Demir, Nazan; Nadaroglu, Hayrunnisa; Demir, Yasar; Isik, Ceyhun; Taskin, Esen; Adiguzel, Ahmet; Gulluce, Medine

An alkaline pectin lyase (Pnl) (EC 4.2.2.10) secreted by Brevibacillus borstelensis P35 (genBank number: FJ417406) was purifed using ammonium sulfate fractionation, anion exchange chromatography on DEaE-cellulose and gel fltration chromatography on Sephadex g-150. The pH and temperature optima of the enzyme were found to be 8.0 and 60 °C. The enzyme does not loose activity up to 60 °C if exposed for 1 h. The values of Kmand Vmax of the enzyme were 0.625 mg/ml and 126.32 s-1, respectively. The molecular weight was found to be 36 ± 01 kDa. The presence of 10 mM concentration of Ca2+, Cu2+, Mn2+, Mg2+, Zn2+, Hg2+, Fe2+ and EDTa, L-cystein, ascorbic acid signifcantly enhanced the Pnl of the purifed enzyme. In the course of the laboratory trials, it was demonstrated that Pnl from B. borstelensis (P35) could be successfully applied to the production and clarif-cation of fruit juice and oil extraction.

AtıfScopus36·WoS31·OpenAlex35

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Nadaroǧlu, Hayrunnisa; Taşkin, Esen; Adigüzel, Ahmet; Güllüce, Medine; Demir, Nazan

Extracellular pectin lyase (EC 4.2.2.10) was produced by Bacillus pumilus (P9) in solid state fermentation. Pectin lyase enzyme was purified 36.36 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 25 kDa by using Sephadex G-100 gel filtration chromatography. Purification of enzyme was controlled by SDS-PAGE. The optimum pH and temperature of enzyme was determined as pH 6.0 and 60oC, respectively. Pectin lyase was mostly stable at 40°C. Its' activity deceased in 50% for 1h at 60°C and 40% for 4 h at 50°C. Vmax and KM were calculated respectively as 0.298 mg/mL and 132.6 mol/L*min. The presence of 10 mM concentration of Ca2+, Cu2+, Mn2+, Mg2+, Zn2+, Hg2+, Fe2+ and EDTA, L-cystein, ascorbic acid and -mercaptoethanol significantly enhanced the pectin lyase of the purified enzyme. The purified pectin lyase enzyme was used for getting fruits juices. It was determined that yields of fruits juices significantly improved when it was compared with control.

AtıfScopus33

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