Judul: Metode K-Rataan Terpangkas dalam Mengatasi Pencilan pada Data Bergerombol
Abstrak: Metode penggerombolan merupakan salah satu alat yang sering digunakan dalam analisis data. Pada saat pembentukan gerombol, terkadang ada beberapa data yang terletak jauh dari gerombolnya, yang disebut sebagai pencilan. Adanya pencilan dapat mengakibatkan bergesernya titik pusat dari sebuah gerombol. Keadaan ini menunjukkan perlunya merancang prosedur penggerombolan baru dengan penekanan pada sifat kekekaran. Salah satu prosedur yang dapat digunakan adalah metode k-rataan terpangkas (trimmed k-means). Penelitian ini bertujuan untuk membandingkan ketepatan klasifikasi hasil penggerombolan dari metode k-rataan dengan metode k-rataan terpangkas pada beberapa kondisi keragaman data. Berdasarkan kajian yang telah dilakukan melalui pembangkitan data, metode k-rataan terpangkas dapat menentukan anggota gerombol dengan ketepatan yang lebih tinggi dibandingkan dengan metode k-rataan.
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Judul: Deteksi Gerombol Dengan Metode k-Rataan Kernel Gauss
Abstrak: Metode penggerombolan k-rataan tidak mampu menggerombolkan data yang terpisah secara non linier. Salah satu cara untuk menanggulangi permasalahan ini adalah dengan menggunakan fungsi kernel. Penerapan metode k-rataan di dalam ruang transformasi fungsi kernel dikenal dengan metode k-rataan kernel. Tujuan dari penelitian ini adalah untuk mengkaji nilai lebar jendela dan persentase salah klasifikasi metode k-rataan kernel pada beberapa jenis gerombol data. Data yang digunakan dalam penelitian ini terdiri dari data simulasi dan data asli. Hasil penelitian menunjukkan bahwa metode k-rataan kernel mampu menggerombolkan gerombol-gerombol yang terpisah secara linier maupun non linier sedangkan metode k-rataan hanya mampu menggerombolkan gerombol-gerombol yang terpisah secara linier. Pada data asli, metode k-rataan kernel menghasilkan persentase kesalahan klasifikasi yang lebih kecil dibandingkan metode krataan. Kedua metode memiliki kelemahan dalam menggerombolkan gerombol-gerombol yang memiliki anggota tumpang tindih. Penentuan lebar jendela pada fungsi kernel Gaussian sangat berpengaruh terhadap persentase salah klasifikasi. Penentuan lebar jendela dengan perkiraan kasar cukup efisien.
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Judul: Extracellular Expression of thermostable Geobacillus stearothermophilus T1.2RQ Lipase for Biodiesel Production
Abstrak: The extracellular expression of enzymes in an excellent secretion host such as Bacillus subtilis is a useful strategy for making enzymes of industrial significance more affordable by lowering the cost of its downstream processing. Lipases are a category of hydrolases that catalyze the hydrolysis of triglycerides to glycerol and free fatty acids. Also, lipases catalyze the hydrolysis and transesterification of other esters as well as the synthesis of esters. They can perform very specific biotransformation reactions and are widely used in food, organic synthesis, detergents, cosmetic, and pharmaceutical industries. Geobacillus stearothermophilus T1.2RQ lipase is an industrially-desired thermostable lipolytic enzyme with an excellent hydrolytic and transesterification activity. This study aimed to express T1.2RQ lipase, in B. subtilis WB800 and ensure its secretion into the extracellular medium. Three signal peptides (AmyQ, Epr, LipA) and two promoters (Pgrac01 and Pgrac100) were used in the secretory-expression of T1.2RQ lipase, based on five constructed expression vectors constructed by restriction cloning and PCR mutagenesis. Sangers sequencing was used to verify constructs to ensure they are in the correct reading frame. Recombinants vectors, pHT43 (plasmid control), pHT43-T1.2RQ, pHT43- T1.2RQ_AmyQ, pHT43-T1.2RQ_Epr, and pHT43-T1.2RQ_LipA, were transformed by electroporation into Bacillus subtilis WB800. The electroporation was successful with the optimum condition of 2500V, 200 ohms, 25µF, and 5milliseconds. Recombinant B. subtilis pHT43 (negative control) and pHT43-T1.2RQlip + signal peptides were streaked on the lipidic agar, LA+TBN. All four recombinants, including the negative control (B. subtilis WB800-pHT43), produced clear zones on the TBN agar plate after overnight incubation (fig 2), indicating the presence of a lipolytic enzyme in all of the recombinant B. subtilis cells. pHT43-T1.2RQlip + signal peptides (AmyQ, Epr, and LipA) have clear zones with significantly higher diameters than B. subtilis WB800-pHT43 (negative control). This phenomenon suggests that the recombinant thermostable lipase T1.2RQ introduced into these cells was successfully expressed, thereby increasing the lipolytic capability of its host cells T1.2RQ lipase was expressed in 50ml terrific broth medium (TB) and the expression induced with 1mM IPTG. Lipase activity assay using p-nitrophenyl esters showed that all three signal peptides directed the secretion of T1.2RQ lipase into the extracellular medium. Signal peptide, LipA, resulted in the highest extracellular activity of 5.6 U/mL, which corresponds to a 6-fold increase over the parent B. subtilis WB800 strain. Meanwhile, signal peptides, AmyQ and Epr produced 4.2 U/mL and 3.4 U/mL activity respectively. SDS-Zymogram analysis confirmed that T1.2RQ lipase was correctly processed and secreted in its original size of 44 kDa. An attempt to further increase lipase expression level by promoter optimization showed that, contrary to expectation, promoter Pgrac100 exhibited similar expression levels as the original Pgrac01. Nevertheless, the demonstration of the successful secretion of T1.2RQ lipase in this work confirms the latter as a promising approach for the industrial production of the enzyme
Keyword: Bacillus subtilis, Expression, Extracellular, Heterologous, Lipase, Thermostable