Acuña-Argüelles M E, Gutiérrez-Rojas M, Viniegra-González G, Favela-Torres E
Departamento de Biotecnologia, Universidad Autonoma, D.F., México.
Appl Microbiol Biotechnol. 1995 Oct;43(5):808-14. doi: 10.1007/BF02431912.
Three extracellular pectinases were produced by Aspergillus niger CH4 by submerged and solid-state fermentation, and their physicochemical and kinetic properties were studied. The highest productivities of endo- and exo-pectinase and pectin lyase were obtained with solid-state fermentation. The kinetic and physicochemical properties of these enzymes were influenced by the type of culture method used. All activities were very different in terms of pH and temperature optima, stability at different pH and temperature values and affinity for the substrate (Km values). In solid-state fermentation, all pectinase activities were more stable at extreme pH and temperature values but the Km values of endo-pectinase and pectin lyase were higher with respect to those activities obtained by the submerged-culture technique. The pectin lyase activity obtained by the submerged-culture technique showed substrate inhibition but the enzyme obtained by solid-state fermentation did not. Electrophoresis, using sodium dodecyl sulphate/polyacrylamide gel with enzymatic extracts obtained for both culture methods, showed the same number of protein bands but some differences were found in their electrophoretic position. The results obtained in this work suggest that the culture method (submerged or solid-state) may be responsible for inducing changes in some of the pectinolytic enzymes produced by A. niger.
黑曲霉CH4通过深层发酵和固态发酵产生了三种胞外果胶酶,并对其理化性质和动力学性质进行了研究。固态发酵获得了最高的内切和外切果胶酶及果胶裂解酶产量。这些酶的动力学和理化性质受所用培养方法类型的影响。所有酶活性在最适pH和温度、不同pH和温度值下的稳定性以及对底物的亲和力(Km值)方面都有很大差异。在固态发酵中,所有果胶酶活性在极端pH和温度值下更稳定,但内切果胶酶和果胶裂解酶的Km值相对于深层培养技术获得的那些活性更高。深层培养技术获得的果胶裂解酶活性表现出底物抑制,但固态发酵获得的酶没有。使用两种培养方法获得的酶提取物,通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳显示出相同数量的蛋白条带,但在其电泳位置上发现了一些差异。这项工作获得的结果表明,培养方法(深层或固态)可能导致黑曲霉产生的一些果胶分解酶发生变化。