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"Protein apoptin dari virus anemia ayam telah diteliti memiliki potensi yang baik sebagai pendeteksi dini sel kanker. Keberhasilan produksi apoptin yang tidak lagi berupa badan inklusi telah memberikan harapan lebih besar untuk melakukan optimasi produksi protein apoptin rekombinan ini. Sel rekombinan apoptin yang berhasil diproduksi dalam skala besar dengan menggunakan inang Bacillus subtilis 168 pOXGW-apop-2His8Arg, pOGW-apop-12His dalam berbagai variasi kondisi kultivasi (konsentrasi substrat penginduksi, laju aerasi, dan laju agitasi) kemudian dipurifikasi menggunakan metode IMAC dalam kolom afinitas (HisTrap FF 5 mL) yang berisi ion logam transisi Ni2+ dengan menggunakan instrumen AKTA Prime Plus. Secara rata-rata, hasil purifikasi menunjukkan bahwa elusi apoptin rekombinan terjadi ketika nilai konduktivitas berada pada angka 15,85 mS/cm, dengan konsentrasi imidazole berada pada kisaran nilai 76% - 88% (384,8-442,4 mM), yang terlihat dari grafik gradien elusi. Pengukuran konsentrasi protein apoptin dengan menggunakan metode Bradford menujukkan bahwa konsentrasi terbesar diperoleh pada sampel dengan sistem agitasi 250 rpm dan laju aerasi 0,5 Nl/menit, dengan besar konsentrasi 0,0507 mg/ml. Hasil purifikasi berhasil dideteksi menggunakan SDS-PAGE 12% dengan hasil pita protein terlihat di area 15 kDa dan 58,5 kDa untuk semua sampel.

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Apoptin has been known to be having a great potency for cancer detection. The success of apoptin production which is not in inclusion body form anymore has given a bigger hope to optimize its production. Recombinant apoptin cells which was succesful to be cultivated in large scale using Bacillus subtilis 168 pOXGW-apop-12His8Arg, pOGW-apop-12His vector in various cultivation condition (aeration rate, agitation rate) then purified using IMAC method in Ni2+-loaded affinity column (HisTrap FF 5ml) and proceeded in AKTA Prime Plus instrument. Averagely, purifcation result showed that the elution of apoptin recombinant protein happened when the conductivity value at 15,85 mS/cm, with imidazole concentration lied around 76%-88% (384,8-442,4 mM), which could be seen from elusion gradient curve. The measurement of apoptin protein concentration using Bradford method showed that the biggest concentration was obtained from the sample with agitation rate 250 rpm and aeration rate 0,5 Nl/min, and the value is 0,0507 mg/ml. Purification yield was succesfully detected using SDS-PAGE 12%, with protein band was seen on 15 kDa and 58,5 kDa area for all samples."

"DNA Polimerase merupakan enzim sintesis DNA yang dimanfaatkan dalam metode amplifikasi DNA Polymerase Chain Reaction (PCR) untuk deteksi penyakit. Proses PCR dilakukan pada suhu tinggi dan memungkinkan terjadinya denaturasi enzim, sehingga diperlukan DNA polimerase yang termostabil dari bakteri termofilik. Penggunaan PCR yang meningkat mengakibatkan akibat masa pandemi mengakibatkan harga enzim tinggi. Sampai saat ini Indonesia masih menggunakan DNA polimerase dari impor sehingga menyebabkan tingginya biaya operasi. Di sisi lain, Indonesia memiliki potensi besar dalam pemanfaatan bakteri termofilik alami sehingga memungkinkan dilakukannya produksi DNA polimerase dari bakteri termofilik lokal seperti Geobacillus thermoleovorans strain SGAir0734 (GBK Pol) yang diisolasi dari sumber air panas Batu Kuwung, Banten. Sebelumnya, GBK Pol telah berhasil diproduksi pada skala laboratorium dalam flask 1000 ml. Pada penelitian ini, GBK Pol sekuens utuh diproduksi pada dalam flask yang diisi 50 ml, 100 ml, dan 250 ml serta pada skala bench menggunakan fermentor dengan volume kerja 3000 ml. GBK Pol berhasil dipurifikasi menggunakan metode immobilized metal affinity chromatography (IMAC), kemudian diuji menggunakan SDS-PAGE dan menghasilkan protein GBK Pol pada ±50 kDa. Kondisi optimum kultur pada fermentor mencakup: induksi IPTG pada jam ke-2 setelah kultur dan inkubasi setelah induksi selama 3 jam.

