Hasil Pencarian  ::  Simpan CSV :: Kembali

Abstrak :
Ruang Lingkup dan Cara Penelitian: Peneliti di Indonesia sering mengalami kesulitan memperoleh bahan baku seperti petanda protein untuk menentukan 'berat molekul'. Salah satu petanda protein adalah inhibitor tripsin kacang kedelai (ITK). Penelitian ini bertujuan mengisolasi dan memurnikan ITK dari biji kacang kedelai (Glycine max), Isolasi dilakukan dengan cara ekstraksi asam, diikuti 'salting out, dialisis dan pengendapan aseton. Pemurnian dilakukan dengan kromatografi kolom pertukaran ion. Aktivitas ITK fraksi 'kasar' dan 'murni' diperiksa dengan mengamati pengaruh hambatan terhadap aktivitas enzimatik tripsin, dengan substrat azokasein (proteolitik) dan BAPA/BAPNA (amidase). Azokasein yang dipakai sebagai substrat reaksi tripsin disintesis sendiri. Penilaian kemurnian ITK ?kasar? dan 'murni', juga tripsin dilakukan dengan elektroforesis gel 'slab' SDS-poliakrilamid. Sebagai pembanding pada penelitian ini digunakan inhibitor tripsin produk Sigma. Hasil dan Kesimpulan : Dari 100 g biji kacang kedelai kering panen, diperoleh 1,16 g isolat (ITK 'kasar') setara dengan 418,46 mg protein. Pemurnian lewat kolom pertukaran ion menghasilkan 2 fraksi dengan 'recovery' protein total minimal 63,7 %. Azokasein yang disintesis hasilnya berbeda bila jenis alkohol yang digunakan juga berbeda. Pada penelitian ini baik ITK 'kasar` maupun 'murni' memperlihatkan hambatan terhadap reaksi enzimatik tripsin. Hambatan 50 % terjadi pads rasio inhibitor/enzim yang bervariasi; ITK `murni' 1 memperlihatkan angka yang paling tinggi. Elektroforetogram menunjukkan bercak ITK 'murni' II identik dengan SBTI (Sigma).

---

Scope and Method of Study: The chemicals required for laboratory investigations are quite often difficult to obtain, e. g. the standard protein markers for molecular weight determination. Among the markers used for that purpose is the soybean trypsin inhibitor. This work was carried out to isolate and purify trypsin inhibitor from soybean (Glycine max) seeds (SBTI). The procedure included an acid extraction, followed by salting-out, dialysis and an acetone precipitation. Purification was carried out by ion-exchange column chromatography. The protein content of the isolate and of the purified substance was determined by spectrophotometer. The activities of the crude and of the purified soybean trypsin inhibitor were tested against trypsin activity. The trypsin used was obtained commercially. Trypsin's proteolytic activity was performed on azo-casein while its amidase activity was tested on BAPA/BAPNA. The azo-casein was synthesized in the laboratory. The purity of the crude and of the purified soybean trypsin inhibitors, and of the trypsin itself was examined on a slab SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Findings and Conclusions: From 100 g of dried soy-bean seeds, 1.16 of product was isolated (= 418.46 mg protein). Further purification on an ion-exchange column yielded two fractions, with a minimum of 63.7% protein recovered. The azo-casein synthesis revealed two different products depending on the grade of alcohol used in the process. The crude and the purified soybean trypsin inhibitors showed inhibitory effects towards trypsin. The fifty percent inhibition occurred at varied inhibitor/enzyme ratios, the highest was shown by the purified SBTI I. The electrophoretogram showed that the purified SBTI II was identical to the SBTI (Sigma).

