. Protein allergen on the gloves are likely to come from the C-serum protein mainly MW 46 and 56 kDa, or a mixture of C-serum and B-serum of natural rubber latex. It was proved that from C-serum samples could be identified as many as 12 protein latex allergens, four of which were major bands that MW 42, 46, 51 and 56 kDa. While the B-serum samples identified three major bands with MW 14, 16 and 51 kDa. Results of Western blot analysis of 2-DE protein extracts glove using the serum three medical personnel who tested positive for allergy to latex proteins, it could be identified 12-13 allergen protein spot with pI at 4.0 to 7.0 and most dominant is the MW 23, 35, 38, 42, 45, 46kDa.

Optimasi pembuatan membran chitosam dalam penurunan COD dan BOD POME (Palm Oil Mill Effluent) [Optimization of the membrane production process to COD and BOD removal of POME (Palm Oil Mill Effluent)] Sri WAHYUNI; . SISWANTO; Alia DAMAYANTI
E-Journal Menara Perkebunan Vol 84, No 1: Oktober 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Palm oil is one of the main commodities that cultivated in Indonesia as the biggest palm oil producer. One of the main problems in palm oil industry is the difficulty to degradate the palm oil mill effluent (POME) due to the high quantity and content of COD and BOD. In physico-chemical, POME can be processed using membrane filtration technology. Chitosan is one of the most widely used material forproducing membrane filtration. Composite of Chitosan-PVA-PEG is a highly mixture absorbent, which possibly can be used as a membrane in filtration process of POME. The experiment was started with the production of composite membrane, and then filtration application using cross-flow reactor system. The variables of this experiment were chitosan and PVA ratio (3:7, 4:5, 1:1, 6:4 and 7:3 (v/v)), and stirring speed (100 rpm and 300 rpm). The reactor test was conducted for 50 minutes and permeate were taken every 10 minutes. Filtration output parameters that were analyzed flux, COD and BOD. The result showed that the highest flux values in the variation of the stirring speed of 100 rpm and 300 rpm were 40.20 L/m2.hr and 27.15 L/m2.hr, respectively. The highest rejection values of COD and BOD were obtained in membrane ratio variation of 1:1 (v/v) and stirring speed of 300 rpm, which are 97.24% and 97.60%, respectitively.

Overexpression of chitinase gene with a GC-rich synthetic enhancer in tobacco plant (Nicotiana tabacum L.) Overekspresi gen kitinase dengan enhancer sintetis kaya GC pada tanaman tembakau (Nicotiana tabacum L.) D SANTOSO; T CHAIDAMSARI; A BUDIANI; H MINARSIH; S DWI UTOMO; . SISWANTO
E-Journal Menara Perkebunan Vol 68, No 2: Desember 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Ringkasan Perakitan tanaman perkebunan toleran terhadap serangan cendawan patogenik dilakukan dengan mengoverekspresikan gen penyandi kitinase. Untuk itu elemen DNA peningkat ekspresi (enhancer E52) yang berupa oligonukleotida 52 pb dan kaya kandungan basa purin (GC) disisipkan di ujung 5’ konstruk 35S-chi. Penyisipan E52 tersebut dilakukan secara lebih terarah pada situs ganda HindIII-SalI dari MCS pCAMBIA2301. Melalui situs HindIII yang terletak tepat di ujung 3’ E52, konstruk 35S-chi kemudian disambungkan dengan E52 pada pCAMBIA tersebut. Transformasi DNA rekombinan ke dalam sel tembakau dikerjakan melalui perantaraan Agrobacterium tumefaciens LBA4404. Sel tanaman transgenik diseleksi dan diregenrasi dalam media yang mengandung 3% sukrosa, 0,5 mg/L diregenerasikan pada media MS padat benzilaminopurin (BAP) dan 50 mg/L kanamisin. Pada media ini tunas transgenik tembakau mulai terbentuk setelah 5 minggu penanaman. Analisis tingkat aktivitas enzimatis menunjukkan bahwa aktivitas kitinase pada tembakau transgenik 40 hingga 80 kali lebih tinggi daripada non-transgenik. Pengujian hibridisasi protein menggunakan antibodi anti-kitinase, dot blot dan western blot, membuktikan bahwa enhancer tersebut dapat meningkatkan ekspresi transgen chi pada tanaman tembakau.Summary Development of estate crops tolerant to pathogenic fungi is conducted by overexpressing chi gene. For this purpose, a synthetic enhancer consisting of 52 base pairs and GC-rich was inserted at immediate 5’ end of a 35S-chi cassette. Insertion of the E52 was directed at HindIII-SalI restriction sites of the pCAMBIA2301 MCS. With HindIII restriction site located just after the 3’ end of the E52 sequence, the 35S-chi construct was then ligated with the E52 of the pCAMBIA. Transformation of the resulting recombinant DNA into tobacco cells was mediated by Agrobacterium tumefaciens LBA4404. The transgenic cells were selected and regenerated on a solid MS medium supplemented with 3% sucrose, 0.5 mg/L benzylamino purine (BAP) and 50 mg/L kanamycin. Tobacco shoots were initiated after 5 weeks inoculation on the selection media. Enzymatic analysis demonstrated that chitinase activity of transgenic tobacco was 40 to 80 folds higher than that of the control plant. Analysis of enzymatic activity using hybridization with anti-chitinase antibody indicated that the level of chitinase activity in the transgenic tobacco carrying the enhancer is higher than that without enhancer. These data suggest that the enhancer improved the expression of chi transgene in tobacco.

