Comparison of protein content extracted from green algae Kirchneriella sp. Schmidle lyophilized biomass using different determination methods
Author | Affiliation |
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Koreivienė, Judita | Gamtos tyrimų centras |
Date |
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2020-10-23 |
History of Pharmacy Section. Abstracts. Students section.
ISBN 978-9955-15-669-7.
Bibliogr.: p. 19
Introduction: Micro-algal protein has potential for animal feed or human consumption, recombinant protein technology and as a valuable by-product of biofuel production. Techniques that can be used to measure protein content rapidly in lyophilized material include methods of elemental analysis and Kjeldahl to measure N-content. Micro-algae often contains substantial levels of free amino acids and other N-organic compounds such as chlorophyll. Dye-based procedures such as Bradford and Lowry largely overcome these problems and are more appropriate when applied to a wide variety of strains such as high-throughput screening procedures[1]. Materials and methods: Materials – lyophilized biomass of green algae Kirchneriella sp. Schmidle, was received from the Nature Research Centre, Laboratory of Algology and Microbial Ecology. Methods: protein extraction was carried out using 0.2 M NaOH solution; protein precipitation using mixture of 13.3 % trichloroacetic acid (TCA) and 0.2% β-mercaptoethanol (β-ME) in acetone; protein pellets were washed with 100% acetone; proteins were resuspended in phosphate buffer solution (PBS) pH 7.4. The protein content was determined by spectrophotometric microplate Bradford and Lowry methods. Concentrations of isolated proteins were based on the calibrations curve of bovine serum albumin (BSA). Statistical analysis based on Microsoft Excel [2-3]. Results: It was determined by Lowry microplate method that in 1.0 g of lyophilized biomass of Kirchneriella sp. Schmidle is 973.75±32.36 μg of proteins. 2.72±0.29 μg of proteins was determined by Bradford Well Plate assay. Conclusions: The obtained results show that the protein content determined in lyophilized biomass of Kirchneriella sp. Schmidle by the Bradford method is significantly lower than by the Lowry method. Lower concentrations of protein using Bradford’s method may be related to the binding of the dye Brilliant Blue-G250. The Folin reagent used in the Lowry assay interact