Pre-separation and pre-concentration of bacteria is an important step especially when they are uncultured and bacterial concentration in the matrix is low. This study describes a preparative method based on isoelectric focusing of colored microorganisms in a cellulose-based separation medium from a high conductivity matrix. The isoelectric points found for the examined cells were 1.8 for Micrococcus luteus, 3.5 for Dietzia sp., and 4.7 for Rhodotorula mucilaginosa using capillary isoelectric focusing. The final positions of the zones of colored microbial cells in the cellulose-bed are indicated by colored pI markers. Segments of the separation medium with cells were harvested by a spatula, simply purified using centrifugation and analyzed by capillary isoelectric focusing and matrix-assisted laser desorption/ionization time of flight mass spectrometry. The determined recovery ranged from 78% to 93%. The viability of the harvested cells was verified by their cultivation. Copyright © 2017 Elsevier B.V. All rights reserved.

Horka Marie Institute of Analytical Chemistry of the CAS v v i Veveri 97 602 00 Brno Czech Republic

000      

00000naa a2200000 a 4500

001      

bmc22005051

003      

CZ-PrNML

005      

20220404161111.0

007      

ta

008      

220118s2017 ne f 000 0|eng||

009      

AR

024    7_

$a 10.1016/j.aca.2017.08.046 $2 doi

035    __

$a (PubMed)29029742

040    __

$a ABA008 $b cze $d ABA008 $e AACR2

041    0_

$a eng

044    __

$a ne

100    1_

$a Horká, Marie $7 xx0076574 $u Horka, Marie. Institute of Analytical Chemistry of the CAS, v. v. i., Veveri 97, 602 00 Brno, Czech Republic. Electronic address: horka@iach.cz.

245    10

$a Preparative isoelectric focusing of microorganisms in cellulose-based separation medium and subsequent analysis by CIEF and MALDI-TOF MS / $c M Horka, K Slais, J Salplachta, F Ruzicka

520    9_

$a Pre-separation and pre-concentration of bacteria is an important step especially when they are uncultured and bacterial concentration in the matrix is low. This study describes a preparative method based on isoelectric focusing of colored microorganisms in a cellulose-based separation medium from a high conductivity matrix. The isoelectric points found for the examined cells were 1.8 for Micrococcus luteus, 3.5 for Dietzia sp., and 4.7 for Rhodotorula mucilaginosa using capillary isoelectric focusing. The final positions of the zones of colored microbial cells in the cellulose-bed are indicated by colored pI markers. Segments of the separation medium with cells were harvested by a spatula, simply purified using centrifugation and analyzed by capillary isoelectric focusing and matrix-assisted laser desorption/ionization time of flight mass spectrometry. The determined recovery ranged from 78% to 93%. The viability of the harvested cells was verified by their cultivation.<ovid:br/><ovid:br/> Copyright &#xa9; 2017 Elsevier B.V. All rights reserved.

650    12

$a Actinobacteria $x izolace a purifikace $7 D039903

650    12

$a celulosa $x chemie $7 D002482

650    12

$a isoelektrická fokusace $7 D007525

650    12

$a Micrococcus $x izolace a purifikace $7 D008837

650    12

$a Rhodotorula $x izolace a purifikace $7 D012248

650    12

$a spektrometrie hmotnostní - ionizace laserem za účasti matrice $7 D019032

700    1_

$a Šlais, Karel $7 xx0071057

700    1_

$a Šalplachta, Jiří

700    1_

$a Růžička, Filip, $d 1972- $7 xx0071619

773    0_

$t Analytica Chimica Acta $g č. 990 (2017), s. 185-193 $p Anal Chim Acta $x 1873-4324 $w MED00000332

773    0_

$p Anal Chim Acta $g 990:185-193, 2017 Oct 16

910    __

$a ABA008 $y p $z 0 $b sig

990    __

$a 20220118155405 $b ABA008

991    __

$a 20220404161109 $b ABA008

999    __

$a ok $b bmc $g 1747510 $s 1156200

BAS    __

$a 3

BMC    __

$a 2017 $c 990 $d 185-193 $x MED00000332 $i 1873-4324 $m Analytica chimica acta

GRA    __

$a NV16-29916A $p MZ0

LZP    __

$a 2021-granty