Enhancement of xylitol production in Candida tropicalis by co-expression of two genes involved in pentose phosphate pathway

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

doi:10.1007/s00449-011-0641-9

Cited by 44 publications

(24 citation statements)

References 22 publications

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“…The xylE activity assay (20) confirmed their much stronger activities than the control promoter, ermE* p , (Figure 1b), which is the mutated variant of the promoter of the erythromycin resistance gene from Saccharopolyspora erythraea , and is believed to be one of the strongest constitutive promoters in Streptomycetes (21, 22). The strong activities of gapdh p and the promoters of various translation elongation factors have also been observed in many other microorganisms, such as fungi, bacteria, micro-algae, and protozoa (23–29). Encouraged by the strong activities of gapdh p and rpsL p , we decided to examine their equivalents from other species.…”

Section: Resultsmentioning

confidence: 83%

Refactoring the Silent Spectinabilin Gene Cluster Using a Plug-and-Play Scaffold

et al. 2013

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Natural products (secondary metabolites) are a rich source of compounds with important biological activities. Eliciting pathway expression is always challenging but extremely important in natural product discovery because individual pathway is tightly controlled through unique regulation mechanism and hence often remains silent in the routine culturing conditions. To overcome the drawback of the traditional approaches that lack general applicability, we developed a simple synthetic biology approach that decouples pathway expression from complex native regulations. Briefly, the entire silent biosynthetic pathway is refactored using a plug-and-play scaffold and a set of heterologous promoters that are functional in a heterologous host under the target culturing condition. Using this strategy, we successfully awakened the silent spectinabilin pathway from Streptomyces orinoci. This strategy bypasses the traditional laborious processes to elicit pathway expression and represents a new platform for discovering novel natural products.

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Section: Resultsmentioning

confidence: 83%

Refactoring the Silent Spectinabilin Gene Cluster Using a Plug-and-Play Scaffold

et al. 2013

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“…In Candida utilis , for example, two enzymes involved in the pentose phosphate pathway (PPP) including glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase are responsible for the production of the required NADPH [17]. A study by Ahmad et al [5] showed that overexpression of these two enzymes led to an increase in xylitol production in Candida tropicalis .…”

Section: Discussionmentioning

confidence: 99%

“…Studies have been carried out for xylitol production in yeast, especially in Candida species and Saccharomyces cerevisiae [5–7]. However, fewer studies have been completed on xylitol production in T. reesei .…”

Section: Introductionmentioning

confidence: 99%

Overexpression of D-Xylose Reductase (xyl1) Gene and Antisense Inhibition of D-Xylulokinase (xyiH) Gene Increase Xylitol Production inTrichoderma reesei

Yang

Dashtban

Kepka

et al. 2014

BioMed Research International

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T. reesei is an efficient cellulase producer and biomass degrader. To improve xylitol production in Trichoderma reesei strains by genetic engineering, two approaches were used in this study. First, the presumptive D-xylulokinase gene in T. reesei (xyiH), which has high homology to known fungi D-xylulokinase genes, was silenced by transformation of T. reesei QM9414 strain with an antisense construct to create strain S6-2-2. The expression of the xyiH gene in the transformed strain S6-2-2 decreased at the mRNA level, and D-xylulokinase activity decreased after 48 h of incubation. This led to an increase in xylitol production from undetectable levels in wild-type T. reesei QM9414 to 8.6 mM in S6-2-2. The T. reesei Δxdh is a xylose dehydrogenase knockout strain with increased xylitol production compared to the wild-type T. reesei QM9414 (22.8 mM versus undetectable). The copy number of the xylose reductase gene (xyl1) in T. reesei Δxdh strain was increased by genetic engineering to create a new strain Δ9-5-1. The Δ9-5-1 strain showed a higher xyl1 expression and a higher yield of xylose reductase, and xylitol production was increased from 22.8 mM to 24.8 mM. Two novel strains S6-2-2 and Δ9-5-1 are capable of producing higher yields of xylitol. T. reesei has great potential in the industrial production of xylitol.

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“…Compared to the control promoter, much stronger activities of gapdh p (SG) and rpsL p (SG) should be observed. The strong activities of gapdh p and the promoters of various translation elongation factors have also been observed in many other microorganisms, such as fungi, bacteria, micro-algae, and protozoa (Krasny et al, 2000;Suarez et al, 2006;Ahn et al, 2007;Shao et al, 2009;Hong et al, 2010;Ahmad et al, 2012;Jia et al, 2012). Cloning additional promoters 24.…”

Section: Now the Synthesized Cdna Is Ready For Real-time Pcrmentioning

confidence: 94%

Manipulating Natural Product Biosynthetic Pathways via DNA Assembler

Shao

Zhao

2014

CP Chemical Biology

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DNA assembler is an efficient synthetic biology method for constructing and manipulating biochemical pathways. The rapidly increasing number of sequenced genomes provides a rich source for discovery of gene clusters involved in synthesizing new natural products. However, both discovery and economical production are hampered by our limited knowledge in manipulating most organisms and the corresponding pathways. By taking advantage of yeast in vivo homologous recombination, DNA assembler synthesizes an entire expression vector containing the target biosynthetic pathway and the genetic elements needed for DNA maintenance and replication. Here we use the spectinabilin clusters originated from two hosts as examples to illustrate the guidelines of using DNA assembler for cluster characterization and silent cluster activation. Such strategies offer unprecedented versatility in cluster manipulation, bypass the traditional laborious strategies to elicit pathway expression, and provide a new platform for de novo cluster assembly and genome mining for discovering new natural products. Curr. Protoc. Chem. Biol. 6:65‐100 © 2014 by John Wiley & Sons, Inc.

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Enhancement of xylitol production in Candida tropicalis by co-expression of two genes involved in pentose phosphate pathway

Cited by 44 publications

References 22 publications

Refactoring the Silent Spectinabilin Gene Cluster Using a Plug-and-Play Scaffold

Refactoring the Silent Spectinabilin Gene Cluster Using a Plug-and-Play Scaffold

Overexpression of D-Xylose Reductase (xyl1) Gene and Antisense Inhibition of D-Xylulokinase (xyiH) Gene Increase Xylitol Production inTrichoderma reesei

Manipulating Natural Product Biosynthetic Pathways via DNA Assembler

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