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PEMURNIAN PARSIAL DAN KARAKTERISASI AMILASE DARI BAKTERI LAUT ARTHROBACTER ARILAITENSIS LBF-003 Rahmasari, Dianti; Wijanarka, Wijanarka; Pujiyanto, Sri; Rahmani, Nanik; Yopi, Yopi
JURNAL BIOLOGI INDONESIA Vol 12, No 1 (2016): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v12i1.2323

Abstract

Starch is an abundant carbon source in nature, and ?-amylase (1, 4-?-D-glucanohydrolase; EC 3.2.1.1), which hydrolyzes ?-1, 4-glucosidic linkage in starch-related molecules. Microbe ?-amylase production is a hydrolytic enzyme and one ofinterest in its microbial production has increased dramatically due to its wide spread use in food, textile, baking anddetergent industries in recent years. Here we report ?-amylase from marine bacterium which was purified andcharacterized, as well as analyzed its hydrolysis product on starch. The enzyme of Arthrobacter arilaitensis partiallypurified by acetone precipitation with 90% and ion exchange chromatography produced specific activity 0.25 U/mg and0.38 U/mg, and it?s purity rate increased until 1.14 fold compared with former crude extract. Purifed extracelluler amilasehad an optimum activity at temperature 50°C and pH 9.0. An apparent molecular mass was between 50-75 kDa, asestimated by zimogram electrophoresis. Hydrolysis products of this enzyme on starch were maltose, maltotriose andmaltoheptaose.Keywords: alfa amylase, marine bacterium, Arthrobacter arilaitensis, purification, charaterization
Enzymatic Hydrolysis of Mannan from Konjac (Amorphophallus sp.) Using Mannanase from Streptomyces lipmanii to Produce Manno-oligosaccharides Sasongko, Ashadi; Yopi,; Rahmani, Nanik; Lisdiyanti, Puspita; Saepudin, Endang
Makara Journal of Science
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Mannan is an abundant polysaccharide that can be found in konjac (Amorphophallus sp.). Mannan can be enzymatically hydrolyzed using mannanase to produce manno-oligosaccharides which can be used as a prebiotic. The aims of this research are to determine the production time of mannanase from Streptomyces lipmanii, perform enzyme characterization, optimize the hydrolysis time, and characterize the hydrolysis product. A qualitative assay using the indicator Congo red showed that S. lipmanii generated a clear zone, indicating that S. lipmanii produced mannanase in konjac medium and possessed mannanolytic activity. Enzyme activity was determined through reducing sugar measurement using the dinitrosalycylic acid method, and optimum enzyme production was achieved at the second day of culture. Characterization of the enzyme showed that hydrolysis was optimum at pH 7 and at a temperature of 50 oC. The reducing sugar content was increased by an increasing the hydrolysis time, and reached an optimum time at 2 h. The degree of polymerization value of three was achieved after 2 h hydrolysis of mannan from konjac, indicating the formation of oligosaccharides. Analysis by thin layer chromatography using butanol, acetic acid, and water in a ratio of 2:1:1 as eluent showed the presence of compounds with a retention time between those of mannose and mannotetrose. Confirmation was also performed by HPLC, based on the retention time.