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Abstrak :
Enzim adalah suatu protein yang bekerja sebagai katalisator organik, mengatur reaksi-reaksi kimia dalam setiap organisme (1). Enzim-enzim yang sepanjang waktu terdapat di dalam plasma dan melakukan fungsi fisiologiknya dalam plasma dikenal sebagai enzim khas plasma seperti lipase lipoprotein, pseudokolinesterase dan proenzim-proenzim untuk pembekuan darah. Enzim-enzim tersebut di atas umumnya disintesis di dalam hati tetapi terdapat dalam darah dengan konsentrasi yang sama atau lebih tinggi dibandingkan konsentrasinya di dalam jaringan (2).Enzim-enzim plasma yang tidak melakukan fungsi fisiologiknya di dalam plasma dikenal sebagai enzim tidak khas plasma. Enzim-enzim ini terdapat di dalam sel organ atau jaringan tertentu, dan dalam keadaan normal hanya sejumlah kecil yang ada dalam plasma. Bila terjadi kerusakan organ atau jaringan, aktivitas enzim-enzim ini di dalam plasma akan meningkat melebihi keadaan normal. Kenaikan aktivitas enzim-enzim ini di dalam plasma selain tergantung pada konsentrasinya di dalam jaringan, juga pada luas organ yang rusak dan lokasi enzim di dalam sel (2,3). Contoh enzim tidak khas plasma yaitu enzim fosfatase asam, enzim

Abstrak :
Open mining activities result in decreased microbial biomass and negatively impacts soil fertility. Soil microbes play a role in the decomposition of soil organic matter and in nutrient cycles through the process of mineralization by the enzymes they produce. The purpose of this study was to analyse soil fertility levels in PT Bukit Asam’s various reclaimed land sites at Muara Enim Regency, South Sumatra, Indonesia, as determined by these areas’ microbial populations and soil enzyme activity. The research was conducted by using the explorative method in PT Bukit Asam’s various reclaimed land. Soil sample from 7 different reclamation age area were analysed. Our results showed that soil conditions and soil enzyme activity vary by reclamation age. At KTU, a 12-year-old reclaimed land site, urease enzyme activity had a value of 68.83 mg NH4+.g-1dm.h-1 with a microbial population of 82.64 x 104 CFU.g-1soil. The highest phosphatase enzyme activity value of 95.66 mg pNP.g-1 dm.h-1 was found on the 9-year-old SP702 reclaimed land site, with a soil pH of 5.23. Cellulase enzyme activity on the 21-year-old Udongan reclaimed site had a value of 21.51 mg GE.g-1dm.h-1 with a cellulolytic microbial population of 1.9 x 104 CFU.g-1soil, higher than on other reclamation sites. Invertase enzyme activity on the 15-year-old Tupak reclaimed land site had a value of 24.37 mg GE.g-1dm.h-1. Soil enzyme activity can be an indicator of soil quality and soil microbial activity as it relates to all forms of biochemical transformations occurring in the soil and is highly sensitive to environmental changes.

Abstrak :
Bonggol nanas merupakan limbah tanaman nanas yang banyak mengandung zat aktif  potensial, yaitu bromelain. Bromelain merupakan enzim proteolitik yang memiliki efek terapetik sebagai agen antiinflamasi dan debridment enzimatik. Pemberian bromelain dalam basis topikal telah dilaporkan aman dan efektif untuk anti-inflamasi. Pada beberapa tahun terakhir ini, sistem penghantaran obat topikal yang cukup inovatif adalah nanoemulsi. Keunggulan daripada sediaan nanoemulsi adalah ukuran partikel yang rendah akan memudahkan kontak antara kulit dan sediaan, sehingga meningkatkan kemampuan zat aktif terpenetrasi. Maka dari itu, penelitian ini bertujuan untuk memformulasikan enzim bromelain hasil isolasi parsial dari bonggol nanas dalam sediaan serta uji penetrasi menggunakan alat seldifusi Franz. Isolasi dan purifikasi bromelain dilakukan dengan fraksionasi bertingkat menggunakan garam ammonium sulfat dan dialisis. Fraksi bromelain terbaik pada tingkat kejenuhan garam ammonium sulfat 20-50% yaitu sebesar 70,68 U/mg kemudian fraksi tersebut didialisis mengalami peningkatan aktivitas enzim sebesar 135,92 U/mg. Basis nanoemulsi optimum pada konsentrasi surfaktan Tween 80 sebesar 30%.  Nanoemulsi memiliki karakteristik minyak dalam air (m/a), Ukuran globul sebesar 21,37nm dengan indeks polidispersitas (pdI) sebesar 0,323. Kestabilan basis nanoemulsi diamati sampai minggu ke-8 dengan hasil tampak jernih, dan tidak terjadi pemisahan fase.

