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"Sukrosafosforilase (SPase) merupakan suatu enzim yang dapat mengkatalisis reaksi pemindahan gugus glukosil dari molekul donor ke suatu molekul aseptor (glukosilasi). Glukosilasi telah banyak dimanfaatkan, terutama untuk meningkatkan stabilitas dan karakteristik suatu senyawa bioaktif. Pada penilitian ini dilakukan isolasi SPase dari bakteri Escherichia coli BL-21 STARTM rekombinan yang membawa gen penyandi sukrosafosforilase asal Leuconostoc mesenteroides MBFWRS-3(1). Uji konfirmasi berat molekul enzim telah berhasil dilakukan dengan SDS PAGE dan ditunjukan bahwa berat molekul SPase rekombinan berkisar antara 45?66 kDa, hal tersebut sesuai dengan studi sebelumnya. Aktivitas enzim diketahui dengan metode spektrofotometri dan didapatkan bahwa aktivitas relatif SPase rekombinan sebesar 98,5%. Esei aktivitas transglikosilasi SPase rekombinan terhadap substrat asam kojat telah berhasil dilakukan. Berdasarkan pengamatan KLT Densitometer, didapatkan bahwa produk transglikosilasi hasil esei aktivitas transglikosilasi SPase rekombinan dan SPase standar terhadap asam kojat memiliki kemiripan.

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Sucrose Phosphorylase (SPase) is an enzyme that catalyzes glucosyl transfer reaction from donor molecules to acceptor molecules (glucosylation). Glucosylation has been used for many things, especially to increase chemical stability and improving characteristic of several bioactive compounds. In this study SPase has isolated from Escherichia coli BL-21 STARTM recombinants that carried gene of SPase expression from Leuconostoc mesenteroides MBFWRS-3(1). Confirmation of molecular weight has done by SDS PAGE and showed that the molecular weight of SPase was in range 66?45 kDa, as reported in other existed SPase studies. The activity enzyme obtained by using the spectrophotometric method, and performed relative activity 98.5 %. Transglucosylation activity assay of SPase recombinant has done to kojic acid. Based on TLC Densitometry analyzes, transglucosylation product of SPase recombinant was similarly to transglucosylation product of SPase standart."

"Sukrosafosforilase (SPase) adalah suatu enzim yang mengkatalisis sejumlah reaksi pemindahan gugus glukosil ke sejumlah molekul aseptor. Tujuan penelitian ini adalah untuk memperoleh SPase rekombinan dari E. coli BL-21 StarTM dalam skala besar, memperoleh karakter enzim berdasarkan bobot molekul dan aktivitas, dan mengetahui aktivitas transglikosilasi terhadap asam askorbat dan asam benzoat. Berdasarkan hasil SDS PAGE, bobot molekul SPase rekombinan adalah antara 45 dan 66 kDa. Aktivitas SPase diukur dengan menggunakan sukrosa sebagai substrat dan produk akhir diukur sebagai NADPH pada 340 nm, dengan hasil aktivitas relatif 98,52% terhadap SPase standar. Aktivitas transglukosilasi menggunakan asam benzoat sebagai substrat menunjukkan produk pada KLT, sedangkan asam askorbat tidak menunjukkan terbentuknya produk pada KLT dan KCKT.

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Sucrosephosphorylase (SPase) is an enzyme that catalyzes glucosyl transfer reaction to the amount of acceptor molecules. The objective of this study was to obtain the recombinant SPase from E. coli BL-21 StarTM in a large scale, to characterize the recombinant SPase by its molecular weight and activity, and to determine the transglucosylation activity using ascorbic acid and benzoic acid as substrate.

SDS PAGE result showed that molecular weight of recombinant SPase between 45 and 66 kDa. SPase activity was measured by using sucrose as substrate, and the end product was measured as NADPH at 340 nm; this resulting relative activity 98.52% to standard SPase. Its transglucosylation activity using benzoic acid as substrate showed a product by performing TLC while as using ascorbic acid did not by performing TLC and HPLC."

