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Background: Xylitol is a five-carbon sugar alcohol that has numerous beneficial health properties. It has almost the same sweetness as sucrose but has lower energy value compared to the sucrose. Metabolism of xylitol is insulin independent and thus it is an ideal sweetener for diabetics. It is widely used in food products, oral and personal care, and animal nutrition as well. Here we present a two-stage strategy to produce bio-xylitol from D-xylose using a recombinant Pichia pastoris expressing a heterologous xylose reductase gene. The recombinant P. pastoris cells were first generated by a low-cost, standard procedure. The cells were then used as a catalyst to make the bio-xylitol from D-xylose.
Results: P. pastoris expressing XYL1 from P. stipitis and gdh from B. subtilis demonstrated that the biotransformation was very efficient with as high as 80% (w/w) conversion within two hours. The whole cells could be re-used for multiple rounds of catalysis without loss of activity. Also, the cells could directly transform D-xylose in a non-detoxified hemicelluloses hydrolysate to xylitol at 70% (w/w) yield.
Conclusions: We demonstrated here that the recombinant P. pastoris expressing xylose reductase could transform D-xylose, either in pure form or in crude hemicelluloses hydrolysate, to bio-xylitol very efficiently. This biocatalytic reaction happened without the external addition of any NAD(P)H, NAD(P)+, and auxiliary substrate as an electron donor. Our experimental design & findings reported here are not limited to the conversion of D-xylose to xylitol only but can be used with other many oxidoreductase reactions also, such as ketone reductases/alcohol dehydrogenases and amino acid dehydrogenases, which are widely used for the synthesis of high-value chemicals and pharmaceutical intermediates.
Keywords
Xylose reductase, glucose dehydrogenase, whole cells, Pichia, xylose, and xylitol
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View the published article at Microbial Cell Factories.
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Review #2 received
Received 28 Jan, 2021
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On 28 Jan, 2021
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On 26 Jan, 2021
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Received 19 Jan, 2021
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On 16 Jan, 2021
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On 15 Jan, 2021
Reviewers invited
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On 15 Jan, 2021
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On 15 Jan, 2021
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Review #2 received
Received 30 Dec, 2020
Editorial decision: Minor Revision
On 30 Dec, 2020
Reviewer #2 agreed
On 29 Dec, 2020
Review #1 received
Received 03 Dec, 2020
Reviewer #1 agreed
On 26 Nov, 2020
Reviewers invited
Invitations sent on 21 Nov, 2020
Editor assigned
On 20 Nov, 2020
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On 20 Nov, 2020
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On 20 Nov, 2020
Version (no preprint)
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On 29 Aug, 2020
Review #2 received
Received 28 Aug, 2020
Reviewer #2 agreed
On 21 Aug, 2020
Review #1 received
Received 08 Aug, 2020
Reviewer #1 agreed
On 07 Aug, 2020
Reviewers invited
Invitations sent on 05 Aug, 2020
Editor assigned
On 05 Aug, 2020
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On 04 Aug, 2020
Editor invited
On 04 Aug, 2020
This version was not preprinted
Version 1
Posted 01 May, 2020
No comments provided
Review #3 received
Received 06 Jul, 2020
Reviewer #3 agreed
On 22 Jun, 2020
Review #2 received
Received 22 May, 2020
Reviewer #2 agreed
On 17 May, 2020
Reviewer #1 agreed
On 27 Apr, 2020
Editor assigned
On 26 Apr, 2020
Reviewers invited
Invitations sent on 26 Apr, 2020
Submission checks complete
On 25 Apr, 2020
Editor invited
On 25 Apr, 2020
First submitted
On 23 Apr, 2020
Metrics
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PDF Downloads: ...
HTML Views: ...
Subject Areas
Applied & Industrial Microbiology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Microbial Cell Factories.
Background: Xylitol is a five-carbon sugar alcohol that has numerous beneficial health properties. It has almost the same sweetness as sucrose but has lower energy value compared to the sucrose. Metabolism of xylitol is insulin independent and thus it is an ideal sweetener for diabetics. It is widely used in food products, oral and personal care, and animal nutrition as well. Here we present a two-stage strategy to produce bio-xylitol from D-xylose using a recombinant Pichia pastoris expressing a heterologous xylose reductase gene. The recombinant P. pastoris cells were first generated by a low-cost, standard procedure. The cells were then used as a catalyst to make the bio-xylitol from D-xylose.
Results: P. pastoris expressing XYL1 from P. stipitis and gdh from B. subtilis demonstrated that the biotransformation was very efficient with as high as 80% (w/w) conversion within two hours. The whole cells could be re-used for multiple rounds of catalysis without loss of activity. Also, the cells could directly transform D-xylose in a non-detoxified hemicelluloses hydrolysate to xylitol at 70% (w/w) yield.
Conclusions: We demonstrated here that the recombinant P. pastoris expressing xylose reductase could transform D-xylose, either in pure form or in crude hemicelluloses hydrolysate, to bio-xylitol very efficiently. This biocatalytic reaction happened without the external addition of any NAD(P)H, NAD(P)+, and auxiliary substrate as an electron donor. Our experimental design & findings reported here are not limited to the conversion of D-xylose to xylitol only but can be used with other many oxidoreductase reactions also, such as ketone reductases/alcohol dehydrogenases and amino acid dehydrogenases, which are widely used for the synthesis of high-value chemicals and pharmaceutical intermediates.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
The full text of this article is available to read as a PDF.
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