D-Galacturonic acid reduction by S. cerevisiae for L-galactonate production from extracted sugar beet press pulp hydrolysate

Wagner, Jacqueline; Schäfer, Dominik; Haimerl, C.; Harth, Simon; Oreb, Mislav; Benz, J. Philipp; Weuster‐Botz, Dirk

doi:10.1007/s00253-021-11433-5

Cited by 3 publications

(5 citation statements)

References 65 publications

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“…Ideally, one of the sugar monomers released from SBPP will be used as an educt to be transformed with the highest selectivity to a value-added product, and the other sugars will serve as co-substrates. For example, the released D-galacturonic acid in the hydrolysate may be used for further biotransformation to L-galactonate by recombinant yeast, as shown before [24,25]. In addition, L-arabinose may be converted to low calorie sweetener arabitol by Candida parapsilosis [62].…”

Section: Discussionmentioning

confidence: 99%

“…Complete conversion of single sugar monomers to higher-value products is thus inevitable for hydrolysates' economic utilization. As an example, the increase in D-GalA concentrations to 12 g L −1 shown here will improve the final concentration of the higher-value product L-galactonate, which can be produced by engineered Saccharomyces cerevisiae with high yields within a consecutive biotransformation process [24,25].…”

Section: Subsequent Hydrolysis Of Sbppmentioning

confidence: 92%

“…The hydrolysis of 90 g L −1 milled SBPP was studied in subsequent submerged batch processes with sterile filtered enzyme supernatants, resulting in the release of 8.8 g L −1 of D-GalA, which corresponded to a D-GalA yield of ~36.4% [23]. Using engineered Saccharomyces cerevisiae, the resulting D-GalA solution may then be reduced within a consecutive biotransformation process with a product selectivity of 97% to the higher value product L-galactonate (L-GalOA) [24,25], which was shown to have promise, for example, as an alternative leavening agent in baking applications [26].…”

Section: Introductionmentioning

confidence: 99%

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Enzymatic One-Pot Hydrolysis of Extracted Sugar Beet Press Pulp after Solid-State Fermentation with an Engineered Aspergillus niger Strain

Knesebeck,

Schäfer,

Schmitz

et al. 2023

Fermentation

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Extracted sugar beet press pulp (SBPP) is a promising agricultural residue for saccharification and further bioconversion. Combining solid-state fermentation of SBPP with engineered Aspergillus niger for enzyme production followed by hydrolysis of additionally added SBPP in the same bioreactor was studied to produce a sugar solution (hydrolysate) in a one-pot process. The initial aerobic solid-state fermentations were carried out in duplicate on non-milled, wet SBPP (moisture content of 72% (w/v)) with an A. niger strain engineered for constitutive pectinase production for 96 h, and this resulted in polygalacturonase activities of up to 256 U mL−1 in the wet media. Afterwards, water was added to the bioreactor, and the remaining solids were suspended by stirring to dissolve the hydrolytic enzymes. Metabolic activities of A. niger were inactivated by a N2-atmosphere and by increasing the temperature to 50 °C. High solid loads of milled SBPP were added to the stirred-tank reactor with a delay of 24 h to enable sugar yield calculations based on the compositional analysis of the SBPP used. The resulting final sugar concentrations of the hydrolysate after 166 h were 17 g L−1 d-glucose, 18.8 g L−1 l-arabinose, and 12.5 g L−1 d-galacturonic acid, corresponding to sugar yields of 98% d-glucose, 86% l-arabinose, and 50% d-galacturonic acid, respectively. Including the other sugars released during enzymatic hydrolysis in the one-pot process (d-xylose, d-mannose, d-galactose), a total sugar concentration of 54.8 g L−1 was achieved in the hydrolysate. The one-pot process combining hydrolytic enzyme production in solid-state fermentation with high solid loads during enzymatic hydrolysis of the milled SBPP reduces hydrolytic process costs by replacing chemical pre-treatments, enabling the in situ production of SBPP-adapted hydrolytic enzymes, as well as avoiding intermediate enzyme extraction and preparation steps.

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Section: Discussionmentioning

confidence: 99%

Section: Subsequent Hydrolysis Of Sbppmentioning

confidence: 92%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Enzymatic One-Pot Hydrolysis of Extracted Sugar Beet Press Pulp after Solid-State Fermentation with an Engineered Aspergillus niger Strain

Knesebeck,

Schäfer,

Schmitz

et al. 2023

Fermentation

View full text Add to dashboard Cite

show abstract

“…As detailed in the introduction, it is obvious to equip baker's yeast with substrate consumption pathways that allow ethanol production from pectin-rich biomass including D-GalUA. However, baker's yeast has also been considered for biotransformations of D-GalUA to meso-galactarate [27] and L-GalA [32,61,64]. Galactarate can be further converted to adipic acid [27,65], and L-GalA is a precursor of L-ascorbic acid (vitamin C) synthesis [66].…”

