Optimum condition for Nigella sativa seeds oil and its bioactive compound, thymoquinone (TQ) using supercritical fluid carbon dioxide extraction (SCFE-CO 2 ), were investigated. The optimization process was performed with Taguchi method and full factorial design (FFD) under the following condition: pressure (150, 200 and 250 bar), temperatures (40, 50 and 60 o C) and carbon dioxide (CO 2 ) flowrate (10, 15 and 20 g/min), in which solvent to feed (SF) ratio was set constant at 24. The highest yield of Nigella sativa seeds oil from SCFE-CO 2 process with FFD method was 12% at 250 bar, 60 o C and 20 g/min. Whereas Taguchi method was performed at 250 bar, 50 o C, 10 g/min with oil yield 11.9%. The highest thymoquinone content in Nigella sativa seeds oil from both experimental design was achieved through SCFE-CO 2 extraction condition at 150 bar, 60 o C and 20 g/min with thymoquinone content 20.8 mg/ml. In addition, conventional methods such as high pressure soxhlet with liquid CO 2 , n-hexane soxhlet and percolation with ethanol were performed with oil yield 5.8%, 19.1% and 12.4%; and thymoquinone content 8.8 mg/g oil, 6.3 mg/g oil and 5.0 mg/g oil, respectively. The analysis of variance (ANOVA) with 95% confidence interval, indicates effect of pressure on Nigella sativa seeds oil yield and thymoquinone content from SCFE-CO 2 process. The oil was then evaluated for its activity. The antibacterial activity of the oil from Taguchi Method, shows that all samples were unable to inhibit Escherichia coli O157 and Salmonella typhimurium. For other pathogenic bacterias, all samples show similar inhibition at concentration 10% oil for MRSA, 3% oil for Stapylococcus aureus and 3% oil for Bacillus subtilis.

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The anti-hypercholesterolemic effect of 11 Lactobacillus isolates was investigated in vitro by measuring remaining cholesterol in growth media, growth ability in media supplemented with cholesterol, and BSH activity. Among the selected isolates, DLBSH104, DLBSH122, and DLBSK207 have demonstrated outstanding potential as cholesterol-lowering cultures. The three isolates showed high cholesterol removal by growing cells, whereas resting and dead cells showed less cholesterol removal. Furthermore, visualization of those isolates in growing and non-growing states by SEM showed the ability of DLBSH104 to attach cholesterol to their cell surface. In contrast, alteration of DLBSH122 and DLBSK207 cells did not involve surface attachment of cholesterol. Thus, the isolates’ ability to remove cholesterol is mainly attributed to the cells’ metabolically active state that assimilates and incorporates cholesterol into the cell membrane as reflected by a significantly higher cholesterol removal in a growing state than a non-growing state. Only in DLBSH104 did cholesterol removal also involve attachment on the cell surface. Moreover, DLBSH104 has beneficially affected the host cell by a significant reduction of NPC1L1 mRNA levels that are responsible for intestinal cholesterol absorption. In hepatic cells, cell-free supernatant (CFS) from DLBSH104 and DLBSK207 were able to reduce LDLR and HMGCR mRNA at the transcription level. To sum up, L. helveticus DLBSH104 and L. plantarum DLBSK207 are confirmed as isolates with an anti-hypercholesterolemic effect.

