MOLECULAR CHARACTERIZATION OF A GREEN ALGAE ISOLATE BY 16S rRNA IN IMPROVEMENT OF CAROTENOID PRODUCTION

Sintesis karotenoid alami belum pernah melebihi produk sintetik pada skala komersial. Kurangnya pemahaman mengenai aspek mikrobiologis dan ekofisiologis isolat penghasil karotenoid seringkali menyebabkan terjadinya kesalahan penamaan spesies. Sam isolat lokal alga hijau dan Perairan Jepara yang digunakan sebagai pakan alami sumber karotenoid hewan-hewan perikanan, pada mulanya dianggap sebagai Dunaliella. Penamaan Dunaliella hanya dilakukan berdasarkan pengamatan mikrobiologis dan ekofisiologis yang kurang lengkap. Tujuan utama penelitian mi adalah menentukan spesies sam isolat lokal alga hijau secara molekuler menggunakan 16S rDNA untuk mendeteksi jalur biosintesis karotenoid yang digunakan. Urutan basa 16SrRNA yang diperoleh dianalisis menggunakan Multiple Alignment Analysis dan analisis filogenetik melalui program ClustalX ClustaiW, GeneDoc, Phylip dan NjPlot. Hasil penelitian memperlihatkan bahwa Isolat alga hijau menunjukkan similaritas yang tinggi dengan anggota-anggota Sianobakteria. Keserupaan tertinggi dimiliki dengan Cyanobacterium sp. MBIC 1021 sebesar 99 %, diikuti Synechoystis PCC6308 sebesar 95 %. Hasil analisis similaritas dan filogenetik memperlihatkan peluang bahwa Isolat alga hijau mengikuti jalur baru non-mevalonat dalam biosintesis karotenoidnya. Abstract Carotenoids production levels are not yet competitive with carotenoid levels presently produced by fermentation, synthesis and isolation. An attempt to optimize carotenoid production from local isolate of green algae from BBAP Jepara has faced several problems, primarily related to the microbiological and eco-physiological characteristic which affecting growth that have not sufficiently been understood. A misnamed of species also have arisen due to wrong characterization. One local isolate of Dunaliella species from BBAP Jepara was found potentially useful as source of carotenoids in food additives or as food supplement in fish farming. The present study aimed to characterize the species of green algae isolate from Jepara waters based on molecular techniques using 16S rRNA approach to detect its carotenoid biosynthetic pathway. Similarities analysis and phylogenetic relationship of 16S rRNA sequence was analyzing with Multiple Alignment Analysis by ClustalX ClustalW, GeneDoc, Phylip and NjPlot Programs. Molecular analysis showed close relationship among isolate of green algae and Cyanobacteria with 99 % similarity with Cyanobacterium sp. MBIC 1021 and 95 % similarity with Synechocysti.s PCC3O8. The result of this analysis indicated possibilities that a green algae isolate following the new non-mevalonate pathway for its catotenoid biosynthetics. Keywords A green algae isolate, Dunaliella, 16S rRNA, Cyanobacteri

Energi Mandiri dengan Pemanfaatan Limbah Cair pada Industri Pabrik Kelapa Sawit

Limbah merupakan permasalahan yang cukup serius di dunia industri kelapa sawit dan berdampak merusak lingkungan. Sehingga ditemukan teknologi biogas untuk mengatasi limbah cair (POME) dan dimanfaatkan untuk menghasilkan listrik. Teknologi tank reaktor cukup efektif dalam penanganan limbah cair (POME) untuk menghindari gas metan terlepas ke udara atmosfir. Pengembangan teknologi biogas di Indonesia terus dilakukan dari pendekatan skala besar dan kecil. Dari kapasitas produksi pabrik sebesar 30 ton/jam Tandan Buah Segar (TBS) dapat menghasilkan tenaga listrik sebesar  1 MW dari PLT Biogas (PLTBg)

Isolasi Bakteri Endofit Tanaman Pepaya (Carica Papaya L.) Dan Uji Aktivitas Enzim Amilase

ABSTRACTEndophytic bacteria are beneficial microorganisms that interact with plants without causing disturbance or damage to them. Research shows that certain endophytic bacterial isolates can produce secondary metabolites that have health effects, especially endophytic bacteria isolated from the Papaya plant (Carica Papaya. L). Amylolytic bacteria is one of the bacteria that has the potential to produce amylase enzymes which play an important role in industry. Endophytic bacteria were isolated using Nutrient Agar media 1% starch (soluble starch) and amylolytic index measurements were carried out using Lugol's reagent. Isolation of endophytic bacteria was carried out by streak method on Nutrient Agar media. The cultures were incubated at 37ºC for 48 hours. The morphological characteristics of the growing colonies were observed and Gram stained was carried out. Amylase activity was measured using the Dinitrosalisilic acid (DNS) method. The results of this study found 19 isolates of endophytic bacteria were isolated. There were 5 isolates that had the highest clear zone, namely isolates AE4, BE2, DE6, TD2, and TD5. The highest amylase enzyme activity of 0.225 U / mL was produced in TD5 isolates.Keyword : Isolation, Endophytic Bacteria, Enzyme Amylase, Papaya Plant

