Dept. of Chemical Engineering, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil.
Biotechnol Prog. 2012 Nov-Dec;28(6):1575-80. doi: 10.1002/btpr.1624. Epub 2012 Sep 27.
Poly(3-hydroxybutyrate) (P(3HB)) and its copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(3HB-co-3HV), are biodegradable thermoplastic polymers. They are members of the polyhydroxyalkanoate (PHA) family, synthesized and accumulated as a carbon and energy reserve by a variety of microorganisms. The aim of this study was to evaluate the use of the proteases Corolase® L10, Alcalase® 2.4L, Corolase® 7089 and Protemax® FC and glycosidases Celumax® BC, Rohament® CL and Rohalase® Barley for the recovery of P(3HB) and P(3HB-co-3HV) synthesized by Cupriavidus necator. The enzyme Celumax® BC provided better lysis of the bacterial cell membrane and the results for the optimization of the operating conditions showed that this enzyme is most stable in acetate buffer at pH 4.0, bath at 60°C, hydrolysis time of 1 h and concentration of 0.02% (w/w). The optimization of the operating conditions showed that the enzyme Celumax® BC provided better lysis of the bacterial cell in acetate buffer at pH 4.0, bath at 60°C, hydrolysis time of 1 h and concentration of 0.02% (w/w). These conditions resulted in lysis of the membrane of the bacteria with a recovery of 93.2% P(3HB-co-3HV) with 94% purity. The results showed that the use of enzymes for the polymer extraction is an efficient process that assists in the cell disruption of Cupriavidus necator.
聚（3-羟基丁酸酯）（P（3HB））及其共聚物聚（3-羟基丁酸酯-共-3-羟基戊酸酯）（P（3HB-co-3HV）是可生物降解的热塑性聚合物。它们是聚羟基烷酸酯（PHA）家族的成员，由各种微生物合成并积累为碳和能量储备。本研究的目的是评估蛋白酶 Corolase® L10、Alcalase® 2.4L、Corolase® 7089 和 Protemax® FC 以及糖苷酶 Celumax® BC、Rohament® CL 和 Rohalase® Barley 用于回收 Cupriavidus necator 合成的 P（3HB）和 P（3HB-co-3HV）的使用。酶 Celumax® BC 提供了更好的细菌细胞膜裂解，优化操作条件的结果表明，该酶在 pH4.0 的醋酸盐缓冲液中最稳定，浴温 60°C，水解时间 1 小时，浓度为 0.02%（w/w）。优化操作条件的结果表明，酶 Celumax® BC 在 pH4.0 的醋酸盐缓冲液中提供了更好的细菌细胞膜裂解，浴温 60°C，水解时间 1 小时，浓度为 0.02%（w/w）。这些条件导致膜的裂解，93.2%的 P（3HB-co-3HV）的回收率为 94%纯度。结果表明，使用酶进行聚合物提取是一种有效的工艺，有助于 Cupriavidus necator 的细胞破裂。
