Partitioning and purification of the cellulolytic complex produced by Aspergillus japonicus URM5620 using PEG-citrate in an aqueous two-phase system
Partitioning and purification of the cellulolytic complex produced by Aspergillus japonicus URM5620 using PEG-citrate in an aqueous two-phase system
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DOI：
10.1016/j.fluid.2012.08.008
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发表时间：
2012-12-15
影响因子：
2.6
通讯作者：
Porto, Ana L. F.
中科院分区：
文献类型：
--
作者：
Herculano, Polyanna N.;Porto, Tatiana S.;Porto, Ana L. F.
研究了日本曲霉URM5620产纤维素水解复合物中纤维素酶在水两相体系（ATPS）中的分配和纯化。采用因子设计模型（2(4)）评价聚乙二醇（PEG）的摩尔质量（1000- 8000g /mol）、PEG浓度（20.0-24.0%）的影响。w/w)、柠檬酸钠（15-20%,w/w）浓度和pH（6.0-8.0）对β -葡萄糖苷酶（PG）组成的纤维素水解复合物的差异分配和纯化的影响。内切葡聚糖酶（CMCase）和总纤维素酶（FPase）。β G优先分配到盐相；而当PEG的摩尔质量和浓度同时增加时，分配系数增大（K=3.0）。在pH 8.0条件下，20% (w/w) PEG 8000 （g/mol）和10% （w/w）柠檬酸钠的处理效果最好；分配系数约为2.9，活性得率为77%，纯化系数为1.9。FPase和CMCase优先划分到顶部阶段。当顶相为PEG 1000 （g/mol）时，K值最高，分别为2.9和3.0。在pH 8.0条件下，PEG 1000 （g/mol）用量为24% （w/w）、柠檬酸钠用量为15% （w/w）时，FPase的最佳活性率为2.67%。20% (w/w) PEG 8000 （g/mol）时，纯化系数最高，为64.8。当pH 7.0时，PEG 3350 （g/mol）用量为22% (w/w)，柠檬酸钠用量为17.5% (w/w)， CMCase的活性产率为1.64%，纯化因子为45.9。PEG/柠檬酸钠体系导致纤维素酶的分离特性，其中β G分裂到盐相，而FPase和CMCase分裂到顶相。这个过程提供了一个有效的和有吸引力的增加净化系数。(C) 2012 Elsevier B.V.版权所有
The partitioning and purification of cellulases from the cellulolytic complexes produced by Aspergillus japonicas URM5620 were investigated in aqueous two-phase systems (ATPS). A factorial design model (2(4)) was used to evaluate the influence of polyethylene glycol (PEG) molar mass (1000-8000 g/mol), PEG concentration (20.0-24.0%. w/w), sodium citrate (15-20%, w/w) concentration and pH (6.0-8.0) on the differential partitioning and purification of the cellulolytic complex composed of beta-glucosidase (PG). endoglucanase (CMCase) and total cellulase (FPase). The beta G preferentially partitioned to the salt phase; however, when the molar mass and concentration of PEG were simultaneously increased, the partition coefficient increased (K=3.0). For beta G. 20% (w/w) PEG 8000 (g/mol) and 10% (w/w) sodium citrate at pH 8.0 produced the best results; the partition coefficient was approximately 2.9, the activity yield was 77%, and the purification factor was 1.9. The FPase and CMCase preferentially partitioned to the top phase. The highest K values (2.9 and 3.0) were obtained when the top phase consisted of PEG 1000 (g/mol). The best activity yield for FPase (2.67%) was obtained with 24% (w/w) PEG 1000 (g/mol) and 15% (w/w) sodium citrate at pH 8.0. However, the highest purification factor (64.8) was observed with 20% (w/w) PEG 8000 (g/mol). For CMCase, the highest activity yield (1.64%) and purification factor (45.9) were obtained with 22% (w/w) PEG 3350 (g/mol) and 17.5% (w/w) sodium citrate at pH 7.0. The PEG/sodium citrate system resulted in separations characteristic of the cellulases of cellulolytic complexes, where beta G partitions to the salt phase and FPase and CMCase partition to the top phase. This process provides an efficient and attractive increase in the purification factor. (C) 2012 Elsevier B.V. All rights reserved.