Improvement of astaxanthin production from Phaffia rhodozyma by protoplast fusion

Protoplast fusion between intrastrain of P. rhodozyma has been generated in order to increase astaxanthin production. The best protoplast was obtained under osmotic stabilizer using buffer phosphate with the addition of 0.1 M CaC12. Higher frequency of fusion and regeneration were obtained under 8 mg /ml lysozyme and 35 °/0 PEG 4000 treatment. Frequency of regeneration was 36 - 97%, frequency of fusion was 86.5 %. Several hybrids showed two-fold increase levels of ploidy, while yields of carotenoid pigments increased up to 2.03 times than the levels of wild type. Carotenoids accumulated to a level as high as 2. 529 lig /g dcw with the estimation of astaxanthin production reached the value of 488.75 lig/g dcw. Keywords: astaxanthin â Phaffia rhodozyma âprotoplast fusio

Exploiting Tofu Liquid Waste Become Healthy Food by Lactic Acid Bacteria and Determination of Its Cholesterol Levels in Vitro

One of the way to overcome problem contamination of Tofu Liquid Waste (TLW) is by exploiting again the waste so that can yield high economic valuable product. The aim of this study is to make functional beverage from Tofu liquid waste with Lactic Acid Bacteria (LAB) . The aim of the research was to make a healthy food (yoghurt) from combination of TLW and fresh milk that fermented by Lactic Acid Bacteria, such as Lactobacillus bulgaricus Streptococcus thermophillus and Lactobacillus bulgaricus and study its activity in degrading cholesterol levels. At the research was conducted by making yoghurt with 5 treatments of TLW concentration (0, 25, 50,75, and 100 ). Result of this research indicate that yoghurt with various of TLW concentration can degrade cholesterol in vitro equal to 63,33 %, so that yoghurt can be made as functional food, Yoghurt with 25% TLW had the good Yoghurt criteria and its organoleptic value not different with control ( %TLW

KARAKTERISTIK DAN SIFAT KINETIKA ENZIM KITINASE ASAL JAMUR ENTOMOPATOGEN Beauveria bassiana

Characteristics and Kinetics of Chitinase Enzyme from Entomopathogenic Fungus Beauveria bassianaBeauveria bassiana is one of the entomopathogenic fungi that produces chitinase when infecting the host. Chitinase is widely used as biocontrol agents because it can degrade chitin into an environmentally friendly product. This study aims to characterize and test the kinetics of chitinase from B. bassiana. This characterization includes determination of pH and optimum temperature, enzyme stability and enzyme kinetics test by determining Km and Vmax value with Lineweaver-Burk equations. The result of experiment showed that the chitinase B. bassiana had pH and optimum temperature of 5 and 40ºC respectively. This enzyme was stable until 90 minutes incubation at 40ºC. The Km and Vmax values were 0.181 mg/L and 0.022 mg/L.sec respectively. The Km value is higher than Vmax, which means the affinity of the enzyme to the lower substrate requiring high substrate concentration to increase the reaction rate. It can be concluded that the chitinase activity of B. bassiana is still low.Keywords: Beauveria bassiana, characteristics and kinetics, chitinase enzyme, entomopathogenic, Lineweaver-BurkABSTRAKBeauveria bassiana merupakan salah satu jamur entomopatogen yang memproduksi kitinase saat menginfeksi inangnya. Enzim kitinase saat ini banyak digunakan sebagai agen biokontrol karena dapat mendegradasi kitin menjadi produk yang ramah lingkungan. Penelitian ini bertujuan untuk mengkarakterisasi dan menguji kinetika enzim kitinase asal jamur B. bassiana. Metode yang digunakan dalam karakterisasi ini mencakup penentuan pH dan suhu optimum, kestabilan enzim pada suhu optimumnya, dan uji kinetika enzim yang mencakup penentuan nilai Km dan Vmaks dengan persamaan Lineweaver-Burk. Hasil penelitian karakterisasi menunjukkan bahwa enzim kitinase B. bassiana mempunyai pH dan suhu optimum masing-masing 5 dan 40ºC. Enzim ini stabil sampai pada 90 menit inkubasi pada suhu 40ºC. Nilai Km diperoleh 0,181 mg/L dan Vmaks sebesar 0,022 mg/L.detik. Nilai Km lebih tinggi daripada Vmaks, yang artinya afinitas enzim terhadap substrat rendah sehingga membutuhkan konsentrasi substrat yang tinggi untuk meningkatkan kecepatan reaksi, maka dapat disimpulkan bahwa aktivitas kitinase dari B. bassiana masih tergolong rendah.Kata kunci: Beauveria bassiana, entomopatogen, enzim kitinase, karakteristik dan kinetik, Lineweaver-Bur