Optimisasi produksi biogas dari limbah lateks cair pekat dengan penambahan logam Optimization of biogas production from concentrated-latex effluent with addition of metals Irma KRESNAWATY; I SUSANTI; . SISWANTO; . TRI-PANJI
E-Journal Menara Perkebunan Vol 76, No 1: Juni 2008
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Summary The treatment of concentrated-latex effluent process applied in the field presently, has not obtain optimum additional benefits. Besides that, the technology using ponding system needs wide area and causes air pollution that such a way caused conflicts with society. The application concept of clean industry: reuse, reduction, recovery and recycling, makes the possibilities to convert the effluent to be usefull products. One of the alternative effluent process is by utilizing it as the source of renewable energy, that is in the form of biogas as an alternative energy. The preliminary research showed that the use of spontaneous latex skim coagulation, the addition of 1% manure as source of seed, and leaf biomass as the source of carbon could increase the biogas production. This research was carried out to optimize biogas production by adding metal ion and to observe the parameters which influenced every stage of biogas production. At the beginning of the process, pH showed increasing due to the hydrolysis process that generally occured in acid condition, but it remained stable (6.6-7.7) in the next steps, whereas, the VFA value as well as BOD value tended to increase. COD value had fluctuative inclination caused by the conversion of organic compounds to produce biogas and the hydrolysis process of leaf biomass to organic compounds that decom-posed to further biogas. The best result of biogas production was showed by addition of Fe3+ with optimum concentration 0.50 mg/L effluent.

Optimasi pertumbuhan dan aktivitas enzim ligninolitik Omphalina sp. dan Pleurotus ostreatus pada fermentasi padat Optimization of growth and ligninolytic enzymes activity of Omphalina sp. and Pleurotus ostreatus using solid state fermentation Happy WIDIASTUTI; . SISWANTO; . SUHARYANTO
E-Journal Menara Perkebunan Vol 75, No 2: Desember 2007
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