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DNA polymerase is a DNA synthesis enzyme that is utilized in DNA polymerase chain reaction (PCR) amplification method for disease detection. PCR process is carried out at high temperatures which allows denaturation of the enzyme, therefore, a thermostable DNA polymerase from thermophilic bacteria is required. The increased usage of PCR after the pandemic resulting in high enzyme prices. Until now, Indonesia still uses DNA polymerase from imports, causing high operating costs. On the other hand, Indonesia has great potential in utilizing naturally occurring thermophilic bacteria to produce DNA polymerase from local source such as the Geobacillus thermoleovorans strain SGAir0734 (GBK Pol) isolated from Batu Kuwung hot springs, Banten. Previously, GBK Pol had been successfully produced on a laboratory scale in 1000 ml flasks. In this study, a full sequence GBK Pol was produced with working volume of 50 ml, 100 ml and 250 ml in flasks and 3000 ml in a fermenter. GBK Pol was successfully purified using the immobilized metal affinity chromatography (IMAC) method, and tested using SDS-PAGE, resulting in GBK Pol protein at ±50 kDa. The optimum conditions for culture on the fermentor were as follows: induction of IPTG after 2 hours and 3 hours incubation after induction."

"Sistem CRISPR-Cas9 merupakan mekanisme perlindungan bakteri terhadap materi genetik asing yang diaplikasikan secara luas dalam rekayasa genetika. Kombinasi enzim Cas9 dengan gRNA pada CRISPR-Cas9 memungkinkan terjadinya pengeditan genom terhadap target yang spesifik. Meskipun demikian, Cas9 yang dikembangkan saat ini berasal dari bakteri mesofilik sehingga rentan terdegradasi dan tidak cocok untuk aplikasi pada temperatur tinggi. Di sisi lain, bakteri termofilik Geobacillus kaustophilus telah diisolasi dari mata air panas di Cisolong, Banten, dan diidentifikasi mengandung enzim Cas9. Untuk memperoleh enzim Cas9 yang tidak terdenaturasi pada temperatur tinggi (termostabil), dilakukan uji coba produksi Cas9 rekombinan dari Geobacillus kaustophilus. Gen Cas9 yang telah dikloning pada plasmid pET43.1a ditransformasikan ke dalam Escherichia coli BL21 dan dikultur dengan konsentrasi penambahan IPTG yang bervariasi—0,05 mM, 0,20 mM, dan 0,50 mM. Hasil kultur bakteri dipanaskan pada temperatur yang bervariasi (50 °C, 60 °C, dan 70 °C) untuk mendenaturasi enzim non-termofilik. Setelah itu, sampel dipurifikasi dengan mmobilized Metal Affinity Chromatography untuk memperoleh enzim Cas9. Hasil uji Lowry menunjukkan sampel heated supernatant dengan konsentrasi IPTG 0,05 mM dan temperatur pemanasan 50 °C memiliki konsentrasi protein tertinggi dan hasil purifikasinya memiliki konsentrasi Cas9 sebesar 42,6 μg/mL. Identifikasi protein dengan uji SDS-PAGE menunjukkan ukuran protein hasil purifikasi sebesar 52,61 kDa.

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The CRISPR-Cas9 system is a bacterial defense mechanism against foreign genetic material that is broadly applied in genetic engineering. The combination of Cas9 enzyme and gRNA in CRISPR-Cas9 allows genome editing of specific targets. However, the currently developed Cas9 originated from mesophilic bacteria, making it susceptible to degradation and unsuitable for applications requiring elevated temperatures. On the other hand, the thermophilic bacterium, Geobacillus kaustophilus, was isolated from a hot spring in Cisolong, Banten, and identified as containing the Cas9 enzyme. To obtain undenatured Cas9 enzymes at high temperatures (thermostable), a production test of recombinant Cas9 from Geobacillus kaustophilus was carried out. The Cas9 gene cloned on the pET43.1a plasmid was transformed into Escherichia coli BL21 and cultured under various IPTG addition concentrations—0.05 mM, 0.20 mM, and 0.50 mM. The bacterial cultures were heated at various temperatures (50 °C, 60 °C, and 70 °C) to denature unwanted non-thermophilic enzymes. Thereafter, the samples were purified using Immobilized Metal Affinity Chromatography to obtain Cas9 enzyme. Lowry protein assay results showed that the heated supernatant sample with 0.05 mM IPTG addition and 50 °C heating temperature has the highest protein concentration, and the purified sample yielded a Cas9 concentration of 42.6 μg/mL. Protein identification with SDS-PAGE revealed a purified protein size of 52.61 kDa"

"Structural Health Monitoring Photogrammetry & DIC, Volume 6: Proceedings of the 36th IMAC, A Conference and Exposition on Structural Dynamics, 2018, the sixth volume of nine from the Conference brings together contributions to this important area of research and engineering. The collection presents early findings and case studies on fundamental and applied aspects of Structural Health Monitoring & Damage Detection, including papers on; Structural Health Monitoring, Damage Detection, System Identification & Active Controls."