"ABSTRAKSintesis senyawaan obat khiral secara organik konvensional pada umumnya produk yang dihasilkan merupakan campuran rasemat. Enzim lipase ekstraselular dari mikroba dapat mengkatalis reaksi kimia secara stereospesifik yang dapat menghasilkan enansiomer murni.Penelitian ini bertujuan untuk mempelajari kemampuan beberapa mikroorganisme lokal yaitu: Aspergillus niger UICC159, R.stolonifer UICC137, R.arrhizus UICC 2, R.oligosporus UICC 27, R.oryzae UICC 141 serta C.lipolyfica UICC Y-8, C.Tropicalis UICC Y-27 dan C.ufilis UICC Y-28 dalam menghasilkan lipase yang dapat meresolusi (R,S)-ibuprofen.Uji biotransformasi dari enzim lipase 4 isolat mikroorganisme terpilih yaitu kapang dan khamir Aspergillus niger UICC 159, Rhizopus stolonifer UCC 137 dan Rhizopus arrhizus UICC 2 dan Candida utilis UICC Y-28 dilakukan dengan penambahan (R,S)-ester metil ibuprofen berturut-turut setelah masa inkubasi 72, 72, 96 dan 40 jam.Hasil yang diperoleh menunjukkan bahwa Aspergillus niger UICC 159, Rhizopus stolonifer UICC 137, Rhizopus arrhizus UICC Y-2 dan Candida utilis UICC Y-28 memiliki %e.e : 89,59%, 72,61%, 99,63% dan 97,15 %.

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ABSTRACT

Optical Resolution of (R,S)-Ibuprofen by Enzymatic Biohydrolysis Screened From Several Local MicroorganismsThe conventional chemical synthesis of drug containing a chiral center generally yields an equal mixture of enantiomer. The extracellular microbial lipases are able to catalyse stereoselective chemical reaction for a mixture of rasemate, resolution enantiomer R(-) and S (+).

The aim of this study was to investigate the ability of several local microorganisms, such as : Aspergillus niger UICC 159, Rhizopus stolonifer UICC 137, Rhizopus arrhizus UICC 2 and Candida utilis UICC Y-28 to produce lipases applicable for resolution of (R,S)-Ibuprofen.

The biotransformation test of 4 isolates screened microorganism for lipase enzyme, i.e : fungi and yeast Aspergillus niger UICC159, Rhizopus stolonifer UICC 137, Rhizopus arrhizus UICC 2 and Candida utilis UICC Y-28 was done with subtrate addition incubation time 72, 72, 96 and 40 hour, respectively.

The result of enantiomeric excess from A. niger UICC 159, Rhizopus stolonifer UICC 137 , Rhizopus arrhizus UICC 2 and Candida utilis UICC Y-28 were obtained : 89,59 %, 72,61 % ,99,63 % and 97,15 %."

