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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
Menara Perkebunan Vol. 77 No. 2: 77 (2), 2009
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v77i2.82

Abstract

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.
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
Menara Perkebunan Vol. 76 No. 1: 76 (1), 2008
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v76i1.92

Abstract

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.
Ekspresi β -1,3 glukanase dan kitinase pada tanaman kopi arabika (Coffea arabica L.) tahan dan rentan karat daun Expression of β-1,3 glucanase and chitinase of arabica coffee (Coffea arabica L.) resistant and susceptible against leaf rust disease Asmini BUDIANI; I SUSANTI; Surip MAWARDI; D A SANTOSO; . SISWANTO
Menara Perkebunan Vol. 72 No. 2: 72 (2), 2004
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v72i2.122

Abstract

Summary Leaf rust disease caused by Hemileia vastatrix is considered to be one of the most important diseases on arabica coffee plantation. In order to understand the mechanism underlying resistance of arabica coffee against leaf rust disease, this research was aimed to study expression of β-1,3 glucanase (GLU) and chitinase (CHI) genes in the arabica coffee S1934 and  BLP10 that have been reported respectively as a resistant and susceptible varieties to H. vastatrix. The two varieties were essayed against H. vastatrix, and an RT-PCR (Reverse Transcriptase Polymerase Chain Reaction) using total RNAs  from the S1934 and BLP10, both inoculated with H. vastatrix and uninnoculated was carried out for studying the expression of GLU and CHI. Two primer pairs were designed to amplify the conserved region of GLU and CHI. Amplification products were sequenced and the nucleotide sequences were subjected to BlastX analysis. The result of bioassay confirmed that arabica coffee S1934 was resistant to H. vastatrix, while BLP10 was susceptible.   β-1,3 glucanase was expressed in all of the four samples, the inoculated and uninnoculated S1934, and BLP10 in different degree. S1934 expressed higher GLU compared to BLP10. In the inoculated S1934 the expression of this gene was higher compared to that of the uninoculated one. Expression of CHI was detected only in the S1934, both inoculated and uninoculated. Sequence analysis confirmed that the RT-PCR products were exon regions of genes encoding β-1,3 glucanase dan chitinase respectively. Both of the cDNA fragment have been cloned in E.coli.  Ringkasan Karat daun yang disebabkan oleh jamur Hemileia vastatrix merupakan salah satu penyakit penting pada perkebunan kopi arabika. Untuk memahami mekanisme ketahanan kopi arabika terhadap karat daun, penelitian ini bertujuan untuk mempelajari ekspresi gen β-1,3 glukanase dan kitinase pada varietas kopi arabika S1934 yang dilaporkan tahan karat daun dan varietas BLP10 yang termasuk rentan karat daun. Untuk itu kedua varietas diuji kembali ketahanannya terhadap H. vastatrix melalui bioesai dan dilakukan RT-PCR menggunakan RNA total dari S1934 dan BLP10, baik yang diinokulasi dengan H. vastatrix maupun yang tidak diinokulasi, untuk mempelajari ekspresi gen GLU dan CHI. Dua pasang primer spesifik dirancang untuk mengamplifikasi daerah konservatif kedua gen  tersebut. Hasil amplifikasi disekuen dan dianalisis menggunakan program BlastX. Hasil bioesai mengkonfirmasi bahwa S1934 tahan terhadap H. vastatrix, sedangkan  BLP10 rentan.  β-1,3 glukanase diekspresikan pada kedua varietas, baik yang diinokulasi maupun yang tidak diinokulasi, namun dengan tingkat ekspresi yang sedikit berbeda. Varietas S1934 mengekspresikan β-1,3 glukanase lebih tinggi dibandingkan dengan BLP10. Ekspresi gen tersebut pada S1934 yang diinokulasi lebih tinggi dibandingkan dengan yang tidak diinokulasi. Sedangkan kitinase hanya diekspresikan pada varietas S1934. Hasil sekuensing dan analisis DNA mengkonfirmasi bahwa sekuen hasil RT-PCR merupakan bagian ekson dari gen penyandi β-1,3 glukanase dan kitinase. Kedua fragmen tersebut telah diklon pada E. coli.
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
Menara Perkebunan Vol. 68 No. 2: 68 (2), 2000
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v68i2.138

Abstract

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. 
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
Menara Perkebunan Vol. 68 No. 2: 68 (2), 2000
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v68i2.139

