scholarly journals Response surface methodology to optimize inulinase production by a newly isolated Penicillium amphipolaria strain by solid-state fermentation of Saccharum arundinaceum

2020 ◽  
Author(s):  
Deblina Das ◽  
Ramananda M Bhat ◽  
Raja Selvaraj

Abstract In the present investigation, a new fungal inulinase producer Penicillium amphipolaria KAS 2555 has been isolated from the soil of dead mangroves litter area, followed by the inulinase production and optimization by solid-state fermentation using a low-cost substrate – hardy sugarcane (Saccharum arundinaceum). While screening, only Penicillium amphipolaria KAS 2555 showed the hydrolysis zone on the plate containing inulin media. The exoinulolytic nature of inulinase and its form of action was confirmed by thin-layer chromatography (TLC). After 96 h of the fermentation period, an activity of 2.45 U/gds was obtained. The I/S ratio of 0.59 proved that the enzyme is inulolytic in nature. Media optimization was performed to obtain a regression model using Central Composite Design (CCD). For optimization, five significant media components viz., inulin, (NH4)2SO4, K2HPO4, KH2PO4 and, NaCl were used. A 3.10-fold increase in activity of inulinase (7.59 U/gds) was obtained under the optimal settings of (g/gds) inulin- 0.1, (NH4)2SO4- 0.002, K2HPO4- 0.1, KH2PO4- 0.02 and NaCl- 0.02.

2021 ◽  
Author(s):  
Daniel Otieno Ojwang ◽  
Mulaa F. Jakim ◽  
George Obiero ◽  
Jacob Midiwo

Abstract Mushrooms are a rich source of high value compounds. Efficient enzymatic degradation of mushroom cell-wall matrix is therefore critical in the recovery of cellular components in high yields. In the present study, the effect of reaction variables on mushroom saccharification using crude enzymatic extract was evaluated and optimized using the central composite design based on the response surface methodology. The crude extract displayed CMCase, Fpase and xylanase activities of 1.23Umg− 1 protein, 0.95Umg− 1 protein and 1.52 Umg− 1 protein. The model was validated by the analysis of variance with a coefficient of determination of 0.866 and F test value of 7.39, making the model valid at the 95 % confidence limit. The model achieved glucose yield of 1.490mg/mL at pH 6.5, temperature 50oC, enzyme load of 5% (v/v) and reaction time of 12h. The experiment using optimal model conditions yielded 1.582 mg/mL glucose which is 1.1 folds higher than the predicted model value. This study demonstrated potential of crude extract from solid-state fermentation of low-cost pineapple peels in mushroom processing.


2020 ◽  
Vol 16 (3) ◽  
pp. 246-253
Author(s):  
Marcin Gackowski ◽  
Marcin Koba ◽  
Stefan Kruszewski

Background: Spectrophotometry and thin layer chromatography have been commonly applied in pharmaceutical analysis for many years due to low cost, simplicity and short time of execution. Moreover, the latest modifications including automation of those methods have made them very effective and easy to perform, therefore, the new UV- and derivative spectrophotometry as well as high performance thin layer chromatography UV-densitometric (HPTLC) methods for the routine estimation of amrinone and milrinone in pharmaceutical formulation have been developed and compared in this work since European Pharmacopoeia 9.0 has yet incorporated in an analytical monograph a method for quantification of those compounds. Methods: For the first method the best conditions for quantification were achieved by measuring the lengths between two extrema (peak-to-peak amplitudes) 252 and 277 nm in UV spectra of standard solutions of amrinone and a signal at 288 nm of the first derivative spectra of standard solutions of milrinone. The linearity between D252-277 signal and concentration of amironone and 1D288 signal of milrinone in the same range of 5.0-25.0 μg ml/ml in DMSO:methanol (1:3 v/v) solutions presents the square correlation coefficient (r2) of 0,9997 and 0.9991, respectively. The second method was founded on HPTLC on silica plates, 1,4-dioxane:hexane (100:1.5) as a mobile phase and densitometric scanning at 252 nm for amrinone and at 271 nm for milrinone. Results: The assays were linear over the concentration range of 0,25-5.0 μg per spot (r2=0,9959) and 0,25-10.0 μg per spot (r2=0,9970) for amrinone and milrinone, respectively. The mean recoveries percentage were 99.81 and 100,34 for amrinone as well as 99,58 and 99.46 for milrinone, obtained with spectrophotometry and HPTLC, respectively. Conclusion: The comparison between two elaborated methods leads to the conclusion that UV and derivative spectrophotometry is more precise and gives better recovery, and that is why it should be applied for routine estimation of amrinone and milrinone in bulk drug, pharmaceutical forms and for therapeutic monitoring of the drug.


2017 ◽  
Vol 118 ◽  
pp. 19-26 ◽  
Author(s):  
Mohd Anis Ganaie ◽  
Hemant Soni ◽  
Gowhar Ahmad Naikoo ◽  
Layana Taynara Santos Oliveira ◽  
Hemant Kumar Rawat ◽  
...  

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