通过改变介质的离子强度并用醇沉淀来纯化菊粉酶。
Purification of inulinases by changing the ionic strength of the medium and precipitation with alcohols.
作者信息
Golunski Simone, Silva Marceli F, Marques Camila T, Rosseto Vanusa, Kaizer Rosilene R, Mossi Altemir J, Rigo Diane, Dallago Rogério M, DI Luccio Marco, Treichel Helen
机构信息
Universidade Regional Integrada do Alto Uruguai e das Missões/URI, Departamento de Engenharia de Alimentos, Campus de Erechim, Av. 7 de Setembro, 1621, 99700-000 Erechim, RS, Brazil.
Programa de Pós-Graduação em Ciência e Tecnologia Ambiental, Universidade Federal da Fronteira Sul, Campus de Erechim, Rodovia ERS 135, Km 72, 99700-970 Erechim, RS, Brazil.
出版信息
An Acad Bras Cienc. 2017 Jan-Mar;89(1):57-63. doi: 10.1590/0001-3765201720160367.
The present study evaluated the purification of inulinase by changing the ionic strength of the medium by addition of NaCl and CaCl2 followed by precipitation with n-propyl alcohol or iso-propyl alcohol. The effects of the concentration of alcohols and the rate of addition of alcohols in the crude extract on the purification yield and purification factor were evaluated. Precipitation caused an activation of enzyme and allowed purification factors up to 2.4-fold for both alcohols. The purification factor was affected positively by the modification of the ionic strength of the medium to 0.5 mol.L-1 NaCl before precipitation with the alcohol (n-propyl or iso-propyl). A purification factor of 4.8-fold and an enzyme yield of 78.1 % could be achieved by the addition of 0.5 mol.L-1 of NaCl to the crude extract, followed by the precipitation with 50 % (v/v) of n-propyl alcohol, added at a flow rate of 19.9 mL/min.
本研究通过添加氯化钠和氯化钙改变培养基的离子强度，随后用正丙醇或异丙醇沉淀，评估菊粉酶的纯化效果。评估了粗提物中醇的浓度和醇的添加速率对纯化产率和纯化因子的影响。沉淀导致酶的激活，两种醇的纯化因子可达2.4倍。在用醇（正丙醇或异丙醇）沉淀之前，将培养基的离子强度调节至0.5 mol.L-1氯化钠，对纯化因子有积极影响。通过向粗提物中添加0.5 mol.L-1氯化钠，随后用50%（v/v）正丙醇以19.9 mL/min的流速添加进行沉淀，可实现4.8倍的纯化因子和78.1%的酶产率。
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