Purification process and characterization of Naringinase from Penicillium sp.1523

Purification process of naringinase from Penicillium sp.1523 was explored using ultrafiltration, ammonium sulfate precipitation and Sephadex G-200 gel filtration chromatography methods, and the purification effects were identified by SDS-PAGE experiment at last.The purification protocol resulted in 11.99-fold purified naringinase with a final yield of 38.10%, and the specific activity reached 4478.76 U /mg.After successive purification two clear protein bands were obtained by SDS-PAGE, which indicated naringinase consisted of two subnits with molecular masses of 89 ku and 72 ku respectively.The temperature and pH optima for this enzyme were 50℃ and 4.0, and naringinase stability study showed a lower thermal stability. Ca2 +, Mg2 +, Mn2 +, Zn2 +, EDTA and Cu2 +could all lead to the increase of naringinase activity while Fe2 +and SDS showed inhibition of naringinase even at lower concentration.This study can supply an experimental basis for further scale up of naringinase catalysis and molecular modification.