Section: Discussionmentioning

confidence: 99%

Identification of the Aldo-Keto Reductase Responsible for d-Galacturonic Acid Conversion to l-Galactonate in Saccharomyces cerevisiae

Rippert¹,

Linguardo²,

Perpelea³

et al. 2021

JoF

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d-galacturonic acid (d-GalUA) is the main constituent of pectin, a complex polysaccharide abundant in several agro-industrial by-products such as sugar beet pulp or citrus peel. During several attempts to valorise d-GalUA by engineering the popular cell factory Saccharomyces cerevisiae, it became obvious that d-GalUA is, to a certain degree, converted to l-galactonate (l-GalA) by an endogenous enzymatic activity. The goal of the current work was to clarify the identity of the responsible enzyme(s). A protein homology search identified three NADPH-dependent unspecific aldo-keto reductases in baker’s yeast (encoded by GCY1, YPR1 and GRE3) that show sequence similarities to known d-GalUA reductases from filamentous fungi. Characterization of the respective deletion mutants and an in vitro enzyme assay with a Gcy1 overproducing strain verified that Gcy1 is mainly responsible for the detectable reduction of d-GalUA to l-GalA.

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“…SP02 coding an arsenal of enzymes involved in depolymerization of pectins such as endo-polygalacturonase (EC:3.2.1.15), exo-polygalacturonase (EC:3.2.1.67), rhamnogalacturonan endolyase (EC:4.2.2.23), α-L-rhamnosidase (EC:3.2.1.40), pectate lyase (EC:4.2.2.2), pectin esterase (EC:3.1.1.11), and rhamnogalacturonan acetylesterase (EC:3.1.1.86). Endo-and exo-polygalacturonases act by cleaving linkages of homogalacturonan to release D-galacturonic acid [93]. Rhamnogalacturonan endolyase and α-Lrhamnosidase are involved in depolymerization of rhamnogalacturonan [70].…”

Section: Other Degrading Enzymesmentioning

confidence: 99%

Genomic Studies of White-Rot Fungus Cerrena unicolor SP02 Provide Insights into Food Safety Value-Added Utilization of Non-Food Lignocellulosic Biomass

Zhang

Shah

Mohamed

et al. 2021

JoF

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Cerrena unicolor is an ecologically and biotechnologically important wood-degrading basidiomycete with high lignocellulose degrading ability. Biological and genetic investigations are limited in the Cerrena genus and, thus, hinder genetic modification and commercial use. The aim of the present study was to provide a global understanding through genomic and experimental research about lignocellulosic biomass utilization by Cerrena unicolor. In this study, we reported the genome sequence of C. unicolor SP02 by using the Illumina and PacBio 20 platforms to obtain trustworthy assembly and annotation. This is the combinational 2nd and 3rd genome sequencing and assembly of C. unicolor species. The generated genome was 42.79 Mb in size with an N50 contig size of 2.48 Mb, a G + C content of 47.43%, and encoding of 12,277 predicted genes. The genes encoding various lignocellulolytic enzymes including laccase, lignin peroxidase, manganese peroxidase, cytochromes P450, cellulase, xylanase, α-amylase, and pectinase involved in the degradation of lignin, cellulose, xylan, starch, pectin, and chitin that showed the C. unicolor SP02 potentially have a wide range of applications in lignocellulosic biomass conversion. Genome-scale metabolic analysis opened up a valuable resource for a better understanding of carbohydrate-active enzymes (CAZymes) and oxidoreductases that provide insights into the genetic basis and molecular mechanisms for lignocellulosic degradation. The C. unicolor SP02 model can be used for the development of efficient microbial cell factories in lignocellulosic industries. The understanding of the genetic material of C. unicolor SP02 coding for the lignocellulolytic enzymes will significantly benefit us in genetic manipulation, site-directed mutagenesis, and industrial biotechnology.

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D-Galacturonic acid reduction by S. cerevisiae for L-galactonate production from extracted sugar beet press pulp hydrolysate

Cited by 3 publications

References 65 publications

Enzymatic One-Pot Hydrolysis of Extracted Sugar Beet Press Pulp after Solid-State Fermentation with an Engineered Aspergillus niger Strain

Enzymatic One-Pot Hydrolysis of Extracted Sugar Beet Press Pulp after Solid-State Fermentation with an Engineered Aspergillus niger Strain

Identification of the Aldo-Keto Reductase Responsible for d-Galacturonic Acid Conversion to l-Galactonate in Saccharomyces cerevisiae

Genomic Studies of White-Rot Fungus Cerrena unicolor SP02 Provide Insights into Food Safety Value-Added Utilization of Non-Food Lignocellulosic Biomass

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