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Pineapple core is a waste of pineapple plants which contain many  potential active substances, one of them is called bromelain. Bromelain is a proteolytic enzyme that has a therapeutic effect as an anti-inflammatory and enzymatic debridment agent. The administration of bromelain in a topical base topically has been reported to be safe and effective for anti-inflammation. In recent years, the most innovative topical drug delivery systems have been nanoemulsion. The advantage of nanoemulsion base is low particle size will easily contact between the skin and the base, so that will increase the ability of penetration of active substance. Therefore, this study aims to formulate the bromelain enzyme resulting from partial isolation from pineapple core in the nanoemulsion preparation and penetration test using Franz diffuse cell. Isolation and purification of bromelain was carried out by stratified fractionation using ammonium sulfate salt and dialysis. The best bromelain fraction at the saturation level of ammonium sulfate salt was 20-50% which was 70.68 U/mg then the dialyzed fraction experienced an increase in enzyme activity of 135.92 U/mg. The optimum nanoemulsion base at the concentration of Tween 80 surfactant was 30%. Nanoemulsion has the characteristics of oil in water (m/a), globule size of 21.37nm with polydispersity index (pdI) of 0.323. The stability of the nanoemulsion base was observed until the 8th week with the results appearing clear, and no phase separation occurred.

Abstrak :
Breast cancer stem cells (BCSCs) dengan petanda Aldehida dehidrogenase 1-positif (ALDH1+) merupakan populasi minor dari sel-sel tumor dengan kemampuan tumorigenik yang tinggi dan bertahan terhadap stres oksidatif. Manganese superoksida dismutase (MnSOD) merupakan pertahanan utama terhadap superoksida yang diekspresikan spesifik di mitokondria, yang merupakan salah satu sumber utama stres oksidatif di dalam sel.  Sejauh ini, belum diketahui peranan MnSOD terhadap ketahanan hidup dan kepuncaan BCSC. Transfeksi in vitro pada BCSC (ALDH1+) dilakukan dengan menggunakan siRNA MnSOD spesifik dalam kondisi kultur standar. Total RNA dan protein diekstraksi dengan menggunakan TriPure® Isolation Reagent dan RIPA® lysis buffer. Viabilitas sel diukur dengan menggunakan trypan exclusion assay. Ekspresi relatif mRNA MnSOD dan OCT4 dianalisis dengan menggunakan one-step qRT-PCR. Aktivitas MnSOD diukur dengan menggunakan uji inhibisi xantin oksidase (RanSOD® kit). Kadar superoksida sel diukur dengan menggunakan uji dihidroetidium dan tumorigenik diukur dengan menggunakan mammosphere-forming unit. Setelah diinkubasi selama 48 jam dengan menggunakan siRNA dengan menggunakan dosis 80 pmol. Ekspresi relatif mRNA MnSOD mengalami penekanan sejumlah 0,17-kali (p<0,01), penurunan aktivitas spesifik MnSOD sebesar 70,4 %, peningkatan kadar superoksida sel menjadi 1,13-kali, penurunan ekspresi OCT4 menjadi 1,08-kali (p<0,05) dan penurunan mamosphere forming unit efficiency menjadi 36,5 % (p<0,05) dibandingkan dengan kontrol negatif. Viabilitas BCSC (ALDH1+) menurun sebanyak 75 %(p<-0,05) dibandingkan kontrol negatif. Hasil penelitian ini menunjukkan bahwa penekanan ekspresi MnSOD dapat menjadi target yang menjanjikan untuk menurunkan kepuncaan dan tumorigenitas BCSC (ALDH1+). ......Aldehyde dehydrogenase 1-positive (ALDH1+) breast cancer stem cells (BCSCs) are a small population of tumor cells with high capacity of tumorigenicity and oxidative stress. Manganese superoxide dismutase (MnSOD) is specifically expressed in mitochondria as the primary defense against superoxides, which are one of the causes of oxidative stress in cells. The aim of this study was to determine the impact of suppressing MnSOD expression using small interfering RNA (siRNA) on the stemness, tumorigenicity, and viability of BCSCs. In vitro transfection of ALDH1+ BCSCs was performed using 33 and 66 µM specific MnSOD siRNA under standard culture conditions. Total RNA and protein were extracted from the transfected cells using TriPure® Isolation Reagent and RIPA® lysis buffer. Cell viability was measured using a trypan blue exclusion assay. The relative expression of MnSOD and OCT4 mRNAs was analyzed by one step qRT-PCR. MnSOD activity was determined by xanthine oxidase inhibition assay (RanSOD® kit). Cellular superoxides were measured using a dihydroethidium assay and tumorigenicity was observed with mammosphere-forming unit.  After siRNA incubation for 48 hours, MnSOD was suppressed by 0.176-fold (p<0.01), MnSOD enzyme specific activity was reduced 70.4%, cellular superoxide levels increased by 1.13-fold, OCT4 expression was suppressed by 1.98-fold (p<0.05), and mammosphere-forming unit decreased by 36.5% (p<0.05) compared with the corresponding negative controls. The viability of the ALDH1+ BCSCs was reduced 75% (p< 0.05). Our results suggest that suppression of MnSOD expression may be a promising target to reduce stemness and tumorigenicity of ALDH1+ BCSCs.

