Dataset Information

Medium Optimization by Response Surface Methodology for Improved Cholesterol Oxidase Production by a Newly Isolated Streptomyces rochei NAM-19 Strain.

ABSTRACT: Cholesterol oxidase is an alcohol oxidoreductase flavoprotein with wide biotechnological applications. The current work describes the isolation of a potential cholesterol oxidase producing streptomycete from Egyptian soil. The isolated strain produced cholesterol oxidase in submerged culture using a medium containing glucose, yeast extract, malt extract, and CaCO3 with the addition of cholesterol as an inducer. The isolated strain was identified as Streptomyces rochei NAM-19 based on 16S rRNA sequencing and phylogeny. Optimization of cholesterol oxidase production has been carried out using response surface methodology. The Plackett-Burman design method was used to evaluate the significant components of the production medium followed by Box-Behnken experimental design to locate the true optimal concentrations, which are significantly affecting enzyme production. Results showed that the predicted enzyme response could be closely correlated with the experimentally obtained production. Furthermore, the applied optimization strategy increased volumetric enzyme production by 2.55 times (65.1?U/mL) the initial production obtained before medium optimization (25.5?U/mL).

SUBMITTER: Elsayed EA

PROVIDER: S-EPMC7150710 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Medium Optimization by Response Surface Methodology for Improved Cholesterol Oxidase Production by a Newly Isolated Streptomyces rochei NAM-19 Strain.

Elsayed Elsayed Ahmed EA Ahmed Abdelwahed Nayera N

BioMed research international 20200331

Cholesterol oxidase is an alcohol oxidoreductase flavoprotein with wide biotechnological applications. The current work describes the isolation of a potential cholesterol oxidase producing streptomycete from Egyptian soil. The isolated strain produced cholesterol oxidase in submerged culture using a medium containing glucose, yeast extract, malt extract, and CaCO3 with the addition of cholesterol as an inducer. The isolated strain was identified as Streptomyces rochei NAM-19 ba ...[more]

PMID: 32337225

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Project description:BackgroundCholesterol oxidases (CHOs) have attracted enormous attention because of their wide biotechnological potential. The present study explores the production of CHOs by Streptomyces sp. AN. Evaluation of culture conditions affecting enzyme production, medium optimization and released metabolite characteristics were also investigated.ResultsThe current work reports the isolation of 37 colonies (bacteria/actinobacteria) with different morphotypes from different soil/water samples. The isolate-coded AN was selected for its high potency for CHO production. Morphological characteristics and the obtained partial sequence of 16srRNA of AN showed 99.38% identity to Streptomyces sp. strain P12-37. Factors affecting CHO production were evaluated using Plackett-Burman (PB) and Box-Behnken (BB) statistical designs to find out the optimum level of the most effective variables, namely, pH, starch, NH4NO3 and FeSO4.7H2O with a predicted activity of 6.56 U/mL. According to this optimization, the following medium composition was considered to be optimum (g/L): cholesterol 1, starch 6, MgSO4.7H2O 0.1, CaCl2 0.01, FeSO4.7H2O 0.1, NH4NO3 23.97, yeast extract (YE) 0.2, K2HPO4 0.01, KH2PO4 0.1, NaCl 0.01, Tween 20 0.01, pH 6.36 and incubation temperature (30 °C) for 9 days. Spectophotometric analysis for released metabolites against cholesterol (standard) via Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) was carried out. FTIR spectrum showed the appearance of new absorption peaks at 1644 and 1725cm-1; this confirmed the presence of the Keto group (C=O) stretch bond. Besides, fermentation caused changes in thermal properties such as melting temperature peak (99.26; 148.77 °C), heat flow (- 8; - 3.6 Mw/mg), capacity (- 924.69; - 209.77 mJ) and heat enthalpy (- 385.29; 69.83 J/g) by comparison to the standard cholesterol as recognized through DSC thermogram. These changes are attributed to the action of the CHO enzyme and the release of keto derivatives of cholesterol with different properties.ConclusionStreptomyces sp. AN was endowed with the capability to produce CHO. Enzyme maximization was followed using a statistical experimental approach, leading to a 2.6-fold increase in the overall activity compared to the basal condition. CHO catalyzed the oxidation of cholesterol; this was verified by the appearance of a new keto group (C=O) peak at 1644 and 1725 cm-1 observed by FTIR spectroscopic analysis. Also, DSC thermogram demonstrates the alteration of cholesterol triggered by CHO.

Dataset Information

Medium Optimization by Response Surface Methodology for Improved Cholesterol Oxidase Production by a Newly Isolated Streptomyces rochei NAM-19 Strain.

Publications

Medium Optimization by Response Surface Methodology for Improved Cholesterol Oxidase Production by a Newly Isolated <i>Streptomyces rochei</i> NAM-19 Strain.

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