"[Latar belakang: Eksisi dini eskar luka bakar yang diikuti dengan autograft merupakan terapi utama pada luka bakar. Meskipun efektif, debridement dengan pembedahan secara teknis sulit dan mempunyai komplikasi. Debridement secara enzimatik menggunakan bromelain dapat mempreservasi epitelisasi dan tidak merusak jaringan sehat. Tujuan penelitian ini adalah menilai efek enzim bromelain produksi perusahaan lokal dikombinasi dengan hidrogel pada luka bakar dalam tikus. Metode penelitian: Tikus dibagi atas 3 kelompok yaitu kelompok 1 (luka bakar tanpa intervensi), kelompok 2 (luka bakar yang diberi gel), kelompok 3 (luka bakar yang diberi gel bromelain 10%). Kemudian masing-masing kelompok dibagi atas kelompok berdasarkan waktu 0,2,4,8,12,24 jam Reduksi eskar luka bakar difoto kemudian diukur menggunakan imageJ v 1.48®. Jaringan luka dibiopsi setelah binatang diterminasi dan diperiksa zone lisis, tipe dan derajat inflamasi. Hasil penelitian: Reduksi eskar luka bakar sedikit meningkat pada grup 1 dan 2 pada jam ke- 4 dan 24 (rata-rata 1.05% dan 2.2% pada jam ke-4, 3.52% and 4.13% pada jam ke-24). Gel bromelain sangat aktif merusak eskar luka bakar pada 4 jam pertama dan mencapai puncak pada jam ke-8 dan 12. Secara statistik terdapat perbedaan reduksi eskar dan zona lisis antara gel bromelain dan kontrol (p=0.000). Tipe inflamasi yang dominan pada semua grup adalah tipe campuran dan derajat inflamasi adalah sedang dan berat. Kesimpulan: Penetrasi gel bromelain 10% untuk mendegradasi eskar luka bakar optimal pada jam ke-8 dan ke 12, efektif untuk debridement eskar luka bakar dan tidak merusak jaringan sehat sekitarnya. Persentase PMN hampir sama pada semua grup dan secara statistik tidak terdapat perbedaan yang bermakna antara kelompok intervensi dan kontrol (p=0.47)., Background: Early excision burn eschar followed by autografting is a cornerstone of modern burn therapy. While effective, surgical debridement of the burned tissue is technically difficult and may cause considerable complications. Enzymatic debridement using bromelain can preserve the spontaneous epithelialisation potential and reduce the added injury to the traumatised tissue. The aim of the study was to assess the implication of bromelain enzyme that produce by local company combined with hydrogel on full-thickness skin burns of rats. Methods: Rats were divided in 3 group consist of group 1 (burn wound without intervention), group 2 (burn wound was treated with hydrogel), grup 3 (burn wound was treated with bromelain gel 10%). Each group was divided into subgroups time 0, 2, 4, 8, 12, 24 hours after intervention. The reduction of eschar surface area were measured by photographic documentation of the burns with ImageJ v1.48®. Histopathology preparations were made after terminated to measured lytic zone thickness, type and degree inflammation. Results : Burn eschar surface area reduction slightly increased in group 1 and 2 at 4 and 24 hours (mean 1.05% and 2.2% at 4 hours, 3.52% and 4.13% at 24 hours). Bromelain gel were most actively breaking down burn eschar during the first 4 hours. Peak of burn eschar reduction and lytic zone in the hours between 8 and 12 hours. There are statistically significant difference byrn eschar reduction and lytic zone between bromelain gel and control (p=0.000). The type of inflammation was a mixed inflammation type dominated and the degree of inflammation was moderate and severe in all group. Conclusion : Penetration of bromelain gel 10% optimally at 8 and 12 hours to degradation of the burn eschar and effective debride the burn eschar and has no apparent digestive effect on non-burned viable dermis and normal skin. Percentage PMNs almost similar in all groups and there is no statistically different between group of intervention and the control (p=0.47).]"

"Brazilin sebagai salah satu komponen aktif dalam kayu secang memiliki beragam kegunaan dan khasiat, yakni sebagai pewarna tekstil, pewarna alami makanan, dan media pengobatan herbal. Telah dikembangkan metode ekstraksi ramah lingkungan pada kayu secang sebagai alternatif penggunaan pelarut organic. Salah satunya adalah dengan penambahan enzim dalam proses ekstraksi yaitu dengan metode ekstraksi berbantu enzim (enzyme assisted-extraction/ EAE). Tujuan penelitian adalah meningkatkan kadar brazilin dan memperoleh kondisi optimum untuk ekstraksi brazilin dari kayu secang dengan enzim selulase kapang yang dibandingkan dengan metode refluks. Kandidat enzim selulase aktivitas tertinggi diproduksi dengan membandingkan hasil selulase kultivasi kapang Aspergillus niger UICC371, Trichoderma reesei IPBCC, dan campuran kedua isolat (1:1) dalam medium carboxymethyl cellulose cair. Serbuk kayu secang diekstraksi dengan enzim selulase hasil kultur cair dan selulase komersial masing-masing ditambahkan ke dalam pelarut akuabides pada variasi kondisi ekstraksi: konsentrasi enzim (2,0; 4,0; 6,0%); suhu ekstraksi (45, 50, 55℃); dan waktu esktraksi (1, 2, 3 jam). Desain variasi optimasi menggunakan respon permukaan (RSM)- BoxBehnken menghasilkan 15 kondisi perlakuan. Analisis brazilin menggunakan Kromatografi Cepat Kinerja Tinggi (KCKT) dengan fase gerak asetonitril : 0,3% asam asetat dalam air (14,5 : 85,5) selama 13 menit pada panjang gelombang 280 nm. Selulase kapang Aspergillus niger UICC371 aktivitas tertinggi (0,467 U/mL) dan selulase Aspergillus niger komersial dalam metode EAE menghasilkan kondisi optimum ekstraksi pada konsentrasi enzim 6,0% dan suhu 50℃. Penambahan selulase dalam ektraksi mampu meningkatkan kadar brazilin mencapai 5,014% dibandingkan metode refluks. Kondisi optimum berdasarkan anlisis RSM untuk konsentrasi enzim adalah 6,0%, suhu ekstraksi 50℃, dan waktu ekstraksi 1 jam.

