Formation of xylitol and xylitol-5-phosphate and its impact on growth of d-xylose-utilizing Corynebacterium glutamicum strains

Radek, Andreas; Müller, Moritz-Fabian; Gätgens, Jochem; Eggeling, Lothar; Krumbach, Karin; Marienhagen, Jan; Noack, Stephan

doi:10.1016/j.jbiotec.2016.06.009

Cited by 15 publications

(9 citation statements)

References 17 publications

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“…Interestingly, SL-1 harboring the empty vector with 2% (w/v) glucose and 2% (w/v) xylose also consumed 2.2 g/L xylose (14.9 mM) and produced 1.5 g/L (10.13 mM) xylitol. Because the engineered C. glutamicum , exhibiting the Weimberg pathway, has been reported to accumulate small amounts of xylitol when grown on a mixture of glucose and xylose, 25 we assumed that C. glutamicum WT, under conditions in which xylose metabolism is not efficient, could enable the oxidation of xylose to xylitol.…”

Section: Resultsmentioning

confidence: 99%

Bioconversion of Xylose to Ethylene Glycol and Glycolate in Engineered Corynebacterium glutamicum

2019

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The biological production of two-carbon compounds (ethylene glycol (EG) and glycolate) has been studied for the sustainable supply of the compounds to the polymer, cosmetic, textile, and medical industries. Here, we demonstrated the bioconversion of xylose to either ethylene glycol (EG) or glycolate using engineered Corynebacterium glutamicum, a well-known industrial amino acid producer. A synthetic ribulose 1-phosphate (Ru1P) pathway involving heterologous d-tagatose 3-epimerase and l-fuculose kinase/aldolase reactions was introduced in C. glutamicum. Subsequently, heterologous expression of Escherichia coli YqhD reductase with the synthetic Ru1P pathway led to ethylene glycol production from xylose. Additional pathway engineering in C. glutamicum by mutating ald, which encodes an aldehyde dehydrogenase, abolished the by-product formation of glycolate during xylose conversion to EG at a yield of 0.75 mol per mol. In addition, the bioconversion of xylose to glycolate was achieved, and the almost maximum molar yield was 0.99 mol per mol xylose in C. glutamicum via the Ru1P pathway. Thus, the synthetic Ru1P pathway in C. glutamicum led bioconversion of xylose to either ethylene glycol or glycolate with high molar yields.

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Section: Resultsmentioning

confidence: 99%

Bioconversion of Xylose to Ethylene Glycol and Glycolate in Engineered Corynebacterium glutamicum

2019

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“…Kawaguchi et al ., 2008; Radek et al ., 2014). In contrast to the isomerase pathway, the Weimberg pathway enables a more carbon efficient utilization of d ‐xylose and allows a parallel consumption of d ‐xylose and d ‐glucose in C. glutamicum (Radek et al ., 2014, 2016). However, the maximal net generated biomass (4.7 ± 0.4 g CDW L −1 ) was doubled with respect to sole d ‐glucose (2.2 ± 0.1 g CDW L −1 ), and the higher cell density allowed a full consumption of d ‐xylose and 80% of l ‐arabinose within the given cultivation time (cf.…”

Section: Resultsmentioning

confidence: 99%

Harnessing novel chromosomal integration loci to utilize an organosolv‐derived hemicellulose fraction for isobutanol production with engineered Corynebacterium glutamicum

Lange

Müller

Blombach

2017

Microbial Biotechnology

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SummaryA successful bioeconomy depends on the manifestation of biorefineries that entirely convert renewable resources to valuable products and energies. Here, the poorly exploited hemicellulose fraction (HF) from beech wood organosolv processing was applied for isobutanol production with Corynebacterium glutamicum. To enable growth of C. glutamicum on HF, we integrated genes required for d‐xylose and l‐arabinose metabolization into two of 16 systematically identified and novel chromosomal integration loci. Under aerobic conditions, this engineered strain CArXy reached growth rates up to 0.34 ± 0.02 h−1 on HF. Based on CArXy, we developed the isobutanol producer strain CIsArXy, which additionally (over)expresses genes of the native l‐valine biosynthetic and the heterologous Ehrlich pathway. CIsArXy produced 7.2 ± 0.2 mM (0.53 ± 0.02 g L−1) isobutanol on HF at a carbon molar yield of 0.31 ± 0.02 C‐mol isobutanol per C‐mol substrate (d‐xylose + l‐arabinose) in an anaerobic zero‐growth production process.

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“…LC–MS/MS analysis was performed on a AB ScieX Triple ToF 6600 connected with an Agilent 1260 HPLC as described in literature [36] from the last sampling time points of the bioreactor cultivations with CaCl 2 . The secretome data is available at the PRIDE [37] repository with the identifier PXD013751.…”

Section: Methodsmentioning

confidence: 99%

A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping

Jansen

Beuck

Moch

et al. 2019

Fungal Biol Biotechnol

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Background Filamentously growing microorganisms offer unique advantages for biotechnological processes, such as extraordinary secretion capacities, going along with multiple obstacles due to their complex morphology. However, limited experimental throughput in bioprocess development still hampers taking advantage of their full potential. Miniaturization and automation are powerful tools to accelerate bioprocess development, but so far the application of such technologies has mainly been focused on non-filamentous systems. During cultivation, filamentous fungi can undergo remarkable morphological changes, creating challenging cultivation conditions. Depending on the process and product, only one specific state of morphology may be advantageous to achieve e.g. optimal productivity or yield. Different approaches to control morphology have been investigated, such as microparticle enhanced cultivation. However, the addition of solid microparticles impedes the optical measurements typically used by microbioreactor systems and thus alternatives are needed. Results Aspergillus giganteus IfGB 0902 was used as a model system to develop a time-efficient and robust workflow allowing microscale cultivation with increased throughput. The effect of microtiter plate geometry, shaking frequency and medium additives (talc and calcium chloride) on homogeneity of culture morphology as well as reproducibility were analyzed via online biomass measurement, microscopic imaging and cell dry weight. While addition of talc severely affected online measurements, 2% (w v −1 ) calcium chloride was successfully applied to obtain a highly reproducible growth behavior with homogenous morphology. Furthermore, the influence of small amounts of complex components was investigated for the applied model strain. By correlation to cell dry weight, it could be shown that optical measurements are a suitable signal for biomass concentration. However, each correlation is only applicable for a specific set of cultivation parameters. These optimized conditions were used in micro as well as lab-scale bioreactor cultivation in order to verify the reproducibility and scalability of the setup. Conclusion A robust workflow for A. giganteus was developed, allowing for reproducible microscale cultivation with online monitoring, where calcium chloride is an useful alternative to microparticle enhanced cultivation in order to control the morphology. Independent of the cultivation volume, comparable phenotypes were observed in microtiter plates and in lab-scale bioreactor. Electronic supplementary material The online version of this article (10.1186/s40694-019-0073-x) contains supplementary material, which is available to authorized users.

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Formation of xylitol and xylitol-5-phosphate and its impact on growth of d-xylose-utilizing Corynebacterium glutamicum strains

Cited by 15 publications

References 17 publications

Bioconversion of Xylose to Ethylene Glycol and Glycolate in Engineered Corynebacterium glutamicum

Bioconversion of Xylose to Ethylene Glycol and Glycolate in Engineered Corynebacterium glutamicum

Harnessing novel chromosomal integration loci to utilize an organosolv‐derived hemicellulose fraction for isobutanol production with engineered Corynebacterium glutamicum

A closer look at Aspergillus: online monitoring via scattered light enables reproducible phenotyping

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