Molecular, physiological, and biochemical characterization of extracellular lipase production by Aspergillus niger using submerged fermentation (original) (raw)
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Different lipid rich products were used to obtain oil degrading fungal isolates. The isolates were codified for referral to our culture bank and compared for their lipolytic potential. Amongst the isolates, MBL-1412 isolated from the cooked "sliced cicer arietinum" (Channa Daal) was found to be a potent hyper-producer and was optimized for lipase production under solid state fermentation. Initial systematic treatment based upon micrometric data and consultation with the standard monographs and fungus ended up with its identification as Aspergillus sp. The identification confirmed that the fungus belongs to genus Aspergillus, by DNA barcoding marker like 18S rRNA gene sequence.Later, the sequence was registered with accession no. KM924434 in the public nucleotide library (genbank) of NCBI. Fungal culture was maintained on 2% potato dextrose agar (PDA) during the study. Diverse substrates of agricultural byproducts under varied incubation temperature, time interval, inoculum level and different pH of diluent were used as parameters of optimization for hyperproduction of lipases. Different carbon and nitrogen sources as additives of culture medium were applied for enhancement of lipase production. Almond meal (10g) with inoculum level @1.5 mL after 48 h of time course at 50ºC and 6 pH were selected to be the best eco-cultural conditions for optimal lipases production by Aspergillus sp. MBL-1412. Supplementary additives of nitrogen and carbon sources to the basal substrate improved lipases production appreciably. Ammonium chloride (1%) as inorganic nitrogen source, nutrient broth (0.8%) as organic nitrogen source and starch (0.8%) as carbon source were found as best media additives for enhanced extracellular lipases yield.
2018
Lipases are the enzymes of choice for applications in a large number of industries. Present study describes the isolation of potent lipolytic fungal strains and their subsequent production by using shake flasks fermentation technique. The selected isolate was identified as Aspergillus terreus, based on morphological features and 18s rRNA sequencing. Seven different culture media were analyzed for the extracellular lipase production employing A. terreus as the production organism and it was found that M6 gave maximum lipase production i.e., 5.0 U/mL/min in the medium containing (% w/v) MgSO4, 0.05; NaNO3, 0.05; KCl, 0.05; KH2PO4, 0.2; olive oil, 1.0; pH 6.0. Maximum production of lipase (7.66 U/mL/min) was found at a medium pH of 6.0 and an incubation temperature of 30C after 72 hrs of incubation period. An increase in the lipase production (7.99 U/mL/min) was observed when the production medium was provided with 1.0% peptone, 2.0% glucose and 1.0% olive oil. Ammonium sulphate precip...
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.
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.
Journal of Basic Microbiology, 2010
Under the current assay conditions, lipase production in mineral medium was only detected in the presence of vegetable oils, reaching the highest specific activity with olive oil. In this way, effect of different environmental conditions on fungal morphology and olive oil-induced extracellular lipases production from Aspergillus niger MYA 135 was studied. It was observed that addition of 1.0 g l -1 FeCl 3 to the medium encouraged filamentous growth and increased the specific activity 6.6 fold after 4 days of incubation compared to the control. However, major novelty of this study was the satisfactory production of an acidic lipase at initial pH 3 of the culture medium (1.74 ± 0.06 mU μg -1 ), since its potencial applications in food and pharmaceutical industry are highly promising.
International Journal of Scientific Research in Environmental Sciences, 2016
Twelve fungal isolates belonging to different genera were quantitatively screened for extracellular lipase production in SmF (Submerged fermentation). Isolate LPF-5 demonstrated higher lipase activity and it was identified as Aspergillus niger based on morphology of its Petri plate culture, microscopic study of its slide culture and with the help of relevant literatures. Further an effort was made to increase the production of extracellular lipase by subjecting the most potent lipolytic fungal strain A. niger LPF-5 to the strain improvement by induced mutagenesis using UV radiations and nitrous acid. Among the all tested mutagens, nitrous acid was found to be the best mutagen (incubation time of 15 minutes) for inducing the favorable mutation in fungal strain A. niger LPF-5 which enhanced the lipase activity up to 30% (105.19 ± 0.91 U mL-1 min-1) when compared to lipase activity (81.00 ± 0.30 U mL-1 min-1) of wild strain while the lipase activity of the best UV mutant (A. niger UV3), obtained after an incubation of 6 minutes was 94.30 ± 0.54 U mL-1 min-1 , which was 16.41% higher as compared to wild strain of A. niger. These results indicate that UV light and nitrous acid both were effective mutagens but nitrous acid was more potent mutagen than UV light.
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 and Partial Characterisation of Extracellular Lipase From Aspergillus Niger
Biotechnology …, 1996
The production and certain kinetic characteristics of extracellular lipase from Asperbai/!us niger were investigated. It was possible to substantially enhance the activity of excreted lipase by optimising the interaction between carbon and nitrogen sources applying a two-parameter complete experimental design and response surface analysis. The enzyme was partially purified and a number of kinetic characteristics such as optimum pH and temperature, thermal and pH stability and K, were determined and discussed.'The elevated levels of lipase activity (40.5 U/ml) found in this work competed favourably with most of those reported for lipase hyperproducing fungi.
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.
Journal of Basic Microbiology, 2010
Extracellular lipase production by Aspergillus sp. (RBD-01) was monitored by modulating pH of the growth medium, ambient temperature for growth, source of nitrogen and percentage of carbon (virgin cottonseed oil). This strain was observed to be viable and produces lipase even up to 50% oil as a main carbon source. Maximum lipase activity of 21.8 U/ml was obtained with 50% (v/v) oil acting as the main carbon source and peptone (0.5% w/v) as nitrogen source. The optimum pH and temperature for enzymatic activity were observed to be 7.5 and 35 °C, respectively. The observations are of significance due to limited reports on use of 50% of oil as the main carbon source while obtaining significant lipase activity of 21.8 U/ml.