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Brazilin has been known as one of active phytoconstituent from sappanwood that mainly present as textile colouring agent, food colouring, and herbal medicine purposes. Further extraction method in brazilin has been developed due to obtain maximum level of brazilin in sappanwood (Caesalpinia sappan L.) without organic solvent. Enzyme-assisted extraction (EAE) methods are currently one of the few types of methods in order to achieving that outcome. The following study aims to enhance brazilin level in sappanwood by achieving an EAE optimum condition by addition fungi cellulase compare to reflux extraction method. The cellulase candidates with highest activity are produced by compare the monoculture of fungi cellulase of Aspergillus niger UICC371, Trichoderma reesei IPBCC, and mixedculture (1:1) in carboxymethyl cellulose broth media. Sappanwood are extracted with fungi cellulase from submerged-fermentations production and commercial enzymes in aquabidest through multiple variation conditions: enzyme concentrations (2,0; 4,0; 6,0%); temperature (45, 50, 55℃); and time (1, 2, 3 hrs). The optimization are provided by response surface method-BoxBehnken design which form 15 different conditions. The brazilin level analysis carried out through High Performance Liquid Chromatography (HPLC) with asetonitril : 0,3% acetic acid in water (14,5 : 85,5) as eluents, for 13 mins in 280 nm wavelengths. The following study showed that cellulase from self-culture Aspergillus niger UICC371 are produced the highest activity (0,467 U/mL) and has been used in sappanwood-EAE method compare to commercial Aspergillus niger cellulase. The optimum condition of sappanwood-EAE methods were in 6,0% enzyme concentration and 50℃ temperature extraction which provide an increase in brazilin content up to 5,014% compare to reflux method. Response surface method for this EAE method were suggested in optimum condition by using 6,0% concentration enzyme at 50℃ for 1 hr time extraction."

"ABSTRACTProses ekstraksi hijau adalah metode yang digunakan untuk mendapatkan berbagai ekstrak tumbuhan dengan dampak minimal terhadap lingkungan. Ekstraksi hijau akan mengurangi konsumsi energi, memungkinkan penggunaan pelarut alternatif dan produk alami yang dapat diperbaharui, serta memastikan bahwa ekstrak yang dihasilkan aman dan berkualitas. Ultrasound-assisted enzymatic extraction UAEE adalah salah satu metode ekstraksi hijau. UAEE adalah metode ekstraksi yang mudah, efisien dan ramah lingkungan dan telah banyak digunakan untuk mengekstraksi berbagai jenis senyawa. Penggunaan enzim dalam metode ekstraksi ini akan mengkatalisis hidrolisis sitoderm dan glikoprotein, sehingga meningkatkan pelepasan zat bioaktif dengan mengganggu sel-sel tumbuhan. Kondisi operasi optimum yang digunakan dalam penelitian ini, yaitu konsentrasi enzim 70 mg/g, waktu hidrolisis enzimatik 2 jam dan konsentrasi etanol 50, akan menghasilkan yield ekstraksi maksimum sebesar 48,627. Crude extract daun keji beling diuji dengan menggunakan Gas Chromatography dan Mass Spectrometry GC-MS dan teridentifikasi senyawa yang memiliki aktivitas anti-hiperkolesterolemia, yaitu asam heksadekanoat, asam oktadekanoat dan skualen.

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ABSTRACTGreen extraction process is a method which is used to obtain various plant extracts with minimum impact on the environment. Green extraction will reduce energy consumption, allow use of alternative solvents and renewable natural products, and ensure a safe and high quality extract. Ultrasound assisted enzymatic extraction UAEE is one of green extraction method. UAEE is a mild, efficient and environmental friendly extraction method and it has been adopted for extracting various kinds of compounds. The use of enzyme will catalyze hydrolysis of the cytoderm and glycoproteins, therefore enhancing the release of bioactive substances by disrupting plant cells. The optimum extraction conditions with a maximum yield extract of 48.627 are as follows the concentration of ethanol is 50 and the amount of added enzyme is 70 mg g. Crude extract from keji beling leaves is tested using Gas Chromatography and Mass Spectrometry GC MS to identify components that have anti hypercholesterolemic activity, which are hexadecanoic acid, octadecanoic acid and demethyl squalene."