Abstract

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. 
Solubilization of insoluble phosphates by Aspergillus niger Pelarutan fosfat sukar larut oleh Aspergillus niger LAKSMITA P SANTI; D H GOENADI; . SISWANTO; I SAILAH; . ISROI
Menara Perkebunan Vol. 68 No. 2: 68 (2), 2000
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v68i2.142

Abstract

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.
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
Menara Perkebunan Vol. 75 No. 2: 75 (2), 2007
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v75i2.146

Abstract

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.
Transformasi kopi robusta (Coffea canephora) dengan gen kitinase melalui Agrobagterium tumefaciens LBA4404 Transformation of robusta coffee (Coffea canephora) with chitinase gene mediated by Agrobacterium tumefaciens LBA4404 . SISWANTO; Fetrina OKTAVIA; Asmini BUDIANI; . , SUDARSONO; . PRIYONO; Surip MAWARDI
Menara Perkebunan Vol. 71 No. 2: 71 (2), 2003
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v71i2.162

Abstract

SummaryGenetic engineering of robusta coffee forresistance to pathogenic fungi is considered to beone of the potential approaches to overcome theproblem at robusta coffee plantation caused bypathogenic fungi. This research was aimed tointroduce chitinase (CHI) gene into embryogeniccalli of robusta coffee and regenerate theplantlets. Embryogenic calli were co-cultivatedwith Agrobacterium tumefaciens LBA4404harboring pCAMBIA1301 which containschitinase gene under 35S promoter. In thisresearch four concentrations (0, 50, 100 and150 mg/L) of acetosyringone (AC) were used inthe co-cultivation medium. Selection fortransformed calli was conducted by graduallyincreasing the concentration of hygromicin from5 to 25 mg/L. Somatic embryo (SE) was inducedfrom callus on the medium containing acombination of BAP 5 mg/L and IAA (0, 0.25 or0.50 mg/L). Integration CHI in plant genome wasexamined by GUS assay and PCR. The resultrevealed that among the four AC concentrationstested, 100 mg/L gave the highest percentage ofcalli growing on the selection medium (42.5%).BAP concentration of 5 mg/L alone was the mosteffective for inducing of SE from transformedcalli with the highest percentage of 43.1% andaverage number SE of 8.8 ± 3. The strongestGUS expression on the calli at 3 days aftertransformation and the calli grown on selectionmedium containing 150 mg/L AC, which were56.5% and 40% respectivelly. PCR analysisshowed that 7 out of 12 plantlets tested,contained CHI gene. From this research 28transgenic plantlets of robusta coffee wereobtainedRingkasanRekayasa genetika untuk merakit tanamankopi robusta tahan jamur pathogen dipandangmerupakan salah satu pendekatan alternatif yangpotensial untuk mengatasi masalah padaperkebunan kopi robusta akibat serangan jamurpatogen. Penelitian ini bertujuan untuk meng-introduksikan gen kitinase (CHI) ke dalam kalusembriogenik kopi robusta dan regenerasinyamenjadi planlet, sebagai upaya untuk merakittanaman kopi robusta tahan serangan jamur.Kalus embriogenik diko-kultivasi denganAgrobacterium tumefaciens LBA4404 pembawapCAMBIA1301 yang mengandung gen kitinasedi bawah kontrol promotor 35S. Pada percobaanini, empat konsentrasi asetosiringon (AS) (0, 50,100 dan 150 mg/L) digunakan dalam medium ko-kultivasi. Seleksi kalus hasil transformasidilakukan dengan peningkatan konsentrasi higro-misin secara bertahap dari 5 mg/L sampai25 mg/L. ES diinduksi dari kalus pada mediumyang mengandung BAP 5 mg/L dan IAA (0; 0,25dan 0,50 mg/L). Integrasi gen CHI ke dalamgenom tanaman dianalisis melalui uji GUS danPCR. Hasil penelitian menunjukkan bahwa darikeempat konsentrasi AS yang diuji, AS 100 mg/Lternyata menghasilkan persentase tertinggi kalusyang tumbuh pada medium seleksi (42,5%).Konsentrasi BAP 5 mg/L tanpa penambahan IAAefektif menginduksi ES dari kalus hasiltransformasi dengan persentase tertinggi 43,1%dan rata-rata jumlah ES 8,8±3. Ekspresi GUStertinggi dideteksi pada kalus tiga hari setelahtransformasi dan kalus yang tumbuh di mediumseleksi yang mengandung AS 150 mg/L,masing-masing 56,5% dan 40,0 %. Analisis PCRmenunjukkan bahwa 7 planlet dari 12 planletyang diuji, membawa gen CHI. Dari penelitianini dihasilkan 28 planlet kopi robusta transgenik.
Produksi dan karakterisasi lakase Omphalina sp. Production and characterization of Omphalina sp. laccase . SISWANTO; . SUHARYANTO; Rossy FITRIA
Menara Perkebunan Vol. 75 No. 2: 75 (2), 2007
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v75i2.177

