Construction of genetically engineered Candida tropicalis for conversion of l-arabinose to l-ribulose

•We constructed an l-ribulose-producing Candida tropicalis with co-expression of transporter and l-arabinose isomerase genes.•Increased l-arabinose uptake was achieved by expressing transporter, STP2 from A. thaliana.•Expression of l-arabinose isomerase from L. sakei 23K showed the highest conversio...

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Veröffentlicht in:Journal of biotechnology 2018-05, Vol.274, p.9-14
Hauptverfasser: Yeo, In-Seok, Shim, Woo-Yong, Kim, Jung Hoe
Format: Artikel
Sprache:eng
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Zusammenfassung:•We constructed an l-ribulose-producing Candida tropicalis with co-expression of transporter and l-arabinose isomerase genes.•Increased l-arabinose uptake was achieved by expressing transporter, STP2 from A. thaliana.•Expression of l-arabinose isomerase from L. sakei 23K showed the highest conversion yield.•A conversion yield of 30% was achieved by the engineered strain from a glucose/l-arabinose mixture. For the biological production of l-ribulose, conversion by enzymes or resting cells has been investigated. However, expensive or concentrated substrates, an additional purification step to remove borate and the requirement for cell cultivation and harvest steps before utilization of resting cells make the production process complex and unfavorable. Microbial fermentation may help overcome these limitations. In this study, we constructed a genetically engineered Candida tropicalis strain to produce l-ribulose by fermentation with a glucose/l-arabinose mixture. For the uptake of l-arabinose as a substrate and conversion of l-arabinose to l-ribulose, two heterologous genes coding for l-arabinose transporter and l-arabinose isomerase, were constitutively expressed in C. tropicalis under the GAPDH promoter. The Arabidopsis thaliana-originated l-arabinose transporter gene (STP2)-expressing strain exhibited a high l-arabinose uptake rate of 0.103 g/g cell/h and the expression of l-arabinose isomerase from Lactobacillus sakei 23 K showed 30% of conversion (9 g/L) from 30 g/L of l-arabinose. This genetically engineered strain can be used for l-ribulose production by fermentation using mixed sugars of glucose and l-arabinose.
ISSN:0168-1656
1873-4863
DOI:10.1016/j.jbiotec.2018.01.019