ISOLASI KHAMIR DARI BATANG TANAMAN TEBU DAN IDENTIFIKASINYA BERDASARKAN SEKUENS INTERNAL TRANSCRIBED SPACER

Isolation of Yeasts from Sugarcane Stems and Their Identification Based on Internal Transcribed Spacer Sequences ABSTRACTFermentative yeasts used in food, health, and energy industries need to be explored to discover their potential. The purpose of this study was to obtain fermentative yeast isolates from sugarcane stems and subsequently to undertake morphological, biochemical, and molecular identification. The isolation of epiphytic and endophytic yeasts was carried out by spread plate method using sugarcane soak water and sugarcane juice on potato dextrose agar (PDA) and yeast-glucose-peptone (YGP) agar media. Morphological identification was based on macroscopic and microscopic observations. Biochemical identification was performed using carbohydrate fermentation and 50%-glucose media tests. Selected isolates were identified molecularly using Internal Transcribed Spacer (ITS). Seven yeast isolates were obtained, of which isolate Ed 1B was selected. Isolate ED 1B was of round colonies, creamy white colour, shiny, embossed, and wavy appearance, ovoid cell shape with a cell diameter of 4.74 µm. It had budding cells, was able to ferment glucose and sucrose (but not lactose), and grew on 50 %-glucose media. Results of BLAST showed that isolates Ed 1B had 99% homology with Kodamaea ohmeri.Keywords: isolation, ITS, molecular identification, Saccharum officinarum L., yeast ABSTRAKKhamir fermentatif yang digunakan dalam industri pangan, kesehatan dan energi perlu dieksplorasi untuk mengetahui potensinya. Tujuan penelitian ini adalah untuk memperoleh isolat khamir fermentatif dari batang tebu dan untuk kemudian diidentifikasi secara morfologi, biokimia dan molekuler. Isolasi khamir epifit dan endofit dilakukan dengan metode cawan sebar dari air rendaman tebu dan jus tebu pada media potato dextrose agar (PDA) dan yeast-glucose-peptone (YGP). Identifikasi morfologi berdasarkan pengamatan makroskopis dan mikroskopis. Identifikasi biokimia menggunakan uji fermentasi karbohidrat dan uji media glukosa 50%. Isolat terpilih diidentifikasi molekuler menggunakan Internal Transcribed Spacer (ITS). Hasil isolasi memperoleh 7 isolat khamir. Satu isolat terpilih (Ed 1B) didapatkan dan memiliki ciri-ciri koloni bulat, putih krem, mengkilap, timbul, bergelombang, bentuk sel ovoid dengan diameter sel 4,74 µm, memiliki budding cell, mampu memfermentasi glukosa dan sukrosa, tidak memfermentasi laktosa, serta tumbuh pada media glukosa 50%. Hasil BLAST menunjukkan bahwa isolat Ed 1B memiliki homologi 99% dengan Kodamaea ohmeri.Kata Kunci: identifikasi molekuler, isolasi, ITS, khamir, Saccharum officinarum L

