A comparative study on optimisation of protein extraction methods for Saccharomonospora azurea

Fodor István; Valasek Andrea; Urbán Péter; Kovács Márk; Fekete Csaba; Kerepesi Ildikó: A comparative study on optimisation of protein extraction methods for Saccharomonospora azurea. In: Acta biologica Szegediensis, (61) 1. pp. 45-50. (2017)

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To establish the optimal cell disruption and protein extraction protocol for achieving the most efficient whole-cell protein extraction of Saccharomonospora azurea, four commonly used methods (X-Press, bead-vortexing, freezing-throwing and TCA/acetone/phenol extraction) were compared. Total protein content, as well as 1D and 2D SDS-PAGE protein patterns were assayed in the extracts to study the efficacy of these methods. Accordingly, of the four methods the X-Press proved the most effective for all initial weight (maximum 21.523 ± 0.23 mg/ml protein) followed by TCA/acetone/phenol method (maximum 13.682 ± 0.15 mg/ml protein), while the effectiveness of the two other methods were substantially inferior (maximum 3.188 ± 0.03 mg/ml protein). The analysis of protein gels proved that the X-Press method revealed a protein pattern characterised by the presence of the highest number of protein bands (on average of 52 and 385, on 1D and 2D gels, respectively). The TCA/acetone/phenol extraction provided similar effectiveness for only 100-300 mg initial bacteria mass, whereas bead-vortexing produced maximum 35 and 227 separated protein bands, on 1D and 2D gels, respectively. It can be stated that of the four methods the X-Press was the most effective one for all initial weight of bacteria, while the TCA/acetone/phenol method provides interpretable results for the 100-300 mg weight-range of bacteria.

Mű típusa: Cikk, tanulmány, mű
Egyéb cím: Protein extraction from Saccharomonospora azurea
Befoglaló folyóirat/kiadvány címe: Acta biologica Szegediensis
Dátum: 2017
Kötet: 61
Szám: 1
ISSN: 1588-4082
Oldalak: pp. 45-50
Befoglaló mű URL: http://acta.bibl.u-szeged.hu/49603/
Kulcsszavak: Biológia
Megjegyzések: Illusztrációval ; Bibliogr.: p. 49-50.
Feltöltés dátuma: 2017. dec. 06. 15:18
Utolsó módosítás: 2021. ápr. 12. 14:26
URI: http://acta.bibl.u-szeged.hu/id/eprint/49652
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