Preparation and Enzymatic Properties of Aspergillus niger Endoinulinase Immobilized onto Various Polysaccharide Supports.

Abstract

An extracellular endoinulinase P-III from Aspergillus niger 12 has been immobilized covalently onto bromoacetyl-cellulose (BAC), cellulose-carbonate (CC) and CNBr-activated Sepharose 4B (CAS) with the activity yields of 1 1.9, 24.1 and 19.3 % , respectively, under optimal reaction conditions for the immobilization. All three immobilized enzymes increased the resistance to inactivation by p-chloromercuribenzoate or Fe3'. Mn2. stimulated the activities of the immobilized endoinulinases onto CC (160 %) and CAS (168 %) more remarkably than those of free and BACimmobilized enzymes (124 %). The marked rise from 45 to 60'C in the optimal temperature for inulin hydrolysis was observed in the CAS-immobilized enzyme. The pH stability decreased on the acidic side and increased slightly on the alkaline side in the BAC-inunobilized enzyme, and shifted to the acidic side in the CC-immobilized enzyme. The K~ value decreased in the BAC-immobilized enzyme and increased in the CAS-immobilized enzyme.

Chemical and Process Engineering

“…Experimental literature data (Karimi et al, 2014;Megazyme, 2020;Nguyen et al, 2011;Zaita et al, 2000) were analysed for endo-inulinase from A. niger from various sources, including the commercial A. niger endo-inulinase from company Megazyme (Megazyme, 2020;Nguyen et al, 2011). Table 1 shows the conditions for measuring insulin activity, such as pH, buffer type and time measurement.…”

“…Measurements of inulinase activity were determined using dinitrosalicylic acid (Karimi et al, 2014;Zaita et al, 2000) at 575 nm and the Nelson-Somogyi method at 620 nm, (Megazyme, 2020;Nguyen et al, 2011). The initial inulin concentration measured was in the range of 0.2% to 6%.…”

“…The initial inulin concentration measured was in the range of 0.2% to 6%. Karimi et al (2014) and Zaita et al (2000) studied endo-inulinase activity during the hydrolysis of inulin buffered with various buffers and at various pH values for 30 minutes. Megazyme International Ireland (Ireland) (Megazyme, 2020) A. niger endo-inulinase activity was determined at a pH of 4.5 and a reaction time of 10 min.…”

“…There have been many publications in the literature on A. niger endo-inulinase (Karimi et al, 2014;Megazyme, 2020;Nguyen et al 2011;Zaita et al, 2000) used for immobilization process. However, processes involving endo-inulinase A. niger for industrial purposes cannot be designed and optimized without knowing the kinetic parameters of the process and thus the effect of temperature on activity endoinulinase (Ricca et al, 2009).…”

“…The work was aimed at modelling and determining parameters based on the available literature data. The method of determining the optimum temperatures and activation energies based on experimental data on the effect of temperature on the activity of endoinulinase A. niger has been presented by Karimi et al (2014), Megazyme, 2020, Nguyen et al (2011 or Zaita et al (2000).…”