Abstract

SummaryOmphalina sp. a white-rot fungi (WRF)originated from oil palm plantation has abilityto degrade empty fruit bunches of oil palm(EFBOP) so that it is expected to producelaccase with high activity. The ability ofOmphalina sp. to produce laccase enzyme onliquid fermentation will be studied. The enzymewill also be partially purified andcharacterized. The research result showed thatthe highest enzyme activity (1.162 U/mL) wasobtained using glucose malt yeast (GMY)medium at room temperature for four days.The addition of 2,5-xylidine as an inducerproduced laccase earlier i.e two days, but theactivity of laccase was less active afterprolonged incubation compared to that ofcontrol. The laccase produced on mediumcontaining 2% EFBOP reached optimumactivity as much as 0.38 U/mL after 10 th daysof incubation. Partial purification of laccaseon Sephacryl S-200 HR column resulted58.23% of yield recovery with twice purity thanbefore. The optimum pH of laccase was 4.5.Laccase activity was stable even after heatedon 50 o C for 30 minutes, but then decreasedwhen heated until 60 o C. The laccase has K Mand V max as much as 0.15 mM and 0.56 U/mLrespectively.RingkasanOmphalina sp., adalah fungi pelapuk putih(FPP) hasil isolasi dari kebun kelapa sawityang diketahui mampu mendegradasi tandankosong kelapa sawit (TKKS) dengan cepatsehingga diharapkan mampu menghasilkanlakase dengan aktivitas tinggi. KemampuanOmphalina sp. menghasilkan enzim lakasepada fermentasi cair akan dipelajari. Selain itu,lakase yang dihasilkan akan dimurnikan secaraparsial serta dilakukan karakterisasi pH, suhu,dan konsentrasi substrat optimum. Hasilpenelitian menunjukkan bahwa Omphalina sp.menghasilkan lakase dengan aktivitas tertinggi(1,162 U/mL) pada medium glucose malt yeast(GMY) yang diinkubasikan pada suhu ruangselama empat hari. Penambahan 2,5-xilidinsebagai induser mempercepat produksi lakaselebih awal yaitu dalam waktu dua hari, namunaktivitasnya masih lebih rendah dibandingkandengan kontrol pada inkubasi lebih lanjut.Lakase dari Omphalina sp. juga dapatdiproduksi pada medium yang mengandung2% TKKS dan aktivitasnya mencapai0,38 U/mL yang diinkubasi dalam suhu ruangselama 10 hari. Pemurnian parsial pada kolomSephacryl S-200 HR menghasilkan rendemensebesar 58,23% dengan kemurnian dua kalinya.Aktivitas lakase optimum pada pH 4,5 dantetap stabil setelah pemanasan selama 30 menitpada suhu ruang hingga 50 o C dan menuruntajam pada suhu 60 o C. Lakase Omphalina sp.menghasilkan nilai K M dan V maks masing-masing sebesar 0,15 mM dan 0,56 U/mL.
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
Menara Perkebunan Vol. 84 No. 1 (2016): 84 (1), 2016
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v84i1.213

Abstract

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.
Co-Authors . , SUDARSONO . ISROI . PRIYONO A BUDIANI Alia Damayanti Asmini Budiani D A SANTOSO D H GOENADI D SANTOSO Deden Dewantara ERIS Fetrina Oktavia H MINARSIH Hanny Hafiar HAPPY WIDIASTUTI Haryo Tejo PRAKOSO I Gusti Bagus Wiksuana I SAILAH I SUSANTI Imron RIYADI Irma KRESNAWATY Irma Kresnawaty LAKSMITA P SANTI Maemonah, Maemonah Rossy FITRIA S DWI UTOMO S MAWARDI Sri Wahyuni Suharyanto Sumi Hudiyono Surip Mawardi T CHAIDAMSARI Tri Panji Yoharmus SYAMSU