l-Arabinose pathway engineering for arabitol-free xylitol production in Candida tropicalis

Yoon, Byoung Hoon; Jeon, Woo Young; Shim, Woo Yong; Kim, Jung Hoe

doi:10.1007/s10529-010-0487-2

Cited by 19 publications

(9 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the one-pot biotransformation has some unique advantages, constructing a “perfect” strain by metabolic engineering for monoculture is always attractive. In a previous report, the bacteria arabinose metabolic pathway was introduced into XYL2 -disrupted C. tropicalis [ 32 ]. The resulting recombinant strain converted xylose into xylitol, without arabitol formation.…”

Section: Discussionmentioning

confidence: 99%

Stepwise metabolic engineering of Candida tropicalis for efficient xylitol production from xylose mother liquor

et al. 2021

View full text Add to dashboard Cite

Background Commercial xylose purification produces xylose mother liquor (XML) as a major byproduct, which has become an inexpensive and abundant carbon source. A portion of this XML has been used to produce low-value-added products such as caramel but the remainder often ends up as an organic pollutant. This has become an issue of industrial concern. In this study, a uracil-deficient Candida tropicalis strain was engineered to efficiently convert XML to the commercially useful product xylitol. Results The xylitol dehydrogenase gene was deleted to block the conversion of xylitol to xylulose. Then, an NADPH regeneration system was added through heterologous expression of the Yarrowia lipolytica genes encoding 6-phosphate-gluconic acid dehydrogenase and 6-phosphate-glucose dehydrogenase. After process optimization, the engineered strain, C. tropicalis XZX-B4ZG, produced 97.10 g L− 1 xylitol in 120 h from 300 g L− 1 XML in a 5-L fermenter. The xylitol production rate was 0.82 g L− 1 h− 1 and the conversion rate was 92.40 %. Conclusions In conclusion, this study performed a combination of metabolic engineering and process optimizing in C. tropicalis to enhance xylitol production from XML. The use of C. tropicalis XZX-B4ZG, therefore, provided a convenient method to transform the industrial by-product XML into the useful material xylitol.

show abstract

Section: Discussionmentioning

confidence: 99%

Stepwise metabolic engineering of Candida tropicalis for efficient xylitol production from xylose mother liquor

et al. 2021

View full text Add to dashboard Cite

show abstract

“…For sequential disruption of the W. sorbophila POX genes, URA 3 pop-out cassette was constructed according to a previously published protocol [ 46 ]. The W. sorbophila URA3 fragment was inserted into the Bam HI/ Bgl II sites of pGEM-T easy vector, and the glu gene was inserted into the Bam HI/ Bgl II sites at both ends of URA3 , as a repeating sequence in the vector.…”

Section: Methodsmentioning

confidence: 99%

Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources

Lee

Han

Lee

et al. 2018

Biotechnol Biofuels

View full text Add to dashboard Cite

BackgroundAs a sustainable industrial process, the production of dicarboxylic acids (DCAs), used as precursors of polyamides, polyesters, perfumes, plasticizers, lubricants, and adhesives, from vegetable oil has continuously garnered interest. Although the yeast Candida tropicalis has been used as a host for DCA production, additional strains are continually investigated to meet productivity thresholds and industrial needs. In this regard, the yeast Wickerhamiella sorbophila, a potential candidate strain, has been screened. However, the lack of genetic and physiological information for this uncommon strain is an obstacle that merits further research. To overcome this limitation, we attempted to develop a method to facilitate genetic recombination in this strain and produce high amounts of DCAs from methyl laurate using engineered W. sorbophila.ResultsIn the current study, we first developed efficient genetic engineering tools for the industrial application of W. sorbophila. To increase homologous recombination (HR) efficiency during transformation, the cell cycle of the yeast was synchronized to the S/G2 phase using hydroxyurea. The HR efficiency at POX1 and POX2 loci increased from 56.3% and 41.7%, respectively, to 97.9% in both cases. The original HR efficiency at URA3 and ADE2 loci was nearly 0% during the early stationary and logarithmic phases of growth, and increased to 4.8% and 25.6%, respectively. We used the developed tools to construct W. sorbophila UHP4, in which β-oxidation was completely blocked. The strain produced 92.5 g/l of dodecanedioic acid (DDDA) from methyl laurate over 126 h in 5-l fed-batch fermentation, with a productivity of 0.83 g/l/h.ConclusionsWickerhamiella sorbophila UHP4 produced more DDDA methyl laurate than C. tropicalis. Hence, we demonstrated that W. sorbophila is a powerful microbial platform for vegetable oil-based DCA production. In addition, by using the developed genetic engineering tools, this emerging yeast could be used for the production of a variety of fatty acid derivatives, such as fatty alcohols, fatty aldehydes, and ω-hydroxy fatty acids.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1310-x) contains supplementary material, which is available to authorized users.