Abstrak :
Penelitian penapisan beberapa Lactobacillus plantarum dan optimasi produksi protease serupa tripsin (PST) dilanjutkan dengan pemekatan dan karakterisasi parsial telah dilakukan. Tripsin memiliki peran penting dalam pencernaan protein di usus kecil namun produksi tripsin komersial saat ini masih terbatas oleh masalah sertifikasi halal dan risiko penularan penyakit yang bersumber dari babi atau sapi. Penelitian bertujuan menyeleksi koleksi isolat L. plantarum yang menghasilkan aktivitas PST tertinggi dan menentukan kondisi optimum dalam produksi PST dari L. plantarum terpilih menggunakan Response Surface Methodology (RSM) diikuti dengan pemurnian dan karakterisasi parsialnya. Tujuh isolat L. plantarum yang diperoleh dari makanan tradisional Indonesia diseleksi secara kualitatif dan kuantitatif. Semua isolat L. plantarum menunjukkan aktivitas proteolitik, terlihat adanya zona bening di sekitar koloni. Zona bening menunjukkan adanya potensi L. plantarum sebagai sumber PST secara kualitatif. Hasil pengujian kuantitatif menunjukkan bahwa isolat dengan kode B6 (L. plantarum WBM-4) menghasilkan PST dengan aktivitas tripsin tertinggi sebesar 0,16 mU/mL. Lactobacillus plantarum WBM-4 diisolasi dari buah Menteng Banjarmasin paling berpotensi untuk menghasilkan PST. Selanjutnya, L. plantarum WBM-4 dioptimalisasi produksi menggunakan Respon Surface Methodology (RSM) dan karakterisasi PST. Kondisi optimal ditentukan pada komposisi medium dengan 1,96% glukosa, 0,39% yeast extract, 1,97% skim milk, dan pH 6,62, menghasilkan aktivitas PST sebesar 0,303 mU/mL. Pemekatan enzim kasar di bawah kondisi optimum menggunakan viva spin 5000 MWCO meningkatkan kemurnian hingga 11,08 kali lipat, dengan aktivitas sebesar 2,47 mU/mL. Karakterisasi parsial menunjukkan berat molekul PST sekitar ~19 kDa dan ~29 kDa, stabilitas dalam rentang suhu 30 - 40°C, dan aktivitas optimal pada pH 7,0 - 8,0. Penambahan ion logam EDTA, Ca2+, dan Zn2+ memengaruhi aktivitas PST. Penyimpanan PST selama 30 hari pada suhu 4°C aktivitas tersisa PST masih 65% sedang pada suhu 24-28°C aktivitas hanya tersisa 15%. Hasil penelitian ini memberikan gambaran tentang potensi PST yang berasal dari L. plantarum untuk aplikasi suplemen pencernaan dan memberikan alternatif sumber tripsin yang halal dan aman. ......Research on screening of several Lactobacillus plantarum and optimization of trypsin-like protease production (TLP) followed by concentration and partial characterization has been carried out. Trypsin has an important role in protein digestion in the small intestine, but commercial trypsin production is currently limited by halal certification issues and the risk of transmission of diseases sourced from pigs or cattle. The study aimed to select a collection of Lactobacillus plantarum isolates that produced the highest TLP activity and determine the optimum conditions in TLP production from selected L. plantarum using Response Surface Methodology (RSM) followed by purification and partial characterization. Seven isolates of L. plantarum obtained from traditional Indonesian food were selected qualitatively and quantitatively. All L. plantarum isolates exhibited proteolytic activity, with clear zones around the colony. The clear zone shows the potential of L. plantarum as a qualitative source of TLP. The results of quantitative testing showed that isolates with code B6 (L. plantarum WBM-4) produced TLP with the highest trypsin activity value 0.16 mU/mL. L. plantarum WBM-4 isolated from Banjarmasin Menteng fruit has the most potential to produce TLP. Furthermore, L. plantarum WBM-4 optimized production using Response Surface Methodology (RSM) and TLP characterization. Optimal conditions were determined in the composition of the medium with 1.96% glucose, 0.39% yeast extract, 1.97% skim milk, and pH 6.62, resulting in TLP activity of 0.303 mU/mL. Crude enzyme concentration under optimum conditions using viva spin 5000 MWCO increases purity up to 11.08-fold, with an activity of 2.47 mU/mL. Partial characterization shows TLP molecular weights of approximately ~29 kDa and ~19 kDa, stability in the temperature range of 30 - 40 °C, and optimal activity at pH 7.0 - 8.0. The addition of EDTA, Ca2+, and Zn2+ metal ions affect TLP activity. TLP storage for 30 days at 4°C the remaining activity of PST is still 65% while at 24-28°C the activity is only 15%. The results of this study provide an overview of the potential of PST derived from L. plantarum for digestive supplement applications and provide an alternative source of trypsin that is halal and safe.

