Production of pectin lyase from Geobacillus pallidus p26, purifi cation, characterization and fruit juice application
Safinur Yildirim Çelik, Nazan Demir a, Yaşar Demir b, Ahmet Adiguzel c, Medine Gulluce d
College of Education, Bayburt University, 69000, Bayburt, Turkey
a College of Art and Science, Muğla University, 48000, Muğla, Turkey
b College of Art and Science, Muğla University, 48000, Muğla, Turkey
c Atatürk University, Faculty of Science, Molecular Biology and Genetics Department, 25240, Erzurum, Turkey
d Atatürk University, Faculty of Science, Biology Department, 25240, Erzurum, Turkey
Abstract: A bacterial strain was isolated from Pasinler hot spring, Erzurum, Turkey. The purified thermophilic isolate was identified as Geobacillus pallidus P26 and used to produce extracellular pectin lyase (EC 4.2.2.10). Pectin lyase enzyme was purified 34 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 56 kDa by using Sephadex G-100 gel filtration chromatography. Purification of enzyme was verified by SDS-PAGE. The pH- and temperature optima of enzyme were determined (pH 9.0 and 60oC, respectively). Pectin lyase was mostly stable at 50oC for 24 hours. Its’ activity decreased to 50% for 24 h at 60oC. KM and Vmax were calculated as 24.8 mg/mL and 2.28 μmol/L*min, respectively. Purified pectin lyase was inhibited by Fe3+, Zn2+, Cu2+, Ca2+, Co2+ and Hg2+ but not by Mg2+. The purified pectin lyase enzyme was used for getting fruits juices. It was found that yields of fruits juices increased when they were compared with control.
Keywords: Pectin lyase, purification, characterization, Geobacillus pallidus P26
Full paper in Portable Document Format: acs_0187.pdf
Acta Chimica Slovaca, Vol. 7, No. 1, 2014, pp. 57—63, DOI: 10.2478/acs-2014-0011