show abstract

“…The construction of the expression cassette was described previously [16]. To integrate the expression cassette into XYL3 locus of C. tropicalis K1 CoSTP2 LsaAraAp, the pXK4 plasmid was synthesized containing the construct of a pair of 60-bp homologous fragments of XYL3 (XK4F and XK4R) with the NruI sites at both ends and the BamHI site in the middle of XK4F and XK4R.…”

Section: Codon Optimization and Construction Of Expression Cassettementioning

confidence: 99%

“…However, C. tropicalis cannot assimilate l-arabinose, because the cells do not possess genes related to the l-arabinose metabolic pathway. Yoon et al (2011) expressed the three genes related to the bacterial l-arabinose metabolic pathway, araA (l-arabinose isomerase), araB (l-ribulokinase), and araD (l-ribulose-5-phosphate 4-epimerase), in C. tropicalis and constructed an l-arabinose-assimilating strain to reduce l-arabitol accumulation as a by-product during xylitol production. As a result, the recombinant cells assimilated l-arabinose without the accumulation of l-arabitol in the medium [16].…”

Section: Introductionmentioning

confidence: 99%

Conversion of l-arabinose to l-ribose by genetically engineered Candida tropicalis

Yeo

Cho

Kim

2021

Bioprocess Biosyst Eng

Self Cite

View full text Add to dashboard Cite

l -Ribose, a starting material for the synthesis of l -nucleoside, has attracted lots of attention since l -nucleoside is responsible for the antiviral activities of the racemic mixtures of nucleoside enantiomers. In this study, the l -ribulose-producing Candida tropicalis strain was engineered for the conversion of l -arabinose to l -ribose. For the construction of a uracil auxotroph, the URA3 gene was excised by homologous recombination. The expression cassette of codon-optimized l -ribose isomerase gene from Acinetobacter calcoaceticus DL-28 under the control of the GAPDH promoter was integrated to the uracil auxotroph. The resulting strain, K1 CoSTP2 LsaAraA AcLRI, was cultivated with the glucose/ l -arabinose mixture. At 45.5 h of fermentation, 6.0 g/L of l -ribose and 3.2 g/L of l -ribulose were produced from 30 g/L of l -arabinose. The proportion between l -ribose and l -ribulose was approximately 2:1 and the conversion yield of l -arabinose to l -ribose was about 20% ( w / w ). The l -ribose-producing yeast strain was successfully constructed for the first time and could convert l -arabinose to l -ribose in one-pot fermentation using the mixture of glucose and l -arabinose. Supplementary Information The online version contains supplementary material available at 10.1007/s00449-020-02506-2.

show abstract

l-Arabinose pathway engineering for arabitol-free xylitol production in Candida tropicalis

Cited by 19 publications

References 8 publications

Stepwise metabolic engineering of Candida tropicalis for efficient xylitol production from xylose mother liquor

Stepwise metabolic engineering of Candida tropicalis for efficient xylitol production from xylose mother liquor

Development of a promising microbial platform for the production of dicarboxylic acids from biorenewable resources

Conversion of l-arabinose to l-ribose by genetically engineered Candida tropicalis

Contact Info

Product

Resources

About