Judul: Pendugaan parameter model dinamik dengan menggunakan metode Robust dan implementasinya ke dalam Add-ons mathematica
Abstrak: Salah satu tahapan dalam pemodelan matematika adalah pendugaan parameter model yang telah ditetapkan. Metode yang umum digunakan dalam pendugaan parameter model tersebut adalah metode kuadrat terkecil (least square). Namun demikian, metode tersebut telah diketahui rentan terhadap pengaruh data pencilan (outliers). Oleh karena itudiperlukan metode lain yang bersifat robust atau tahan terhadap pengaruh pencilan. Metode robust yang dimaksud antara lain: metode kuadrat terkecil terboboti (weighted least square), metode simpangan mutlak terkecil (least absolute value), dan metode kuadrat terkecil terpangkas (least trimmed square).
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Judul: Pendugaan parameter model dinamik dengan metode robust kuadrat terkecil tertimbang dan simpangan mutlak
Abstrak: Misalkan diketahui segugus data yang mengandung pencilan dan representasi model dinamiknya maka secara umum hasil pendugaan parameter model tersebut dengan menggunakan metode kuadrat terkecil kurang memuaskan. Hal tersebut karena metode kuadrat terkecil sangat sensitif dengan keberadaan pencilan. Jadi perlu dicari metode lain yang lebih tahan dengan keberadaan pencilan. Metode tersebut adalah metode robust. Ada beberapa jenis metode robust antara lain metode kuadrat terkecil tertimbang dan metode simpangan mutlak. Setiap amatan pada metode kuadrat terkecil diberi penimbang yang sama. Sedangkan pada metode kuadrat terkecil tertimbang dan simpangan mutlak, setiap amatan diberi penimbang sesuai dengan besar sisaannya. Semakin besar sisaannya maka penimbang yang diberikan akan semakin kecil. Karena itulah metode kuadrat terkecil tertimbang dan simpangan mutlak lebih tahan terhadap pencilan. Pendugaan parameter metode kuadrat terkecil, kuadrat terkecil tertimbang dan simpangan mutlak dari kedua model dalam tulisan ini diimplementasikan ke dalam pemprograman fungsional berbasis sistem aljabar komputer seperti Mathematica. Model yang digunakan untuk mengkaji ket iga metode pendugaan parameter tersebut adalah model logistik dan model SEI. Hasil yang didapat adalah grafik dengan parameter dugaan metode robust dari kedua model akan mengikuti tebaran data secara umum. Selain itu sisaan relatif yang didapat menggunakan metode robust juga lebih terkonsentrasi dibandingkan metode kuadrat terkecil. Hal tersebut memperlihatkan bahwa metode robust lebih tahan terhadap pencilan dibandingkan dengan metode kuadrat terkecil.
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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