Abstrak :
ABSTRAK
Diare persisten merupakan masalah kesehatan serius dan sering menyebabkan malnutrisi. Kerusakan mukosa pada diare diduga menyebabkan penurunan hormon sekretin dan kolesistokinin sehingga mengurangi stimulasi ke pankreas dan memperberat diare persisten dan malnutrisi.Penelitian ini bertujuan untuk mengetahui fungsi eksokrin pankreas pada anak diare persisten, anak malnutrisi, mendapatkan nilai referensi pemeriksaan fecal elastase-1 FE-1 anak Indonesia, dan mengetahui kehandalan analisis feses dan steatokrit dalam mendeteksi insufisiensi eksokrin pankreas.Penelitian potong lintang pada tahap pertama dilakukan untuk mendapatkan sebaran nilai FE-1 pada anak normal, membandingkan nilai FE-1 subjek diare persisten dan malnutrisi dengan anak normal, dan mengetahui sensitivitas, spesifisitas, dan kemampuan diskriminasi analisis feses dan steatokrit dalam mendeteksi insufisiensi eksokrin pankreas. Tahap kedua uji klinis dua kelompok paralel tersamar ganda dilakukan untuk menguji efek suplementasi enzim pankreas 8371 USP unit tiga kali sehari selama sebulan pada anak diare persisten. Penelitian dilakukan di 5 Rumah Sakit di Jakarta Januari 2015 minus;Juli 2016 pada anak berusia 6 ndash;60 bulan.Sebanyak 182 anak usia 6 ndash;60 bulan direkrut sebagai subjek yang terdiri dari 31 anak dengan diare persisten, 31 anak dengan malnutrisi, dan 120 anak normal. Nilai cut-off FE-1 yang didapatkan pada penelitian ini adalah 307 mcg/g feses. Terdapat perbedaan bermakna nilai FE-1 antara subjek diare persisten dan anak normal. Tidak ditemukan perbedaan bermakna nilai FE-1 antara subjek malnutrisi dan anak normal. Terdapat perbedaan bermakna lama diare sekitar 7 hari antara kedua kelompok. Kadar FE-1 dan prealbumin antara baseline dan endpoint pada kelompok plasebo dan perlakuan tidak berbeda bermakna. Uji kehandalan masing-masing komponen analisis feses dan steatokrit menunjukkan hasil sensitivitas dalam rentang 5 ndash;32 , spesifisitas 73 ndash;98 , nilai prediksi positif 1 ndash;43 , dan nilai prediksi negatif 87 ndash;89 . Nilai AUC analisis feses dan steatokrit masing-masing adalah 0,664 IK 95 0,539 ndash;0,788 dan 0,501 IK 95 0,372 ndash;0,629 sedangkan AUC gabungan sebesar 0,671.Kesimpulannya, pada penelitian ini didapatkan adanya insufisiensi eksokrin pankreas pada anak dengan diare persisten. Suplementasi enzim pankreas terbukti dapat memperpendek lama diare secara bermakna. Analisis feses dan/atau steatokrit memiliki sensitivitas yang rendah, spesifisitas yang tinggi, dan kemampuan diskriminasi kurang.Kata kunci: anak, diare persisten, fungsi eksokrin pankreas, malnutrisi, suplementasi enzim pankreas

---

ABSTRACT
Persistent diarrhea is a serious health problem and is closely related to malnutrition. Prolonged mucosal injury in diarrhea is thought to cause reduced secretin and cholecystokinin CCK secretion, which decreases stimulation to the pancreas and further aggravate persistent diarrhea and malnutrition.This research aims to study pancreatic exocrine function in children with persistent diarrhea and children with malnutrition, to obtain reference values of fecal elastase 1 FE 1 in Indonesian children, and to assess the ability of stool analysis and steatocrit in detecting exocrine pancreatic insufficiency.Cross sectional study was done to obtain FE 1 distribution in healthy children, to study FE 1 levels in children with persistent diarrhea and children with malnutrition, and to study the sensitivity, specificity, and discriminative capacity of stool analysis and steatocrit in detecting exocrine pancreatic insufficiency. A randomized, two double blind parallel group, placebo controlled clinical trial was conducted to evaluate the effects of 8371 USP units of pancreatic enzyme replacement therapy PERT 3 times daily for 1 month in children with persistent diarrhea. This study involved children age 6 ndash 60 months in 5 hospitals in Jakarta from January 2015 to July 2016.As much as 182 children 6 ndash 60 months of age consisting of 31 children with persistent diarrhea, 31 children with malnutrition, and 120 healthy children were recruited as subjects. Cut off point of FE 1 in this study was 307 mcg g faeces. Significant difference of FE 1 was found between children with persistent diarrhea and healthy children. The FE 1 difference between subjects with malnutrition and healthy children was not significant. Duration of diarrhea was 7 days significantly shorter in the PERT group. Changes of FE 1 and prealbumin values between baseline and endpoint in placebo and treatment group were found to be statistically insignificant. The diagnostic value of each stool analysis component and steatocrit test showed that the sensitivity was within range of 5 ndash 32 , specificity 73 ndash 98 , positive predictive value 1 ndash 43 and negative predictive value 87 ndash 89 . The AUC values of stool analysis and steatocrit were 0.664 95 CI 0.539 ndash 0.788 and 0.501 95 CI 0.372 ndash 0.629 , respectively, and the combined AUC 0,671.In conclusion, exocrine pancreatic insufficiency was observed in children with persistent diarrhea, and PERT has been proven to significantly shorten the duration of diarrhea by 1 week. Stool analysis and or steatocrit has low sensitivity, high specificity, and low discrimination capacity.Keywords children, exocrine pancreatic function, malnutrition, pancreatic enzyme supplementation, persistent diarrhea