ISOLASI DAN KARAKTERISASI BAKTERI ASAM LAKTAT DARI PANGAN FERMENTASI CINCALOK SEBAGAI PENGHASIL GAMMA-AMINOBUTYRIC ACID

Isolation and Characterization of Lactic Acid Bacteria from Fermented Food Cincalok as Producer of Gamma-Aminobutyric Acid Cincalok is a fermented food originating from West Kalimantan. This study aimed to obtain lactic acid bacterial isolates (LAB) capable of producing gamma-aminobutyric acid (GABA), to characterize the LAB isolates obtained, and to obtain GABA by the Thin Layer Chromatography (TLC) method. Bacterial growth and GABA production was carried out by adding 5% MSG and without MSG, and measured spectrophotometrically. In this study, 4 LAB bacterial isolates were obtained which were coded CIN-1, CIN-2, CIN-3, and CIN-4. GABA identification of all the LAB isolates using TLC Silica Gel 60 F254 with butanol: acetic acid: distilled water (5: 3: 2) as eluent yielded Rf 0.61 and Rf MSG 0.38. The highest growth was achieved by isolate CIN-3 with an absorbance of 1.488 (at 48 hour) in non-MSG medium, while the addition of 5% MSG resulted in an absorbance of 1.631 (at 42 hour). GABA production was achieved by isolate CIN-3 with 5% MSG treatment with a concentration of 201.472 mM and without MSG with a concentration of 171.195 mM. Cincalok merupakan pangan fermentasi yang berasal dari Kalimantan Barat. Penelitian ini bertujuan untuk mendapatkan isolat bakteri asam laktat (BAL) yang mampu menghasilkan gamma-aminobutyric acid (GABA), melakukan karakterisasi isolat BAL yang diperoleh dan dapat diperoleh GABA dengan metode Kromatografi Lapis Tipis (KLT). Penumbuhan bakteri dan produksi GABA dilakukan dengan penambahan MSG 5% dan tanpa MSG, dan diukur menggunakan spektrofotometer. Dalam penelitian ini diperoleh 4 isolat bakteri BAL yang diberi kode CIN-1, CIN-2, CIN-3, dan CIN-4. Identifikasi GABA dari semua isolat BAL tersebut menggunakan KLT Silica Gel 60 F254 dengan eluen butanol: asam asetat: aquades (5: 3: 2), menghasilkan Rf 0,61 dan Rf MSG 0,38. Pertumbuhan tertinggi terjadi pada isolat CIN-3 non MSG dengan absorbansi 1,488 (jam ke-48), sedangkan dengan penambahan MSG 5% menghasilkan absorbansi 1,631 (jam ke-42). Produksi GABA dicapai isolat CIN-3 dengan perlakuan MSG 5% dengan konsentrasi 201.472 mM dan tanpa MSG dengan konsentrasi 171,195 mM

Efektivitas Limbah Kulit Kering Nanas Madu (Ananas Comosus l.Merr) untuk Pembuatan Bioetanol dengan Proses Fermentasi dan Distilasi

Bioenergi adalah salah satu energi yang dapat diperbaruhi dan juga ramah lingkungan, salah satunya adalah bioetanol. Bahan baku yang mengandung glukosa, selulosa dan pati dapat digunakan menjadi bioetanol Nanas madu adalah tanaman yang dapat dimanfaatkan menjadi bioenergi. Penelitian ini bertujuan membuktikan efektivitas limbah kulit nanas madu kering untuk pembuatan bioetanol serta mengetahui kadar bioetanol yang dihasilkan. Proses fermentasi yang digunakan dalam peneltian ini . Kulit nanas madu yang sudah dikeringkan di proses hidrolisis, fermentasi dan distilasi. Proses fermentasi dengan variasi waktu fermentasi 1 hari, 2 hari, 3 hari, 4 hari, 5 hari, 6 hari, 7 hari,8 hari, 9 hari, 10 hari, 11 hari dan 12 hari. Hasil fermentasi tercepat akan didistilasi sebanyak dua kali dan dianalisa kadar bioetanol dengan GC-MS. Kadar bioetanol dari limbah kulit kering nanas madu didapatkan sebesar 95,66 % dengan waktu fermentasi selama 4 hari

Pengaruh Plastik Pengemas Low Density Polyethylene (Ldpe), High Density Polyethylene (Hdpe)dan Polipropilen (Pp)terhadap Penundaan Kematangan Buah Tomat (Lycopersicon Esculentum.mill )

The increasing demand of tomatoes corresponds to the economic and population growth. To maintain the quality of tomatoes, an effort to holdup the tomato ripeness was done using several kinds of plastic packaging. This research was intended to assess the effects of using plastic packaging and the most effective kind of plastic packaging to extend tomato storage period. The research was done at BSFT Biology Laboratory Diponegoro University and Food Technology Laboratory Soegijapranata Catholic University. The design used in the research was Completely Randomized Design (CRD) with some treatments using different plastic packaging type (P 0 = control / without packaging, P1 = Low Density Polyethylene (LDPE), P2 = High Density Polyethylene (HDPE) and P3 = Polypropylene (PP), each treatment was repeated three times. The parameter observed was the percentage of weight loss, the change in color (using chromameter method); the hardness level (texture analyzer method) and total content of sugar. Data were analyzed by Analysis of Variance (ANOVA) then continued with Duncan's Multiple Range Test (DMRT) at the 95% significance level to find out the difference. The results show that the use of plastic as packaging materials is able to delay tomato ripeness and the effective type plastic-based packaging to weight loss and change color is HDPE and PP