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Induction of D-xylose uptake and expression of NAD(P)H-linked xylose reductase and NADP + -linked xylitol dehydrogenase in the oleaginous microalga Chlorella sorokiniana.

Zheng Y, Yu X, Li T, Xiong X, Chen S - Biotechnol Biofuels (2014)

Bottom Line: The uptake of D-xylose activated the related metabolic pathway, and the activities of a NAD(P)H-linked xylose reductase (XR) and a unique NADP(+)-linked xylitol dehydrogenase (XDH) were detected in C. sorokiniana.The uptake of D-xylose subsequently activated the expression of key catalytic enzymes that enabled D-xylose entering central metabolism.Results of this research are useful to better understand the D-xylose metabolic pathway in the microalga C. sorokiniana and provide a target for genetic engineering to improve D-xylose utilization for microalgal lipid production.

View Article: PubMed Central - PubMed

Affiliation: LJ Smith 258, Biological Systems Engineering, Washington State University, Pullman, WA 99164 USA.

ABSTRACT

Background: The heterotrophic and mixotrophic culture of oleaginous microalgae is a promising process to produce biofuel feedstock due to the advantage of fast growth. Various organic carbons have been explored for this application. However, despite being one of the most abundant and economical sugar resources in nature, D-xylose has never been demonstrated as a carbon source for wild-type microalgae. The purpose of the present work was to identify the feasibility of D-xylose utilization by the oleaginous microalga Chlorella sorokiniana.

Results: The sugar uptake kinetic analysis was performed with (14)C-labeled sugars and the data showed that the D-glucose induced algal cells (the alga was heterotrophically grown on D-glucose and then harvested as D-glucose induced cells) exhibited a remarkably increased D-xylose uptake rate. The maximum D-xylose transport rate was 3.8 nmol min(-1) mg(-1) dry cell weight (DCW) with K m value of 6.8 mM. D-xylose uptake was suppressed in the presence of D-glucose, D-galactose and D-fructose but not L-arabinose and D-ribose. The uptake of D-xylose activated the related metabolic pathway, and the activities of a NAD(P)H-linked xylose reductase (XR) and a unique NADP(+)-linked xylitol dehydrogenase (XDH) were detected in C. sorokiniana. Compared with the culture in the dark, the consumption of D-xylose increased 2 fold under light but decreased to the same level with addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), indicating that extra chemical energy from the light-dependent reaction contributed the catabolism of D-xylose for C. sorokiniana.

Conclusions: An inducible D-xylose transportation system and a related metabolic pathway were discovered for microalga for the first time. The transportation of D-xylose across the cell membrane of C. sorokiniana could be realized by an inducible hexose symporter. The uptake of D-xylose subsequently activated the expression of key catalytic enzymes that enabled D-xylose entering central metabolism. Results of this research are useful to better understand the D-xylose metabolic pathway in the microalga C. sorokiniana and provide a target for genetic engineering to improve D-xylose utilization for microalgal lipid production.

No MeSH data available.


D-xylose uptake kinetics of induced (black circle) and non-induced (white triangle)C. sorokinianausing two different models. Model I (single-carrier, solid line) and Model II (two-carrier, dash line).
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Fig1: D-xylose uptake kinetics of induced (black circle) and non-induced (white triangle)C. sorokinianausing two different models. Model I (single-carrier, solid line) and Model II (two-carrier, dash line).

Mentions: The consumption of D-xylose by C. sorokiniana was observed in the D-glucose induced algal cells (the alga was grown on D-glucose heterotrophically and then harvested as D-glucose induced cells). The kinetic parameters of two different models for D-xylose transport in C. sorokiniana are listed in Table 1. For induced algal cells, there was no significant difference of variances between these two models. As seen in Figure 1, the two-carrier model fitted the experimental data better than the single-carrier model. However, the Vmax2 and Km2 of the two-carrier model were extremely high, which was unreasonable for representing the transportation kinetics. Thus, the single-carrier model was selected to represent the D-xylose transport for induced C. sorokiniana. Non-linear regression analysis showed that the maximum D-xylose transport rate was 3.8 nmol min−1 mg−1 dry cell weight (DCW)with Km value of 6.8 mM.Table 1


Induction of D-xylose uptake and expression of NAD(P)H-linked xylose reductase and NADP + -linked xylitol dehydrogenase in the oleaginous microalga Chlorella sorokiniana.

