Metabolic engineering of Ralstonia eutropha for the production of polyhydroxyalkanoates from sucrose. Issue 3 (21st October 2014)

• Record Type: Journal Article
• Authors: 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
• Is Part Of: Biotechnology and bioengineering. Volume 112:Issue 3(2015:Mar.)
• Page Start: 638
• Page End: 643
• Publication Date: 2014-10-21
• Subjects: Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.25469
• Languages: English
• ISSNs: 0006-3592

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>ABSTRACT</title> <sec id="bit25469-sec-0001" sec-type="section"> <p>A sucrose utilization pathway was established in <italic>Ralstonia eutropha</italic> NCIMB11599 and <italic>R. eutropha</italic> 437‐540 by introducing the <italic>Mannheimia succiniciproducens</italic> MBEL55E <italic>sacC</italic> gene that encodes β‐fructofuranosidase. These engineered strains were examined for the production of poly(3‐hydroxybutyrate) [P(3HB)] and poly(3‐hydroxybutyrate‐<italic>co</italic>‐lactate) [P(3HB‐<italic>co</italic>‐LA)], respectively, from sucrose as a carbon source. It was found that β‐fructofuranosidase excreted into the culture medium could hydrolyze sucrose to glucose and fructose, which were efficiently used as carbon sources by recombinant <italic>R. eutropha</italic> strains. When <italic>R. eutropha</italic> NCIMB11599 expressing the <italic>sacC</italic> gene was cultured in nitrogen‐free chemically defined medium containing 20 g/L of sucrose, a high P(3HB) content of 73.2 wt% could be obtained. In addition, <italic>R. eutropha</italic> 437‐540 expressing the <italic>Pseudomonas</italic> sp. MBEL 6‐19 <italic>phaC1437</italic> gene and the <italic>Clostridium propionicum pct540</italic> gene accumulated P(3HB‐<italic>co</italic>‐21.5 mol% LA) to a polymer content of 19.5 wt% from sucrose by the expression of the <italic>sacC</italic> gene and the <italic>Escherichia coli ldhA</italic> gene. The molecular weights of P(3HB) and P(3HB‐<italic>co</italic>‐21.5 mol%LA) synthesized in <italic>R. eutropha</italic> using sucrose as a carbon source were 3.52 × 10<sup>5</sup> (M<sub>n</sub>) and 2.19 × 10<sup>4</sup> (M<sub>n</sub>), respectively. The engineered <italic>R. eutropha</italic> 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: 638–643. © 2014 Wiley Periodicals, Inc.</p> </sec> </abstract>