Cilt 15, Sayı 1, Sayfalar 8 - 16 2017-04-15

Aspergillus sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım
A Chromatographic Approach for Partial Purification of Polygalacturonase Produced by Aspergillus sojae

Ilknur Sen [1] , Marco A. Mata-Gomez [2] , Marco Rito-Palomares [3] , Canan Tari [4] , Melike Dinç [5]

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Bu çalışmanın amacı, A. sojae mutantından poligalakturonaz üretilmesi ve ham ekstraktın kromatografik yöntemlerle kısmi saflaştırılmasıdır. Peptitlerin konfirmasyonu için ilk basamak olarak, jel içinde sindirilmiş sodyum-dodesil-sülfat-poliakrilamid-jel-elektroforezi (SDS-PAGE) jellerinde matriks-yardımlı lazer desorpsiyon/iyonlaştırmalı-uçuş zamanlı-kütle spektrometresi (Maldi-TOF MS) analizi yapılmıştır. Poligalakturonaz üretimi için, katı-faz ve derin fermentasyonlarda üç farklı karbon kaynağı kullanılmıştır. Ham ekstrakt ilk olarak iyon değişim kromatografisi (IEXC) ile saflaştırılmıştır ve ardından bunu boyut eleme kromatografisi izlemiştir. Derin [acı portakal kabuğu, şeker pancarı melası ve (NH4)2SO4] ve katı-faz (buğday kepeği, şeker pancarı ve HCl) fermentasyonlarından elde edilen ham ekstraktlar yüksek seviyede poligalakturonaz enzim aktivitesi (sırasıyla 95.22 and 50.27 U/mL) göstermiştir. IEXC toplanmış fraksiyonunun (180, 200 ve 220 mM tuz fraksiyonları) boyut elemesi, en yüksek verimi (%36) ve saflaştırma katını (2.00) göstermiştir. SDS-PAGE’den elde edilen olası poligalakturonaz bantları jel içinde sindirilmiş ve peptit konfirmasyonu için Maldi-TOF-MS ile analiz edilmiştir. 

The aim of this study was to produce polygalacturonase from A. sojae mutant and partially purify the crude extract by chromatographic methods. As a preliminary step for the confirmation of its peptides, matrix-assisted laser-desorption-ionization-time-of-flight mass spectrometry (Maldi-TOF MS) analysis was performed on in-gel digested sodium-dodecyl-sulphate-polyacrylamide-gel-electrophoresis (SDS-PAGE) gels. Three different carbon sources were employed in submerged and solid-state fermentations for the production of polygalacturonase. Crude extract was first purified by ion-exchange chromatography (IEXC) and followed further by size exclusion chromatography. Crude extracts obtained from sub-merged [of bitter orange peel, sugar beet molasses and (NH4)2SO4] and solid-state [of wheat bran, sugar beet and HCl] fermentation exhibited high levels of polygalacturonase enzyme activity (95.22 and 50.27 U/mL, respectively). Size exclusion of IEXC pooled fraction (180, 200 and 220 mM salt fractions) revealed the highest yield (36%) and purification fold (2.00). The likely polygalacturonase bands from SDS-PAGE were in-gel digested and analyzed by Maldi-TOF MS in route for peptides confirmation.

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Konular
Dergi Bölümü Araştırma Makaleleri
Yazarlar

Yazar: Ilknur Sen
E-posta: ilknursen@iyte.edu.tr

Yazar: Marco A. Mata-Gomez
E-posta: ilknursen@iyte.edu.tr

Yazar: Marco Rito-Palomares
E-posta: mrito@itesm.mx

Yazar: Canan Tari
E-posta: canantari@iyte.edu.tr

Yazar: Melike Dinç
E-posta: melikedinc@iyte.edu.tr

Bibtex @araştırma makalesi { akademik-gida305767, journal = {Akademik Gıda}, issn = {1304-7582}, address = {Sidas Medya Ajans Tanıtım Danışmanlık Ltd. Şti.}, year = {2017}, volume = {15}, pages = {8 - 16}, doi = {10.24323/akademik-gida.305767}, title = {Aspergillus sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım}, language = {tr}, key = {cite}, author = {Dinç, Melike and Mata-Gomez, Marco A. and Rito-Palomares, Marco and Sen, Ilknur and Tari, Canan} } @araştırma makalesi { akademik-gida305767, journal = {Akademik Gıda}, issn = {1304-7582}, address = {Sidas Medya Ajans Tanıtım Danışmanlık Ltd. Şti.}, year = {2017}, volume = {15}, pages = {8 - 16}, doi = {10.24323/akademik-gida.305767}, title = {A Chromatographic Approach for Partial Purification of Polygalacturonase Produced by Aspergillus sojae}, language = {en}, key = {cite}, author = {Dinç, Melike and Mata-Gomez, Marco A. and Rito-Palomares, Marco and Sen, Ilknur and Tari, Canan} }
APA Sen, I , Mata-Gomez, M , Rito-Palomares, M , Tari, C , Dinç, M . (2017). Aspergillus sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım. Akademik Gıda, 15 (1), 8-16. DOI: 10.24323/akademik-gida.305767
MLA Sen, I , Mata-Gomez, M , Rito-Palomares, M , Tari, C , Dinç, M . "Aspergillus sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım". Akademik Gıda 15 (2017): 8-16 <http://dergipark.gov.tr/akademik-gida/issue/28716/305767>
Chicago Sen, I , Mata-Gomez, M , Rito-Palomares, M , Tari, C , Dinç, M . "Aspergillus sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım". Akademik Gıda 15 (2017): 8-16
RIS TY - JOUR T1 - Aspergillus sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım AU - Ilknur Sen , Marco A. Mata-Gomez , Marco Rito-Palomares , Canan Tari , Melike Dinç Y1 - 2017 PY - 2017 N1 - doi: 10.24323/akademik-gida.305767 DO - 10.24323/akademik-gida.305767 T2 - Akademik Gıda JF - Journal JO - JOR SP - 8 EP - 16 VL - 15 IS - 1 SN - 1304-7582-2148-015X M3 - doi: 10.24323/akademik-gida.305767 UR - http://dx.doi.org/10.24323/akademik-gida.305767 Y2 - 2017 ER -
EndNote %0 Akademik Gıda Aspergillus sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım %A Ilknur Sen , Marco A. Mata-Gomez , Marco Rito-Palomares , Canan Tari , Melike Dinç %T Aspergillus sojae Tarafından Üretilen Poligalakturonazın Kısmi Saflaştırılması için Kromatografik Bir Yaklaşım %D 2017 %J Akademik Gıda %P 1304-7582-2148-015X %V 15 %N 1 %R doi: 10.24323/akademik-gida.305767 %U 10.24323/akademik-gida.305767