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Surface response methodology for the optimization of lipase production under submerged fermentation by filamentous fungi.

ABSTRACT: A Plackett-Burman Factorial Design of 16 experiments was conducted to assess the influence of nine factors on the production of lipases by filamentous fungi. The factors investigated were bran type (used as the main carbon source), nitrogen source, nitrogen source concentration, inducer, inducer concentration, fungal strain (Aspergillus niger or Aspergillus flavus were selected as good lipase producers via submerged fermentation), pH and agitation. The concentration of the yeast extract and soybean oil and the pH had a significant effect (p<0.05) on lipase production and were consecutively studied through a Full Factorial Design 2(3), with the concentration of yeast extract and pH being significant (p<0.05). These variables were optimized using a central composite design, obtaining maximum lipolytic activities with the use of 45g/L of yeast extract and pH 7.15. The statistical model showed a 94.12% correlation with the experimental data.

SUBMITTER: Colla LM

PROVIDER: S-EPMC4874614 | biostudies-literature | 2016 Apr-Jun

REPOSITORIES: biostudies-literature

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Surface response methodology for the optimization of lipase production under submerged fermentation by filamentous fungi.

Colla Luciane Maria LM Primaz Andreiza Lazzarotto AL Benedetti Silvia S Loss Raquel Aparecida RA de Lima Marieli M Reinehr Christian Oliveira CO Bertolin Telma Elita TE Costa Jorge Alberto Vieira JA

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] 20160302 2

A Plackett-Burman Factorial Design of 16 experiments was conducted to assess the influence of nine factors on the production of lipases by filamentous fungi. The factors investigated were bran type (used as the main carbon source), nitrogen source, nitrogen source concentration, inducer, inducer concentration, fungal strain (Aspergillus niger or Aspergillus flavus were selected as good lipase producers via submerged fermentation), pH and agitation. The concentration of the yeast extract and soyb ...[more]

PMID: 26991270

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Project description:Although synthetic colorants are widely used in many industries due to their high stability at different conditions in industrial processes, evidence of its negative impact on health and the environment is undeniable. Filamentous fungi are well known for their use as alternative sources to produce natural pigments. However, an adequate comparison of the productivity parameters between the fermentation systems could be limited to their heterogeneous conditions. Even though Solid-State Fermentations (SSF) on natural substrates are widely used for pigments production, complex media, and non-controlled variables (T, pH, medium composition), these systems could not only hamper the finding of accurate productivity parameters, but also mathematical modeling and genomics-based optimization. In this context, the present study screened five pigment-producing fungi by comparing Submerged (SmF) and Surface Adhesion Fermentation [biofilm (BF) and Solid-State (SSF)] with defined media and controlled variables. For this purpose, we used the same defined media with sucrose as the carbon source for pigment production on SmF, BF, and SSF, and BF and SSF were carried out on inert supports. Five molecularly identified Penicillium and Talaromyces strains isolated from the Peruvian rainforest were selected for their ability to produce yellowish-orange colorants. Highest productivities were obtained from T. brunneus LMB-HP43 in SmF (0.18 AU/L/h) and SSF (0.17 AU/L/h), and P. mallochii LMB-HP37 in SSF (0.18 AU/L/h). Both strains also exhibited the highest yields (AU/g biomass) in the three fermentation systems, reaching values greater than 18-folds in SSF compared to the other strains. Conversely, T. wortmannii LMB-HP14 and P. maximae LMB-HP33 showed no ability to produce pigments in the SSF system. The performed experiments accurately compared the effect of the fermentation system on yield and productivity. From this, further genomics approaches can be considered for an extensive analysis of pigment synthesis pathways and a genomics-driven optimization in the best fermentation system.

Project description:BackgroundExtracellular production of fungal lipases especially the lipases obtained from the Aspergilli has gained immense interest in recent years due to its diverse biotechnological applications. In this study, we focused on determining the fermentation parameters required for the optimal lipase production.MethodsA total of 256 fungal isolates were obtained from oil seeds. From each genus, one isolate was selected to evaluate lipase production using phenol red and tributyrin plate assays. Lipase activity was estimated using the spectrophotometric pNPP hydrolysis assay. The highest lipase producer isolates were identified using 18S ribosomal RNA gene sequencing. The genetic variability was determined by random amplified polymorphic DNA (RAPD) analysis and the dendrogram was constructed using the unweighted pair group method with arithmetic averages method. The isolates were examined in a submerged fermentation culture (Smf) to measure the effect of temperature, pH, incubation time, carbon source, nitrogen source, inoculum volume, and lipid source on lipase production.ResultsEleven isolates belonging to the genus Aspergillus were analyzed for lipase production where they were found to be the highest lipase producers among various fungal genera. All the tested isolates were identified as A. niger using 18s rRNA sequencing. Genetic diversity was evaluated among all of the studied A. niger isolates using RAPD primers. The RAPD primers were used to amplify 285 loci, of which five were polymorphic (1.75%) and seven were monomorphic (2.45%). Thus, a high level of genetic diversity was observed among all isolates. The tributyrin test and the lipase activity assay identified five strains of A. niger as high lipase producers, and their optimal enzyme activities were 709.74, 532.54, 735.64, 794.62, and 787.69 U/ml. The optimal conditions for lipase production were as follows: 40 °C, pH 7.5, 1% fructose as the carbon source, 1% yeast extract as the nitrogen source, 2% palm oil, 2.5 × 107 spores/ml suspension, and 3 days of incubation.ConclusionsThe current study provides a comprehensive characterization of the optimal conditions, which are essential to enhance lipase production in five A. niger isolates.

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Surface response methodology for the optimization of lipase production under submerged fermentation by filamentous fungi.

Publications

Surface response methodology for the optimization of lipase production under submerged fermentation by filamentous fungi.

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