Judul: Collagenase of Bacillus licheniformis F11 from Palembang and application to produce bioactive collagen peptide
Abstrak: Collagenase is endopeptidase which can cleave helical collagen into small peptide fragments. The objective of the study was to select Bacillus licheniformis F11 which produced the best collagenase, purify collagenase dan characterize the crude and purified collagenase and apply collagenase from Indonesian Bacillus licheniformis in the production of bioactive collagen peptides which act as antioxidant, inhibitor of ACE (angiotensin I-converting enzyme) and anti cancer. The Bacillus licheniformis F11.1 and F11.4 were deleted gen from B. licheniformis F11, respectively. Collagenase from B. licheniformis was selected for application to hydrolyze fish skin collagen to produce collagen peptides. B. licheniformis 11.1 produced higher protease activity as compared to B. licheniformis F11.4 when grown in LB media. The optimum production time was 15 h for B. licheniiformis F11.1 with protease activity of 0.028 U/ml or 0.909 U/mg protein. For B. licheniformis F11.4 it was between 20-35 hours of fermentation, with enzyme activity of 0.005 U/ml or 0.157 U/mg protein. When collagen was added to the Modified LB media different responses were obtained. The activity of protease from B. licheniformis F11.1 decreased from 0.028 U/ml or 0.909 U/mg protein to 0.005 U/ml or 0.172 U/mg protein while that of B. licheniformis F11.4 increased from 0.005 U/ml or 0.157 U/mg protein to 0.017 U/ml or 0.546 U/mg protein. Based on result from extracellular collagenase production and spesificities towards substrate and finger print substrate, B. licheniformis F11.4 was selected for further study. Enzyme purification was done with ammonium sulphate 50% (w/v) precipitation, followed by DEAE Sephadex A-50 column chromatography. Precipitation with ammonium sulphate resulted in 5-fold purification with a yield of 10.1%. After purification with DEAE Sephadex A-50 column, the enzyme was purified 26.3-fold with a yield of 2.6%. The relative molecular weight was estimated to be 124 and 26 kDa. The optimum activity of both crude and purified collagenase (without and with Ca2+) were observed at 500C. The crude enzyme had pH optimum at 9.0, while the purified enzyme at 7.0. The crude collagenase activity was inhibited by FeCl2 (1mM) and CoCl2 (1 mM) while CuCl2 increased its activity. The purified collagenase activity was inhibited by CuCl2 (1 mM), FeCl2 (1 mM), MgCl2 (1 mM) and CoCl2 (1 mM). However, CaCl2 (10 mM), ZnCl2 (5 and 10 mM) and CuCl2 (5 and 10 mM) increased its activity. The radical scavenging activity (as measured by DPPH) of the collagen peptide hydrolysed with purified collagenase was higher (30.28%) than that of these hydrolysed with crude collagenase (23.50%). However, ferric ion reduction activity of the collagen peptide hydrolysed with crude collagenase (2.12) was higher than that of the collagen peptide hydrolysed with purified collagenase (0.78) and these were also higher than the 2.0 mM BHT antioxidant used as control. The ACE (Angiotensin I-Converting Enzyme) inhibitor activity of the collagen peptide hydrolysed with crude was higher (83%) as compared to that hydrolysed with purified collagenase (74%). The antiproliferation activity towards HeLa cervix cancer cells of the collagen peptide hydrolysed with purified collagenase (39.2%) was higher than that of the collagen peptide hydrolysed with crude (23.5%). The antiproliferation activity towards HCT-116 colon cancer
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Judul: Zimogram aktivitas kolagenase dari bacillus licheniformis F-11.1 dan F-11.4.
Abstrak: Bacillus licheniformis F-11.1 and F-11.4 are strains from B. lichniformis F-11 were isolated from shrimp shell waste in Palembang, Indonesia. They has poliglutamat acid mutation in the operon (PGA) and chitinase AB, so it doesn’t have chitinase activity, but high protease activity. Collagenase plays an important role in the softening of meat and the wound healing process. Precipitation enzyme can use ammonium sulfate to stabilize the enzyme, therebly reducing protein denaturation. Zimogram is an electrophoresis technique to determine the activity of an enzyme in situ and qualitative results of the gel electrophoresis. This research was conduct to analyze the activity Bacillus licheniformis F-11.1 and F-11.4 collagenase qualitatively using techniques zimogram. Analysis is performed on both isolates that grown in three growth media, ie LB + 5% collagen (LBC), ½ LB + 5% collagen (LBHC), and 5% collagen (CLG). Bacillus licheniformis F-11.1 and F-11.4 produce collagenase in the three growth media were added collagen as the substrate growth. Cell growth both isolates best on LBC media. Zimogram showed B. licheniformis F-11.1 and F-11.4 produce collagenase. It shown by the formation of clear zone on the gel due to degradation of collagen by collagenase. Isolates B. licheniformis F-11.1 produce the highest fraction of enzyme crude extract derived from LBC media is 32 fraction. Meanwhile isolates B. lichniformis F-11.4 produce the highest fraction of crude extract derived from LBHC media is 37 fraction
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Judul: Pola Pernikahan Masyarakat Batak Karo di Perantauan
Abstrak: Pernikahan adalah budaya yang tidak akan pernah berhenti, pernikahan akan terjadi secara berkelanjutan Dasarnya, karena pernikahan merupakan kebutuhan biologis manusia. Indonesia adalah negara yang kaya akan budaya dan etnis. Setiap suku di Indonesia mempunyai tata cara/adat istiadat dalam pelaksanaan pernikahan itu sendiri. Begitupun dengan suku Batak karo yang mempunyai sendiri tata cara pelaksanaan pernikahan. Seiring dengan perkembangan zaman Karo Banyak masyarakat yang merantau ke berbagai daerah di Indonesia, salah satunya Bogor berjalan-jalan. Masyarakat Karo yang merantau ke wilayah Bogor pada umumnya akan berusaha melakukan adaptasi ke negara tempat tinggal baru. Keadaan ini juga terjadi pada perkawinan Karo dalam pola adaptasi di Bogor. Namun adaptasi pernikahan masyarakat Karo di Bogor berupaya menjaga nilai-nilai budaya Karo dan sakralisme perkawinan. Mereka do Karo Dimodifikasi pada pola perkawinan, dengan tetap menjaga nilai budaya pernikahan.
Keyword: Weddings Karo, Adaptation marriage patterns, Attitudes