公开(公告)号：US5229279A

公开(公告)日：1993-07-20

申请号：US566535

申请日：1990-08-13

申请人： Oliver P. Peoples ,  Anthony J. Sinskey

发明人： Oliver P. Peoples ,  Anthony J. Sinskey

IPC分类号： C12N9/00 ,  C12N9/04 ,  C12N9/10 ,  C12N15/52 ,  C12P7/62

CPC分类号： C12N9/0006 ,  C12N15/52 ,  C12N9/00 ,  C12N9/1029 ,  C12P7/625

摘要： The present invention invention is a method for controlling biopolymer synthesis by determining the genetics and enzymology of polyhydroxybutyrate (PHB) biosynthesis at the molecular level. The purified enzymes and genes provide the means for developing new PHB-like biopolymers having polyester backbones. Specific aims are to 1) control the chain length of the polymers produced in fermentation processes through genetic manipulation, 2) incorporate different monomers into the polymers to produce copolymers with different physical properties, and 3) examine the physical/ rheological properties of these new biopolymers in order to develop further design criteria at the molecular level.The method for engineering biopolymer synthesis includes: isolation and characterization of the genes for the enzymes in the synthetic pathway (beta-ketothiolase, acetoacetyl-CoA reductase and PHB synthetase); cloning of the genes in a vector(s); placement of the vector(s) under the control of regulated promoters; expression of the genes; determination of the function and use of other factors such as substrate specificity in polymer production and composition; and isolation and physical and chemical analysis of the resulting polymers.

摘要翻译： 本发明是通过在分子水平上测定聚羟基丁酸（PHB）生物合成的遗传学和酶学方法来控制生物聚合物合成的方法。 纯化的酶和基因为开发具有聚酯主链的新型PHB样生物聚合物提供了手段。 具体目的是1）通过遗传操作控制在发酵过程中产生的聚合物的链长; 2）将不同单体引入聚合物中以产生具有不同物理性质的共聚物，以及3）检查这些新生物聚合物的物理/流变性质 以便在分子水平上制定进一步的设计标准。 工程生物聚合物合成的方法包括：合成途径中酶的分离和表征（β-酮硫解酶，乙酰乙酰辅酶A还原酶和PHB合成酶）; 在载体中克隆基因; 将载体置于受调节启动子的控制下; 基因表达; 确定聚合物生产和组成中其他因素如底物特异性的功能和用途; 以及所得聚合物的分离和物理和化学分析。