Zheng Y, Yu X, Li T, Xiong X, Chen S - Biotechnol Biofuels (2014)

D-xylose uptake kinetics of induced (black circle) and non-induced (white triangle)C. sorokinianausing two different models. Model I (single-carrier, solid line) and Model II (two-carrier, dash line).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4195881&req=5

Fig1: D-xylose uptake kinetics of induced (black circle) and non-induced (white triangle)C. sorokinianausing two different models. Model I (single-carrier, solid line) and Model II (two-carrier, dash line).
Mentions: The consumption of D-xylose by C. sorokiniana was observed in the D-glucose induced algal cells (the alga was grown on D-glucose heterotrophically and then harvested as D-glucose induced cells). The kinetic parameters of two different models for D-xylose transport in C. sorokiniana are listed in Table 1. For induced algal cells, there was no significant difference of variances between these two models. As seen in Figure 1, the two-carrier model fitted the experimental data better than the single-carrier model. However, the Vmax2 and Km2 of the two-carrier model were extremely high, which was unreasonable for representing the transportation kinetics. Thus, the single-carrier model was selected to represent the D-xylose transport for induced C. sorokiniana. Non-linear regression analysis showed that the maximum D-xylose transport rate was 3.8 nmol min−1 mg−1 dry cell weight (DCW)with Km value of 6.8 mM.Table 1

Bottom Line: The uptake of D-xylose activated the related metabolic pathway, and the activities of a NAD(P)H-linked xylose reductase (XR) and a unique NADP(+)-linked xylitol dehydrogenase (XDH) were detected in C. sorokiniana.The uptake of D-xylose subsequently activated the expression of key catalytic enzymes that enabled D-xylose entering central metabolism.Results of this research are useful to better understand the D-xylose metabolic pathway in the microalga C. sorokiniana and provide a target for genetic engineering to improve D-xylose utilization for microalgal lipid production.

View Article: PubMed Central - PubMed

Affiliation: LJ Smith 258, Biological Systems Engineering, Washington State University, Pullman, WA 99164 USA.

ABSTRACT

Background: The heterotrophic and mixotrophic culture of oleaginous microalgae is a promising process to produce biofuel feedstock due to the advantage of fast growth. Various organic carbons have been explored for this application. However, despite being one of the most abundant and economical sugar resources in nature, D-xylose has never been demonstrated as a carbon source for wild-type microalgae. The purpose of the present work was to identify the feasibility of D-xylose utilization by the oleaginous microalga Chlorella sorokiniana.

Results: The sugar uptake kinetic analysis was performed with (14)C-labeled sugars and the data showed that the D-glucose induced algal cells (the alga was heterotrophically grown on D-glucose and then harvested as D-glucose induced cells) exhibited a remarkably increased D-xylose uptake rate. The maximum D-xylose transport rate was 3.8 nmol min(-1) mg(-1) dry cell weight (DCW) with K m value of 6.8 mM. D-xylose uptake was suppressed in the presence of D-glucose, D-galactose and D-fructose but not L-arabinose and D-ribose. The uptake of D-xylose activated the related metabolic pathway, and the activities of a NAD(P)H-linked xylose reductase (XR) and a unique NADP(+)-linked xylitol dehydrogenase (XDH) were detected in C. sorokiniana. Compared with the culture in the dark, the consumption of D-xylose increased 2 fold under light but decreased to the same level with addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), indicating that extra chemical energy from the light-dependent reaction contributed the catabolism of D-xylose for C. sorokiniana.

Conclusions: An inducible D-xylose transportation system and a related metabolic pathway were discovered for microalga for the first time. The transportation of D-xylose across the cell membrane of C. sorokiniana could be realized by an inducible hexose symporter. The uptake of D-xylose subsequently activated the expression of key catalytic enzymes that enabled D-xylose entering central metabolism. Results of this research are useful to better understand the D-xylose metabolic pathway in the microalga C. sorokiniana and provide a target for genetic engineering to improve D-xylose utilization for microalgal lipid production.

No MeSH data available.