Investigation of low-temperature lipase production and enzymatic properties of Aspergillus Niger (original) (raw)
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International journal of pharma and bio sciences, 2013
Lipase production in Aspergillus niger was tested using both submerged fermentation (SmF) and solid-state fermentation (SSF) on a mineral culture medium and wheat bran, respectively.Culture media was optimized in both SmF and SSF. We found 1.46 IU/mL was the optimum activity of lipase in case of submerged fermentation, when medium containing 2% glucose and 2% olive oil under the conditions of 1 vvm and 450 m -1. However, 9.14 IU/g of dry solid substrate equivalent to 4.8 IU/mL of lipase activity was reached using solid-state fermentation process with a medium containing 0.75 % of ammonium sulphate and 0.34 % of urea. The optimum pH and temperature for enzymatic activity were pH=6 and 40 °C, respectively. When we used neutral and midly acid media, temperature ranges between 20 to 30⁰C for 24 hour, the enzyme showed 80% of its initial activity.
International Journal of Food Science & Technology, 2014
Lipase from Aspergillus sp. obtained by solid-state fermentation (SSF) on wheat bran (LWB), soybean bran (LSB) and soybean bran combined with sugarcane bagasse (LSBBC) were 67.5, 58 and 57.3 U of crude lipase per gram substrate, respectively. The optimum pH of activity and stability of the LWB was between 8 and 9, and the optimum temperature of activity and stability was 50°C and up to 60°C, respectively. The LSB and LSBBC showed two peaks of optimum pH (4 and 6) and optimal values of temperature and stability at 50°C. The LSB was stable in the pH range of 6-7, while LSBBC in the range of pH 4-7. All the enzymes show activities on p-nitrophenyl esters (butyrate, laurate and palmitate). LWB stood out either on the hydrolysis of sunflower oil, presenting 66.1% of the activity over commercial lipase and on the esterification of oleic acid and ethanol, surpassing the activities of the commercial lipases studied. The thin layer chromatography showed that LWB and LSB have produced ethyl esters from corn oil, while LWB produced it from sunflower oil.
Production of acidic lipase by Aspergillus niger in solid state fermentation
Among the various fungal strains screened for lipase production, Rhizopus arrhizus NCIM 877, 878, 879 and Aspergillus niger NCIM 1207 produced significant quantities of enzyme when grown in synthetic oil based (SOB) medium under submerged conditions. Rhizopus strains showed major intracellular activity while A. niger NCIM 1207 produced mainly extracellular activity. Lipase production in A. niger NCIM 1207 was studied using both submerged fermentation (SmF) and solid state fermentation (SSF). De novo biosynthesis of lipase occurred only in the presence of lipid substrate and was completely repressed by glucose. The highest yields of enzyme were obtained in SSF using wheat bran as solid substrate in combination with olive oil as lipid substrate. Maximum lipase activity (630 IU/g dry solid substrate) was recovered when fermented wheat bran was extracted with NaCl (1%) supplemented with Triton X-100 (0.5%). The pH and temperature optima for lipase were 2.5 and 45 8C, respectively. The enzyme also exhibited high activity (75%) at extremely acidic pH of 1.5. Lipase activity (63%) was retained when enzyme was incubated in a buffer of 2.5 for 24 h at room temperature. The enzyme retained 63% of its original activity on incubation at 70 8C for 5 h. This organism, being GRAS cleared, can be used for large-scale production of enzyme for commercial purpose.
2004
Of the 34 fungal species, isolated from a number of oily substrates, 9 exhibited lipase activity. AU 15, identified as Aspergillus sp., was found to be excellent lipase producer in submerged fermentation and was selected for solid-state fermentation (SSF). Among substrates like oil cakes of coconut, groundnut and sesame, wheat rawa, bombay rawa and soya beans (crushed), wheat rawa showed the highest lipase activity. The maximum enzyme yield (1934 U/g) was obtained with basal medium containing wheat rawa, olive oil and corn steep liquor, at 80% moisture content, pH 7.0 and 96 hrs incubation.
Indonesian Journal of Biotechnology, 2017
Jatropha curcas seed cake contains a high amount of protein, and consequently has very high potentialas a medium for lipase production. The objective of this research was to characterize lipase from Aspergillusniger 6516, which was produced by solid-state fermentation on Jatropha curcas seed cake as the medium. The effects of pH and temperature on enzyme activity were evaluated, along with substrate specifcity and enzyme stability. Fermentation was performed at a water concentration of 63% and temperature of 30 °C for 7 days. The results showed that the optimum pH and temperature for Aspergillus niger 6516 lipase activities were 8.0 and 40 °C, respectively. The lipase had the substrate specifcity to hydrolyze long-chain fatty acids and was stable in polar organic solvents. The lipase had a molecular weight, Km and vmax about 19 kDa, 0.27 µmol/ml, and 52.63 µmol/ml/min, respectively. The results also suggested that the produced lipase from Aspergillus niger 6516 was an alkaline lipas...
Biomedicine and Biotechnology, 2014
An enzyme with various commercial purpose, lipase is a carboxy esterase enzyme with many uses in different industries. Multiple isolates of Aspergillus niger were isolated from oil contaminated soil samples and screened for lipase producing ability on tributyrin medium. The isolate showing maximum activity was identified and subjected to growth parameters optimization in attempt to increase the enzyme producing ability of the isolate in larger scale. Different media with varying composition were examined for best lipase production. The activities of the lipase produced by the fungus at various pH were assessed. The enzyme activity was determined by the titration method. Maximum lipase activity of 2.4 U/ml was achieved with organic nitrogen rich media (designated as PM II) at pH 7 on the sixth day of culture. The lipase production was scaled up on a pilot scale in a 5 Liter fermenter maintaining growth parameters of pH 7, temperature at 28°C, stirrer rate at 120 rpm, airflow rate at 30 L/hr, O2 saturation 50% and pressure 0.05 MPa. The crude enzyme was extracted for further assays. Optimization of the parameters can improve the productivity as well as the quality of the enzyme produced.
