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Lipase Production by Aspergillus niger using Solid State Fermentation with Candlenut Seed as Substrate
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| Author:
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DEBORAH BASA, NOPHERINA ARUAN, ROGA FLORIDA KEMBAREN, MERRY MERYAM MARTGRITA
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| Abstract:
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Lipase is the enzyme that hydrolyzes insoluble long chain triglycerides into monoglycerides, diglycerides, glycerol, and fatty acids. Lipase is widely used in industries due to its ability to do specific chemical transformation. Aspergillus niger is frequently used to produce lipase, due to its fast grow characteristic, and can produce thermostable and extracellular lipase. A. niger can produce lipase when cultured in several kinds of substrate. The objective of this research is to produce lipase by A. niger using solid state fermentation (SSF) and candlenut seed as substrate, that has never been observed before. Candlenut seed was used due to its high lipid content (50-60% w/w). Extracellular lipase was separated from the growth media by centrifugation. Lipase was purified by ammonium sulfate precipitation (40%-80%) and then dialyzed in 1 mM Tris-HCl buffer pH 8.0. Lipase activity was determined spectrophotometrically using p-nitrophenyl acetate (p-NPA) as substrate. Lipase with the highest activity (4.71 U/ml) was obtained from the culture with 15 g of candlenut seed substrate and partially purified by 80% ammonium sulfate. Optimum condition of lipase was characterized using Central Composite Design (CCD) method in pH range of 4.0-8.0 and temperature range of 30°C-50°C. The result showed that optimum condition of lipase is in pH 6.03 and temperature of 41oC with activity of 49.21 U/ml. The optimum condition was used for hydrolyzing candlenut oil by lipase. Gas chromatography analysis of the hydrolysis products showed that lipase is able to hydrolyze candlenut oil into fatty acids. It can be concluded that candlenut seed is a potential substrate for producing lipase from A. niger by solid state fermentation.
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| Keyword:
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Aspergillus Niger, Candlenut Seed, Central Composite Design, Lipase, Solid State Fermentation
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2020.12.02.366
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