Viability of Lactobacillus acidophilus DLBSD102 after Microencapsulation      This study was aim to select the viability the Lactobacillus acidophilus DLBSD102 during the spray drying method, to produce a fermented milk powder containing probiotic. Since spray drying process use the high temperature, suitable encapsulation material will increase the vaibility of probiotic and the quality of the final product. Three different encapsulation materials which were maltodextrin, whey protein isolate, and inulin with several formulations were used. The spray drying temperature used in this study was 130°C (inlet) and 60°C (outlet). The quality of the fermented milk powder containing L.acidophilus DLBSD102 bacteria strain was evaluated by measure the bacterial viability, bacterial cell resistance from hot temperatures, bile salts (0.5%) low pH (pH 2.0), and the presence of possible pathogenic bacteria. The results showed that the additional encapsulation material of inulin yielded a good quality fermented milk powder, compared with a mixture of encapsulation materials of maltodextrin: whey protein isolate (3:1), based on viability of probiotics after spray drying was increased, bacterial cell resistance to hot temperature, bile salt (0.5%) low pH (pH 2.0), and resistance to the presence of pathogenic bacteria. The addition of encapsulation material in the form of inulin yielded viability of BAL bacteria with log decrease of 0.20 ± 0,01 log CFU/g whereas without inulin addition decreased by 0.51± 0.36 log CFU/g when dried. Therefore, the mixture of encapsulation materials :maltodextrin:whey protein isolate:inulin (3:1:1) is used in the microencapsulation process of BAL by yielding 8.93% heat resistance, bile salt resistance of 78.55%, resistance to pH 2 of 77.25%, total titrated acids by 2.38%, moisture content during storage of 4.33% (4°C) and 3.96% (25°), pH value during fermentation process was 3.59±0,35 and no pathogenic bacteria was detected during production, packaging and storage for 4 weeks.Keywords: L. acidophilus DLBSD102, microenkapsulation, enkapsulation material, spray dryingABSTRAK      Penelitian ini tentang viabilitas Lactobacillus acidophilus DLBSD102 menggunakan bahan enkapsulan yang sesuai dengan metode pengeringan semprot. Tujuannya menghasilkan sediaan produk probiotik berupa serbuk susu fermentasi. Efektivitas mikroenkapsulasi dapat ditingkatkan dengan pemilihan jenis bahan enkapsulan yang tepat saat akan dikeringkan. Suhu pengeringan semprot yang digunakan dalam penelitian ini adalah 130°C (inlet) dan 60°C (outlet). Bahan enkapsulan yang digunakan adalah campuran dari maltodekstrin:whey protein isolate:inulin (3:1:1). Kualitas serbuk susu fermentasi dari strain bakteri L.acidophilus DLBSD102 yang diperoleh dievalusi termasuk viabilitas bakteri, ketahanan sel bakteri terhadap suhu panas, garam empedu (0,5%) pH rendah (pH 2,0) dengan metode cawan tuang, dan evaluasi kemungkinan adanya bakteri patogen. Hasil penelitian menunjukkan bahwa bahan enkapsulan tambahan berupa inulin menghasilkan serbuk susu fermentasi dengan kualitas yang baik, dibandingkan dengan campuran bahan enkapsulan berupa maltodekstrin:whey protein isolate (3:1), yang didasarkan pada viabilitas probiotik setelah pengeringan semprot dan meningkatkan, ketahanan sel bakteri terhadap suhu panas, garam empedu (0,5%) pH rendah (pH 2,0), dan ketahanan terhadap adanya bakteri patogen. Penambahan bahan enkapsulan berupa inulin menghasilkan viabilitas bakteri BAL dengan log penurunan sebesar 0,20±0,01 log CFU/g sedangkan tanpa penambahan inulin mengalami penurunan sebesar 0,51±0,36 log CFU/g saat dikeringkan. Oleh sebab itu, campuran bahan enkapsulan maltodekstrin:whey protein isolate:inulin (3:1:1) digunakan dalam proses mikroenkapsulasi BAL dengan menghasilkan ketahanan terhadap panas sebesar 8,93%, ketahanan terhadap garam empedu sebesar 78,55%, ketahanan terhadap pH 2 sebesar 77,25%, total asam tertirasi sebesar 2,38%, kadar air selama penyimpanan sebesar 4,33% (4°C) dan 3,96% (25°), nilai pH selama proses fermentasi sebesar 3,59±0,35 dan serbuk susu fermentasi tidak mengandung bakteri patogen selama proses produksi, pengemasan hingga penyimpanan selama 4 minggu.Kata kunci: Probiotik L. acidophilus DLBSD102, mikroenkapsulasi, bahan enkapsulan