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首页> 外文期刊>Separation and Purification Technology >Partitioning of Geotrichum candidum Lipase from fermentative crude extract by aqueous two-phase system of polyethylene glycol and sodium citrate
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Partitioning of Geotrichum candidum Lipase from fermentative crude extract by aqueous two-phase system of polyethylene glycol and sodium citrate

机译：聚乙二醇和柠檬酸钠的水相两相法从发酵粗提物中分离白念珠菌脂肪酶

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The partitioning behaviour of the crude extracellular lipase of Geotrichum candidum (GCL) were studied in aqueous two-phase systems (ATPS) formed by mixtures of poly(ethylene oxide) (PEO) (1500 or 4000 or 6000 g mol(-1)) or triblock copolymer [(EO)(11)(PO)(16)(EO)(11)] (L35) (1900 g mol(-1)), where EO and PO are ethylene oxide and propylene oxide, respectively, + sodium citrate + water. The partition ratio for the proteins present in the crude broth (which is predominantly lipase) in the different ATPS were investigated as a function of the pH, PEO molar mass, phases hydrophobicity, tie-line length and temperature. The proteins were preferentially partitioned into the salt-rich phase in all of the systems studied with partition ratios ranging between 0.025 and 0.7. The thermodynamic parameters of transfers of the proteins between phases were obtained and shows that the transfer of proteins to the bottom phase is entropic driven, however negative values obtained for the enthalpy of the transfer of proteins the bottom to the top phase (Delta H-tr degrees = -12.12 to -7.10 kJ mol(-1) in the temperature range of 5.0-45.0 degrees C, respectively) constitutes evidence of the strong interactions between the proteins presents in the crude broth and the PEO chains in the top phase of the ATPS. The ATPS formed for a mixture of PEO 1500 g moll + sodium citrate + water at pH 7.0 was optimised using the response-surface methodology. The optimized conditions for the partition of proteins of crude broth were: 19.0% w/w + 12.0% w/w and 17.6% w/w + 14.0% w/w of PEO 1500 g moll and sodium citrate, respectively at pH 7.0. In these conditions were obtained the highest enzymatic concentrations in the bottom phase, with K-p < 0.15. (C) 2015 Elsevier B.V. All rights reserved.
机译：在由聚环氧乙烷（PEO）（1500或4000或6000 g mol（-1）的混合物形成的水性两相系统（ATPS）中研究了念珠菌（GCL）的粗胞外脂肪酶的分配行为或三嵌段共聚物[（EO）（11）（PO）（16）（EO）（11）]（L35）（1900 g mol（-1）），其中EO和PO分别为环氧乙烷和环氧丙烷，柠檬酸钠+水。研究了在不同ATPS中存在于粗肉汤（主要是脂肪酶）中的蛋白质的分配比率，该比率是pH，PEO摩尔质量，相疏水性，连接线长度和温度的函数。在所有研究的系统中，蛋白质都优先分配到富盐相中，分配比例在0.025到0.7之间。获得了蛋白质在各相之间转移的热力学参数，表明蛋白质向底部相的转移是由熵驱动的，但是从底部向顶部相的蛋白质转移的焓获得负值（ΔH-tr分别在5.0-45.0摄氏度的温度范围内= -12.12至-7.10 kJ mol（-1）构成了存在于粗肉汤中的蛋白质与PEO链在顶部相中的强相互作用的证据。 ATPS。使用响应表面方法优化了pH 7.0的PEO 1500克摩尔+柠檬酸钠+水的混合物形成的ATPS。粗肉汤蛋白的最佳分配条件为：pH 7.0时，PEO 1500 g摩尔和柠檬酸钠的含量分别为19.0％w / w + 12.0％w / w和17.6％w / w + 14.0％w / w。在这些条件下，在底部相中获得了最高的酶浓度，K-p <0.15。 （C）2015 Elsevier B.V.保留所有权利。

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