SummarySolid wastes of bagasse and empty fruitbunch (EFB) respectively from sugarcane andpalm oil mill in Indonesia are abundant. Nowdays, up to now these solid wastes have not yetbeen optimally utilized so that the added valueis still very low and even cause an environ-mental problem. Research on bioconversion ofbagasse and EFB with two culture of white-rotfungi (WRF) i.e., Omphalina sp. and Pleurotusostreatus to produce ligninolytic enzymes wasconducted to provide added value to thislignocellulosic waste. Production of extracellular enzymes from WRF was not onlydetermined by the type of isolate but also theculture condition. This research was aimed todetermine the optimum culture condition ofsolid state fermentation in producing lignino-lytic enzymes at laboratory scale. In thisresearch, WRF was examined for ligninolyticproducing enzymes (laccase, lignin peroxidase/ LiP and Mn-peroxidase / MnP), using mediaconsisting of bagasse and EFB separately asmain substrate with supplementation of ricebran, Cu 2+ with or without rice bran. Theobservation was based on their growth andligninolytic enzyme activities. Characteristicsof optimum pH of LiP, MnP and laccaseactivity were also determined. The resultsshowed that addition of supplement was notable to increase the Cu 2+ growth of myceliaespecially in the first and second months but inthe third month the addition of supplementenhanced the mycelia growth. The growth ofmycelia on the addition of Cu 2+ with or withoutrice bran significantly lower compared to thecontrols both of Omphalina sp. and P. ostreatusin bagasse and EFB. The optimum pH oflaccase, MnP, and LiP activities was five bothfor Omphalina sp. and P. ostreatus at EFBand bagasse. Omphalina sp. was better thanP. ostreatus in producing laccase on bagasseand EFB without any supplementations. Thehighest laccase activity showed by P. ostreatuswith bagasse and EFB media treated with Cuand Cu + rice bran. Supplementation withCu 2+ was more effective in increasing laccaseactivity than rice bran. Activities of Li-P onbagasse and EFB for the two WRF cultureswere significantly influenced by supple-mentation of both of rice bran and Cu 2+ . Li-Pactivity on EFB was slightly higher than thaton bagasse. Mn-P activity was not influencedby rice bran, Cu 2+ or the combination of both.However, these enzymes activities on EFBwere higher compared to bagasse especiallyfor P. ostreatus. Suplementation of Cu wasenhance the activity of laccase and LiP both ofP. ostreatus and Omphalina sp in baggasse andEFB though inhibited the growth of those fungiespecially in the initial growth.RingkasanLimbah padat bagas tebu dan tandankosong kelapa sawit (TKKS) masing-masingdari proses pengolahan gula tebu dan minyaksawit di Indonesia jumlahnya melimpah dansampai saat ini belum mendapat penangananyang efektif sehingga nilai tambahnya masihsangat rendah dan bahkan mengganggulingkungan. Penelitian biokonversi limbahpadat bagas tebu dan TKKS menggunakan duaisolat fungi pelapuk putih (FPP) yaituOmphalina sp. dan Pleurotus ostreatus untukproduksi enzim ligninolitik dilakukan untukmeningkatkan nilai tambah limbah ligno-selulosa tersebut. Penelitian ini, mengujiaktivitas enzim ekstraseluler dari FPP antaralain lakase, lignin peroksidase (LiP), dan Mn-peroksidase (MnP) dari dua spesies FPP yaituOmphalina sp. dan P. ostreatus. Penelitianbertujuan menetapkan kondisi optimum mediafermentasi untuk produksi enzim ligninolitikdari bagas tebu dan TKKS sebagai substrat dankarakterisasi pH optimum enzim ligninolitikdari dua FPP yaitu Omphalina sp. danP. ostreatus. Pengamatan dilakukan berdasar-kan laju pertumbuhan dan aktivitas enzimligninolitik. Enzim lakase, MnP, dan LiPdiekstraksi dan dikarakterisasi pH optimumaktivitasnya. Hasil penelitian menunjukkanbahwa pemberian suplemen menghambatpertumbuhan miselia pada satu dan dua bulanpertama inkubasi, namun laju pertumbuhanmiselium khususnya pada perlakuan pemberianCu 2+ dan Cu 2+ + dedak meningkat tajam padabulan ketiga setelah inkubasi. Pertumbuhanmiselium Omphalina sp dan P. ostreatus padamedium yang ditambah Cu 2+ dan Cu 2+ +dedaklebih rendah dibandingkan dengan kontrol.Pada inkubasi tiga bulan, aktivitas optimumlakase, MnP dan LiP diperoleh pada pH 5, baikuntuk Omphalina sp. maupun P. ostreatusyang diekstrak dari bahan lignoselulosa bagastebu dan TKKS. Aktivitas lakase dariOmphalina sp. lebih tinggi daripadaP. ostreatus pada substrat TKKS dan bagastebu tanpa suplementasi. Pemberian suplemenberupa Cu 2+ dan dedak atau kombinasinyameningkatkan aktivitas lakase baik pada bagastebu maupun pada TKKS. Aktivitas lakasetertinggi ditunjukkan oleh isolat P. ostreatuspada medium bagas tebu dan TKKS padaperlakuan pemberian Cu 2+ dengan atau tanpadedak. Aktivitas lakase nampaknya lebihdipengaruhi oleh penambahan Cu 2+ dibanding-kan dengan pemberian dedak. Aktivitas LiPbaik pada bagas tebu maupun TKKS untukkedua FPP yang diuji pada perlakuanpenambahan dedak dan Cu nyata lebih tinggidibandingkan dengan aktivitas LiP yangdiekstrak dari medium tanpa penambahansuplemen. Aktivitas LiP pada TKKS lebihtinggi dibandingkan dengan pada bagas tebukhususnya untuk P. ostreatus. Sedangkanaktivitas MnP tidak dipengaruhi penambahandedak dan Cu 2+ demikian pula kombinasikeduanya. Aktivitas MnP yang diekstrak dariTKKS lebih tinggi dibandingkan denganaktivitas MnP yang diekstrak dari bagas tebukhususnya untuk P. ostreatus. PenambahanCu 2+ meningkatkan aktivitas lakase dan LiPP. ostreatus dan Omphalina sp yang ditum-buhkan pada bagas dan TKKS walaupun ionlogam ini menghambat pertumbuhan keduaJPP ini khususnya pada awal pertumbuhan.

Title

Found 32 Documents
Search

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Title Search

Found 32 Documents Search

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Title

Found 32 Documents
Search