Integration of the production and the purification processes of cutinase secreted by a recombinant Saccharomyces cerevisiae SU50 strain
Integration of the production and the purification processes of cutinase secreted by a recombinant Saccharomyces cerevisiae SU50 strain
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DOI：
10.1016/j.jbiotec.2003.10.032
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发表时间：
2004-04-08
影响因子：
4.1
通讯作者：
Fonseca, LP
中科院分区：
文献类型：
--
作者：
Calado, CRC;Ferreira, BS;Fonseca, LP
通过膨胀床吸附（EBA），可以同时直接从粗原料中回收和纯化异源角质酶。然而，据观察，在高度浓缩且因此具有经济优势的EBA纯化过程中，培养步骤高度影响后续纯化步骤。因此，酵母培养和EBA角质酶纯化不能被视为独立的实体，了解它们之间的相互作用对于开发高成本效益的整体角质酶生产工艺至关重要。从研究的培养策略（一批、一次连续和两次补料分批培养）来看，导致更经济的角质酶整体生产工艺的策略是补料分批模式，并添加半乳糖。最后一种培养策略表现出最高的培养角质酶活性和生物反应器生产率，与以葡萄糖和半乳糖为补料的分批补料培养相比，角质酶活性提高了 3.8 倍，生产率提高了 3.0 倍，可以补偿高出 40% 的培养基成本。此外，与以葡萄糖和半乳糖补料的分批补料培养相比，获得了3.8倍的有效角质酶动态吸附能力和3.8倍的有效纯化生产率。以半乳糖为食的培养策略，其有效纯化效率提高了 5.6 倍，还可以补偿连续培养肉汤获得的 32% 有效吸附能力。此外，与连续培养相比，角质酶活性提高了 205 倍，生物反应器生产率提高了 24 倍，培养基成本降低了 6%。 (C) 2004 Elsevier B.V. 保留所有权利。
By expanded bed adsorption (EBA) it was possible to simultaneously recover and purify the heterologous cutinase directly from the crude feedstock. However, it was observed that in a highly condensed and consequently economically advantageous purification process as EBA, the cultivation step highly influences the following purification step. Thus, the yeast cultivation and cutinase purification by EBA cannot be considered as independent entities, and the understanding of the interactions between them are crucial for the development of a highly cost effective overall cutinase production process.From the cultivation strategies studied, one batch, one continuous and two fed-batch cultivations, the strategy that resulted in a more economical cutinase overall production process was a fed-batch mode with a feeding in galactose. This last cultivation strategy, exhibited the highest culture cutinase activity and bioreactor productivity, being obtained 3.8-fold higher cutinase activity and 3.0-fold higher productivity that could compensate the 40% higher cultivation medium costs when compared with a fed-batch culture with a feeding on glucose and galactose. Moreover, a 3.8-fold higher effective cutinase dynamic adsorption capacity and 3.8-fold higher effective purification productivity were obtained in relation to the fed-batch culture with the feeding on glucose and galactose. The cultivation strategy with a feeding on galactose, that presented 5.6-fold higher effective purification productivity, could also compensate the 32% effective adsorption capacity obtained with a continuous cultivation broth. Furthermore, a 205-fold higher cutinase activity, 24-fold higher bioreactor productivity and 6% of the cultivation medium costs were obtained in relation to the continuous culture. (C) 2004 Elsevier B.V. All rights reserved.