"Rotating Machinery, Vibro-Acoustics & Laser Vibrometry, Volume 7: Proceedings of the 36th IMAC, A Conference and Exposition on Structural Dynamics, 2018, the seveth volume of nine from the Conference brings together contributions to this important area of research and engineering. The collection presents early findings and case studies on fundamental and applied aspects of Rotating Machinery, Hybrid Testing, Vibro-Acoustics & Laser Vibrometry, including papers on; Rotating Machinery, Vibro-Acoustics, Experimental Techniques, and Scanning Laser Doppler Vibrometry Methods."

"DNA polimerase merupakan sebuah enzim yang memiliki aplikasi luas dalam dunia bioteknologi. Peran DNA polimerase dalam sintesis DNA membuat DNA polimerase sebuah reagen yang sering digunakan pada berbagai metode amplifikasi DNA. Proses sintesis DNA pada metode-metode tersebut menggunakan suhu tinggi, sehingga dibutuhkan DNA polimerase yang tidak terdenaturasi pada suhu tinggi atau dengan kata lain termostabil. Salah satu sumber DNA polimerase termostabil adalah bakteri termofilik Geobacillus thermoleovorans Strain SGAir0734 (GBK Pol) yang diisolasi dari mata air panas Batu Kuwung, Banten, Indonesia. Pada penelitian sebelumnya, telah diuji coba produksi dari GBK Pol dengan suhu purifikasi pada rentang 60 – 80℃ dimana diduga terjadi denaturasi dari enzim. Maka dari itu, pada penelitian ini dilakukan purifikasi GBK Pol sekuens utuh dan parsial dengan metode immobilized metal affinity chromatography (IMAC) dengan rentang suhu 40 – 60℃ dan uji aktivitas amplifikasi DNA dengan metode loop mediated isothermal amplification (LAMP) dengan rentang suhu 25 – 60℃. Hasil uji Lowry menunjukkan bahwa pemanasan 60℃ menghasilkan GBK pol dengan konsentrasi tertinggi, yaitu sekitar 134 µg/ml untuk plasmid sekuens utuh. GBK pol hasil purifikasi kemudian diuji aktivitasnya dengan LAMP, dimana reaksi pada suhu 40℃ selama 2 jam menghasilkan pita yang jelas pada elektroforesis dan menghasilkan konsentrasi DNA tertinggi. GBK pol dari hasil purifikasi dengan pemanasan 60℃ menghasilkan konsentrasi DNA tertinggi pada LAMP suhu 40℃ baik untuk sekuens parsial dan utuh.

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DNA polymerase is an enzyme that has a wide application in the world of biotechnology. DNA Polymerase’s role in synthesizing DNA makes DNA polymerase a favorable reagent in various DNA amplification methods. The synthesis process in these methods is done in high temperatures, thus a DNA polymerase that won’t experience denaturation in high temperatures, or in other words thermostable, is needed. One source of thermostable DNA polymerase is the thermophilic bacteria Geobacillus thermoleovorans Strain SGAir0734 (dubbed GBK Pol) that has been isolated from Batu Kuwung hot springs, Banten, Indonesia. In the previous study, a production trial of GBK pol has been carried out with purification temperatures that range between 60 – 80℃, where denaturation of the enzyme is suspected to have happened. As such, in this study, lower temperatures of 40 – 60℃ are employed in the purification of GBK Pol with immobilized metal affinity chromatography (IMAC) as well as in the activity study with loop mediated isothermal amplification (LAMP) with reaction temperatures of 25 – 60℃. The Lowry assay results show that the highest GBK pol concentration is achieved with 60℃ heat treatment, with a concentration of around 134 µg/ml for full sequence plasmid. Purified GBK pol are then tested with LAMP, where isothermal amplification at 40℃ for 2 hours resulted in the clearest bands during gel electrophoresis as well as highest concentrations of DNA. The highest concentration of DNA is achieved from GBK pol with 60℃ heat treatment, suggesting that 60℃ is the optimal temperature for purification."

"Nonlinear Dynamics, Volume 1: Proceedings of the 36th IMAC, A Conference and Exposition on Structural Dynamics, 2018, the first volume of nine from the Conference brings together contributions to this important area of research and engineering. The collection presents early findings and case studies on fundamental and applied aspects of Nonlinear Dynamics, including papers on; Nonlinear System Identification, Nonlinear Modeling & Simulation, Nonlinear Reduced-order Modeling, Nonlinearity in PracticeNonlinearity in Aerospace Systems, Nonlinearity in Multi-Physics Systems, Nonlinear Modes and Modal Interactions, and Experimental Nonlinear Dynamics."