Construction of a stable plasmid vector for industrial production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by a recombinant Cupriavidus聽necator H16 strain

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• 作者：Shunsuke Sato ; Tetsuya Fujiki ; Keiji Matsumoto
• 关键词：Polyhydroxyalkanoate ; Ralstonia eutropha ; Cupriavidus metallidurans ; Stable plasmid vector ; Plasmid partition
• 刊名：Journal of Bioscience and Bioengineering
• 出版年：December, 2013
• 年：2013
• 卷：116
• 期：6
• 页码：677-681
• 全文大小：714 K

文摘

A new stable plasmid vector (pCUP3) was developed for high and stable production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH) using Cupriavidus necator H16 as the host strain. In pCUP3, it was found that the plasmid partition and replication region of the megaplasmid pMOL28 in the Cupriavidus metallidurans CH34 strain plays an important role in plasmid stability in C.聽necator H16. Moreover, the partition locus (comprising parA28 and parB28 and the parS28 region) is essential for plasmid maintenance under high-PHBH-accumulation. PHBH productivity by the C.聽necator H16/ds strain (phaC1 deactivated mutant strain) harboring a phaC_Ac NSDG within pCUP3 was identical to the productivity of poly(3-hydroxybutyrate) by the C.聽necator H16 strain when palm kernel oil was used as the sole carbon source without any antibiotics. This new vector is important for industrial mass production of polyhydroxyalkanoates using the C.聽necator H16 strain as the host, dispensing the necessity of the application of selective pressure such as antibiotics.