Abstrak :
The purpose of this study was to get optimum medium composition and agitation to trypsin-like protease production by Lactobacillus plantarum FNCC 0270. The medium composition and agitation for enzyme production was optimized using Central Composite Design and Response Surface Method with Design Expert software version 7.1.5. Fermentation was carried out in erlenmeyer flask at initial pH 8, 37 °C, with shaker incubator at 87.5 rpm. The results of the best of enzyme activity 1.0 mU/mL, protein levels of 0.557 mg/mL and desirability value of 0.740. Numerical optimization was performed to approach the ideal state of the fermentation or desirability value of 1. The medium composition of fermentation used was: 3.64% baker's yeast, 1.21% glucose, and 0.13% skim milk. The enzyme activity reached was 1.51 mU/mL and protein levels of 0.205 mg/mL. After numerical optimization, the fermentation process was verified using 125 mL Erlenmeyer in shaking incubator at 77 rpm, initial pH 8, 37 °C, 15 h of fermentation. The verification results showed that the enzyme activity and protein levels was 1.273 ± 0.227 mU/mL and 0.248 ± 0.012 mg/mL, respectively.

Optimization of Trypsin-like Protease Production by Lactobacillus plantarum FNCC 0270 using Response Surface Methodology. Tujuan penelitian ini adalah memperoleh komposisi medium dan agitasi optimum untuk produksi protease serupa tripsin oleh Lactobacillus plantarum FNCC 0270. Protease serupa tripsin (PST) dihasilkan oleh L. plantarum FNCC 0270 melalui proses fermentasi dengan optimasi komposisi medium dan agitasi menggunakan Central Composite Design dan Response Surface Methode, dengan software Design Expert versi 7.1.5. Fermentasi dilakukan dalam erlenmeyer pH awal 8 dan suhu 37 °C menggunakan shaker inkubator pada 87,5 rpm. Hasil eksperimen terbaik menunjukkan aktivitas enzim 1 mU/mL, kadar protein 0.557 mg/mL dan diperoleh nilai desirability 0,740. Untuk mendekati keadaan ideal atau nilai desirability 1 dilakukan optimasi melalui simulasi numerik, yaitu fermentasi dengan komposisi baker yeast 3,64%, kadar glukosa 1,21%, konsentrasi skim milk 0,13%, dan agitasi 77 rpm, sehingga diperoleh aktivitas enzim 1,51 mU/mL dan kadar protein 0,205 mg/mL. Setelah optimasi numerik kemudian dilakukan verifikasi fermentasi di laboratorium, dalam erlenmeyer menggunakan shaker inkubator, agitasi 77 rpm, pH awal 8, suhu 37 °C, selama 15 jam. Hasil verifikasi menunjukkan aktivitas enzim dan kadar protein masing-masing 1,273 ± 0,227 mU/mL dan 0,248 ± 0,012 mg/mL.