"Masalah kesehatan kulit merupakan hal yang menarik untuk diteliti lebih lanjut. Sudah terdapat banyak penelitian yang memusatkan kepada perkembangan atau penemuan solusi kesehatan kulit. Namun, penelitian mengenai produk kesehatan kulit yang bahan bakunya bersumber dari bakteri masih perlu untuk diteliti lebih lanjut, contohnya pemanfaatan postbiotik. Produk postbiotik yang saat ini banyak diteliti adalah lisat sel Bakteri Asam Laktat (BAL). Salah satu BAL yang memiliki berbagai potensi bagi kesehatan kulit adalah Streptococcus macedonicus MBF10-2, dimana telah terbukti menghasilkan senyawa seperti asam laktat yang bersifat sebagai pelembab, antimikroba dan meremajakan kulit, eksopolisakarida, dan peptida antimikroba/bakteriosin (lantibiotik macedocin dan macedovicin). Lisat sel Streptococcus macedonicus MBF10-2 diperoleh dengan pelisisan ultrasonikasi-enzimatis, dimana sebelumnya bakteri difermentasi dalam medium de Man, Rogosa dan Sharpe (MRS) Soy Peptone. Lisat yang diperoleh menghasilkan rendemen sebesar 3,98%, 5,51%, dan 5,02%, dengan rata-rata 5,625%. Cell Free Supernatant (CSF) bakteri juga diperoleh sebagai kontrol positif pada pengujian daya hambat. Pada pengujian daya hambat dengan metode mikrodilusi menghasilkan daya hambat lisat yang lebih lemah dibandingkan Cell Free Supernatant (CSF) bakteri, yang menunjukkan bahwa lisat S. macedonicus MBF10-2 masih membutuhkan penelitian lebih lanjut untuk melihat potensi lain selain aktivitas antimikrobanya.

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Skin health problems are interesting things for further research. There has been a lot of research that focuses on the development or discovery of skin health solutions. However, research on skin health products whose raw materials are sourced from bacteria still needs further investigation, for example postbiotic use. The postbiotic product that is currently being studied is lysate cell of Lactic Acid Bacteria (LAB). One BAL that has various potentials for skin health is Streptococcus macedonicus MBF10-2, which has been shown to produce compounds such as lactic acid which are moisturizing, antimicrobial and rejuvenating skin, exopolysaccharide, and antimicrobial/bacteriocin peptides (lantibiotic macedocin and macedovicin). Streptococcus macedonicus MBF10-2 cell lysates can be obtained by ultrasonication-enzymatic disruptiom, where previously bacteria were fermented in the medium de Man, Rogosa and Sharpe (MRS) Soy Peptone. The lysate yield that was calculated are 3.98%, 5.51%, and 5.02%, with an average of 5.625%. Bacterial Cell Free Supernatant (CSF) was also obtained as a positive control in inhibitory testing. In testing the inhibition by microdilution method concludes that the inhibition level of lysate is weaker than bacterial Cell Free Supernatant (CSF), which indicates that lysate of S. macedonicus MBF10-2 still needs further research to see other potentials besides its antimicrobial activity."

"Helicobacter pylori (H. pylori) is a gram negative and pleomorphic bacteria taht able to change its morphology according to environment...."

"This study investigates the enzymatic hydrolysis rate of Oil Palm (Elaeis guineensis) Trunk (OPT) sap in terms of the length of saccharification process with the aim to elevate sugar production. Emphasis was placed on the reaction time and addition of supplements such epsom salt (MgSO4) and alanine amino acid (C3H7NO2) to accelerate the efficiency of Saccharomyces cerevisiae containing the enzyme invertase. A whole oil palm trunk was divided into four different sections, upper, middle-1, middle-2 and bottom with separate experiments over 10 days enzymatic reaction period. The highest saccharification rate was shown as 13.47% on the tenth day. This result indicates that the increase in the saccharification rate was positively correlated with the length of hydrolysis. Moreover, the sample with nutrients achieved the highest sugar output, 17.91% on the fourth day of hydrolysis which was 4.44% higher than the hydrolysis rate of the sample without nutrients. In the presence of complex OPT sugars, together with other essential elements, epsom salt and alanine amino acid, S.cerevisiae achieved a higher hydrolysis metabolism to simple sugars as the cells strived to produce energy and regenerated the invertase. Moreover, the upper part of the OPT rendered the highest potential for sugar production with levels of 21.2% with supplements and 15.6% without. From this experimental analysis, a conventional saccharification method was optimized through the addition of nutrients and a prolonged (10 days) hydrolysis process which yielded an increase in sugar production."