BioChem - DOS

Metabolic engineering of Ralstonia eutropha for the production of polyhydroxyalkanoates from sucrose

Biotechnology and bioengineering

Park, Si Jae; Jang, Young‐ Ah; Noh, Won; Oh, Young Hoon; Lee, Hyuk; David, Yokimiko; Baylon, Mary Grace; Shin, Jihoon; Yang, Jung Eun; Choi, So Young; Lee, Seung Hwan; Lee, Sang Yup

<P><B>ABSTRACT</B></P><P>A sucrose utilization pathway was established in <I>Ralstonia eutropha</I> NCIMB11599 and <I>R. eutropha</I> 437&#8208;540 by introducing the <I>Mannheimia succiniciproducens</I> MBEL55E <I>sacC</I> gene that encodes &beta;&#8208;fructofuranosidase. These engineered strains were examined for the production of poly(3&#8208;hydroxybutyrate) [P(3HB)] and poly(3&#8208;hydroxybutyrate&#8208;<I>co</I>&#8208;lactate) [P(3HB&#8208;<I>co</I>&#8208;LA)], respectively, from sucrose as a carbon source. It was found that &beta;&#8208;fructofuranosidase excreted into the culture medium could hydrolyze sucrose to glucose and fructose, which were efficiently used as carbon sources by recombinant <I>R. eutropha</I> strains. When <I>R. eutropha</I> NCIMB11599 expressing the <I>sacC</I> gene was cultured in nitrogen&#8208;free chemically defined medium containing 20&thinsp;g/L of sucrose, a high P(3HB) content of 73.2&thinsp;wt% could be obtained. In addition, <I>R. eutropha</I> 437&#8208;540 expressing the <I>Pseudomonas</I> sp. MBEL 6&#8208;19 <I>phaC1437</I> gene and the <I>Clostridium propionicum pct540</I> gene accumulated P(3HB&#8208;<I>co</I>&#8208;21.5&thinsp;mol% LA) to a polymer content of 19.5&thinsp;wt% from sucrose by the expression of the <I>sacC</I> gene and the <I>Escherichia coli ldhA</I> gene. The molecular weights of P(3HB) and P(3HB&#8208;<I>co</I>&#8208;21.5&thinsp;mol%LA) synthesized in <I>R. eutropha</I> using sucrose as a carbon source were 3.52&thinsp;&times;&thinsp;10<SUP>5</SUP> (M<SUB>n</SUB>) and 2.19&thinsp;&times;&thinsp;10<SUP>4</SUP> (M<SUB>n</SUB>), respectively. The engineered <I>R. eutropha</I> strains reported here will be useful for the production of polyhydroxyalkanoates (PHAs) from sucrose, one of the most abundant and relatively inexpensive carbon sources. Biotechnol. Bioeng. 2015;112: 638&ndash;643. &copy; 2014 Wiley Periodicals, Inc.</P>

2015

0006-3592

1097-0290

112

3

pp.638-643

10.1002/bit.25469

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART71356147

β&#x2010; fructofuranosidase; poly(3&#x2010; hydroxybutyrate); poly(3&#x2010; hydroxybutyrate&#x2010; co&#x2010; lactate); Ralstonia eutropha; sucrose;