Separation of the protease enzymes of Bacillus licheniformis from the fermentation medium by crossflow ultrafiltration

Abstract

The separation from fermentation medium of extracellular serine alkaline protease (SAP) enzyme produced by Bacillus licheniformis was investigated using a crossflow ultrafiltration system. SAP was separated from the high molecular weight neutral protease (NP) and amylase (AMY) enzymes and from the low molecular weight organic acids and amino acids in a crossflow ultrafiltration system with 30000 Da and 10000 Da MWCO polysulfone membranes, respectively. The effects of transmembrane pressure (TMP), recirculation velocity (v), and initial enzyme concentration (CE) on the permeate flux, on the activities of SAP, NP and AMY enzymes, and on the recovery of SAP were investigated. High permeate flux was obtained at high recirculation velocity and TMP, but at low initial enzyme concentration. SAP enzyme recovery and activity measured in the system also showed alterations with hydrodynamic conditions. The best operation conditions for the separation of SAP from NP and AMY were TMP = 20 kPa, v = 0.50 ms(-1) and C-E = 0.2 gdm(-3). The separation of SAP from the organic and amino acids was best performed at TMP = 100 kPa, v = 0.33 ms(-1) and C-E = 0.40 gdm(-3). (C) 2000 Society of Chemical Industry.