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Optimization of culture conditions by response surface methodology and unstructured kinetic modeling for bioactive metabolite production by Nocardiopsis litoralis VSM-8.

ABSTRACT: Response surface methodology-based central composite design on five variables incubation time, pH, temperature, sucrose concentration, and soya peptone concentration was employed for optimization of the production of bioactive compounds by Nocardiopsis litoralis strain VSM 8. The main quadratic effects and interactions of the five variables on the production of bioactive metabolites were investigated. A second-order polynomial model produced a satisfied fit for experimental data with regard to the production of the bioactive metabolites. Regression analysis showed that high R 2 values of all the five responses are significant and adjusted R 2 values showed good agreement with the experimental and predicted values. The present model was used to evaluate the direct interaction and quadratic effects to optimize the physico-chemical parameters for the production of bioactive metabolites that inhibit the pathogenic microorganisms measured in terms of zones of inhibition (responses). Mathematical kinetic model development and estimation of kinetic parameters also showed good approximation in terms of model fitting and regression analysis.

SUBMITTER: Managamuri U

PROVIDER: S-EPMC5055877 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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Optimization of culture conditions by response surface methodology and unstructured kinetic modeling for bioactive metabolite production by Nocardiopsis litoralis VSM-8.

Managamuri Ushakiranmayi U Vijayalakshmi Muvva M Poda Sudhakar S Ganduri V S Rama Krishna VS Babu R Satish RS

3 Biotech 20161008 2

Response surface methodology-based central composite design on five variables incubation time, pH, temperature, sucrose concentration, and soya peptone concentration was employed for optimization of the production of bioactive compounds by Nocardiopsis litoralis strain VSM 8. The main quadratic effects and interactions of the five variables on the production of bioactive metabolites were investigated. A second-order polynomial model produced a satisfied fit for experimental data with regard to t ...[more]

PMID: 28330291

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Project description:Deep sea sediment samples of Bay of Bengal (Visakhapatnam) have been analyzed for actinomycetes as an elite source to screen for the production of bioactive metabolites. The actinomycetes strain VSM-30 has an exciting bioactivity profile and was isolated during our systemic screening of marine actinomycetes. It was identified as Streptomyces sparsus based on morphological, physiological, biochemical, and molecular approaches. Response surface methodology regression analysis was carried out to fit the experimental data of each response by the second-order polynomial. The results have proven right interaction among process variables at optimized values of incubation time at 12 days, pH at 8, temperature at 30 °C, concentrations of starch at 1%, and tryptone at 1% and the data have been adequately fitted into the second-order polynomial models. Under these conditions, the responses (zones of inhibition) of plant pathogenic fungi Aspergillus niger, Aspergillus flavus, Fusarium oxysporum, Fusarium solani, and Penicillium citrinum were also matched with experimental and predicted results. Chemotypic analysis of ethyl acetate extract of the strain was done using LC-Q-TOF-MS revealed the presence of bioactive compounds including tryptophan dehydrobutyrine diketopiperazine, maculosin, 7-o-demethyl albocycline, albocycline M-2, and 7-o-demethoxy-7-oxo albocycline in a negative ion mode. The ethyl acetate extract of actinobacterium has been subjected to gas chromatography and mass spectroscopy (GC-MS) revealed the presence of diverse compounds such as dotriacontane, tetracosane 11-decyl-, diheptyl phthalate, 1-hexadecanesulfonyl chloride, L-alanyl-L-tryptophan, phthalic acid ethyl pentyl ester, 4-trifluoroacetoxyhexadecane, and 1H-imidazole 4,5-dihydro-2,4-dimethyl. Hence, the ethyl acetate extract of Streptomyces sparsus VSM-30 may have antibacterial, antifungal, and antioxidant activities due to the presence of secondary metabolites in ethyl acetate extract. The study also supports marine sediment samples of Bay of Bengal, a promising marine ecosystem remained to be explored for new bioactive compounds.

Project description:Modeling synthetic gene circuits to implement dynamic flux balancing is crucial in teaching and exploring metabolic engineering strategies to repartition metabolic precursors and construct efficient microbial cell factories. Microbial fitness and production rates are often complex phenotypes that are governed by highly non-linear, multivariable functions which are intrinsically linked through carbon metabolism. The solution of such dynamic system can be difficult for synthetic biologists to visualize or conceptualize. Recently, researchers (Santala et al., Metab. Eng. Comm., 2018) have implemented an arabinose based genetic switch to dynamically partition the central carbon flux between cell growth and product formation. The autonomous switch allowed dynamic shift from arabinose-associated cell growth to acetate-associated product (wax ester) formation. This system clearly demonstrates the effectiveness of using a genetic switch to decouple cell growth from product formation in a one-pot bioreactor to minimize operational cost. Coupled with Michaelis-Menten kinetics, and Luedeking-Piret equations, we were able to reconstruct and analyze this metabolic switch in silica and achieved graphical solutions that qualitatively match with the experimental data. By assessing physiologically-accessible parameter space, we observed a wide range of dynamic behavior and examined the different limiting cases. Graphical solutions for this dynamic system can be viewed simultaneously and resolved in real time via buttons on the graphical user interface (GUI). Metabolic bottlenecks in the system can be accurately predicted by varying the respective rate constants. The GUI serves as a diagnosis toolkit to troubleshoot genetic circuits design constraints and as an interactive workflow of using this arabinose based genetic switch to dynamically control carbon flux, which may provide a valuable computational toolbox for metabolic engineers and synthetic biologists to simulate and understand complex genetic-metabolic system.

Dataset Information

Optimization of culture conditions by response surface methodology and unstructured kinetic modeling for bioactive metabolite production by Nocardiopsis litoralis VSM-8.

Publications

Optimization of culture conditions by response surface methodology and unstructured kinetic modeling for bioactive metabolite production by Nocardiopsis litoralis VSM-8.

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