Production and optimization of lipase using Aspergillus niger MTCC 872 by solid-state fermentation
Bulletin of the National Research Centre
Background: Lipases are serine hydrolases that degrade triglycerides, an attribute that treasures wide applications in biodiesel production, detergent, chemical industries, etc. The most sought after the application is in the high quality and economical production of biodiesel under mild reaction conditions and simplified product separation. For the said application, fungal lipases are ideal catalysts that could effectively catalyze esterification and transesterification reactions with their specific ability to release fatty acids from 1, 3 positions of acylglycerols. Results: In the present work, to facilitate bulk synthesis, lipase production using Aspergillus niger MTCC 872 was studied by solid-state fermentation (SSF). The chosen fungal strain was evaluated for lipase production using a mixture of agroindustrial substrates viz. rice husk, cottonseed cake, and red gram husk in various combinations at flask level. Tri-substrate mixture (rice husk, cottonseed cake, and red gram husk) combined in the ratio of 2:1:1 has shown the maximum lipase activity 28.19 U/gds at optimum cultivation conditions of temperature 40°C, moisture content 75% (v/w), pH 6.0 and initial spore concentration of 5.4 million spores per mL. Further studies were performed for scale-up of lipase from flask level to lab scale using tray fermenter. Lipase activity was found to be 24.38 U/gds and 21.62 U/gds for 100 g and 1000 g substrate respectively. Conclusion: This is the first report on the production of lipase from Aspergillus niger MTCC 872 using tri-substrate mixture of rice husk (RH), cottonseed cake (CSC), and red gram husk (RGH). Moreover, comparison between individual, binary, and tri-substrate mixture was carried out for which the highest lipase activity was observed for tri-substrate mixture. In addition, comparable results were found when scale-up was performed using tray fermenter. Thus, the current work signifies usage of agro-industrial residues as substrates for enzyme production by solid-state fermentation process as an effective alternative to submerged fermentation for industrial applications.
Energy Reports, 2020
Lipase is one of the most used industrial enzymes, which can be produced by Aspergillus niger. Large scale production of Aspergillus niger lipase is more profitable by using solid state fermentation method and utilizing agro-industrial waste as the substrate. Optimization of Aspergillus niger lipase production has been performed in this study. Optimization was performed on solid state fermentation of rice bran and Jathropa seed cake for 5 days with variations of inducer and extractant. Fermentation cake produced was extracted, filtered using muslin cloth, and centrifuged. The supernatant was spray-dried and assayed using olive oil hydrolysis titrimetry. Inducer optimization results showed that 1% of olive oil was the best inducer, yielding dry lipase extract with highest activity unit (176 U/ml enzyme). Extractant optimization results showed that 1% of NaCl-0.5% of Tween 80 was the best extractant, yielding dry lipase extract with highest activity unit (282 U/ml enzyme). c
Characterization of Lipase from Wild (LPF-5) and Mutant (HN1) Strain of Aspergillus niger
International Journal of Current Microbiology and Applied Sciences, 2016
Lipase is one of the most imperative industrial enzymes and has a variety of applications in various industries. In the present study, lipases obtained by submerged cultivation of wild (LPF-5) and mutant (HN1) strain of A. niger were used and compared for characterization study. Activity and stability of partially purified lipase was determined under different pH, temperature, organic solvents and metal ions. The lipase showed highest activity and stability at pH 7.0 and temperature 35 °C for LPF-5 and 30 °C for HN1 strain. The lipases retained high activity over ranges of temperature (25-50 ºC) and pH (4.0-7.5). Lipase was enhanced by methanol and acetone, while slightly inhibited by butanol (10% v/v). Ca 2+ appeared to be the excellent inducer of lipase activity. Lipase also showed stability in presence of the other metal ions (Na + , Ba 2+ , Mg 2+ , Cu 2+ , Fe 2+ and Mn 2+). The lipase of wild and mutant strain retained 10.58% and 14.60% of its activity when pre-incubated with Hg 2+ , indicating the inhibitory effect of Hg 2+ on catalytic activity of lipase. K e y w o r d s A. niger, mutant strain (HN1), wild strain (LPF-5), temperature, pH, metal ions, methanol, characterization.
Extracellular lipase produced by Aspergillus japonicus was tested for its activity on sesame, groundnut and sunflower oil substrates. The enzyme showed significant activity in the pH range of 6-8 and a temperature range of 300C-400C but with sunflower oil, the activity is optimum at 500C. This is indicative of the substrate playing a role in the optimization of physico-chemical characteristics affecting the enzyme activity. Of the eleven metal ions tested, only Mg2+ (2mM) enhanced the enzyme activity while others, inhibited. EDTA significantly (P <0.05) enhanced the enzyme activity suggesting that metal ions do not in general affect the lipase isolated from A. japonicus. Organic solvents and acids tested showed significant (P< 0.05) enhancement of the lipase activity at higher concentrations, presumably because of their influence on the interfacial area. Km and Vmax values of the partially purified lipase determined from Lineweaver Burk and Eadie-Hofstee plots were 63.09 (mM/L), 5.33 (mM/L/min) and 71.76 (mM/L), 5.25 (mM/L/min) respectively