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Bioprocess and Biosystems Engineering Jun 2021L-Ribose, a starting material for the synthesis of L-nucleoside, has attracted lots of attention since L-nucleoside is responsible for the antiviral activities of the...
L-Ribose, a starting material for the synthesis of L-nucleoside, has attracted lots of attention since L-nucleoside is responsible for the antiviral activities of the racemic mixtures of nucleoside enantiomers. In this study, the L-ribulose-producing Candida tropicalis strain was engineered for the conversion of L-arabinose to L-ribose. For the construction of a uracil auxotroph, the URA3 gene was excised by homologous recombination. The expression cassette of codon-optimized L-ribose isomerase gene from Acinetobacter calcoaceticus DL-28 under the control of the GAPDH promoter was integrated to the uracil auxotroph. The resulting strain, K1 CoSTP2 LsaAraA AcLRI, was cultivated with the glucose/L-arabinose mixture. At 45.5 h of fermentation, 6.0 g/L of L-ribose and 3.2 g/L of L-ribulose were produced from 30 g/L of L-arabinose. The proportion between L-ribose and L-ribulose was approximately 2:1 and the conversion yield of L-arabinose to L-ribose was about 20% (w/w). The L-ribose-producing yeast strain was successfully constructed for the first time and could convert L-arabinose to L-ribose in one-pot fermentation using the mixture of glucose and L-arabinose.
Topics: Arabinose; Candida tropicalis; Microorganisms, Genetically-Modified; Ribose
PubMed: 33559750
DOI: 10.1007/s00449-020-02506-2 -
Applied Microbiology and Biotechnology Jun 2022Rhizobium sp. RM solubilized tri-calcium phosphate (TCP: 324-463 µg ml) and rock phosphate (RP: 36-46.58 µg ml) in the presence of common rhizospheric...
Rhizobium sp. RM solubilized tri-calcium phosphate (TCP: 324-463 µg ml) and rock phosphate (RP: 36-46.58 µg ml) in the presence of common rhizospheric sugars-glucose, arabinose, xylose and their combinations. Fructose, though did not support RP solubilization individually, surprisingly solubilized significantly higher phosphate when combined with aldoses. The highest TCP (644 µg ml) and RP (75 µg ml) solubilization was achieved in fructose + glucose combination. Presence of gluconate, malate and oxalate in culture supernatant indicated functioning of periplasmic glucose oxidation, the non-phosphorylative arabinose dehydrogenase pathway and the tricarboxylate (TCA) cycle, respectively. Aldoses, when present together, were co-utilized (monoauxic growth) however, when added with fructose, prevented the uptake of fructose yielding a typical diauxic growth. This presented an unusual sequential utilization of aldoses over a ketose (fructose) in strain RM. The prevention of fructose uptake by aldoses was investigated through real-time expression of key genes coding fructose transport proteins and initial enzymes of sugar metabolism. Fructose was actively transported via fructose-specific ABC transporters as suggested by upregulation of frcB and frcC only in fructose and fructose growth phases of fructose + aldose combinations. The probable route of initial fructose metabolism involved either fructokinase and/or xylose isomerase, as confirmed by enzyme activities. The upregulation of hfq and hprK genes only in aldose phase of fructose + aldose combinations suggested their possible involvement in governing the preferential utilization. The novel aspects of this study are enhanced organic acid mediated P solubilization in fructose + aldose combinations and a rare hierarchy of aldoses over fructose which is possibly regulated at the level of fructose transport and fructokinase. KEY POINTS: • Sugars when provided in different dual combinations, supported enhanced P solubilization from complex phosphate sources like TCP and RP in Rhizobium sp. RM. • Transcriptional status of genes in cells of RM when grown in different individual sugars and their combinations suggested that fructose might be a less preferred carbon source and hence was utilized after aldoses with the possible regulation by Hfq and HPrK. • First study to present a unique phenomenon of sequential utilization of aldoses (glucose, arabinose and xylose) over fructose in a concentration-independent manner in Rhizobium sp. RM. and to present the effect of dual combinations of sugars on organic acid mediated P solubilization trait of rhizobia.
Topics: Arabinose; Fructokinases; Fructose; Glucose; Organic Chemicals; Phosphates; Rhizobium; Xylose
PubMed: 35661910
DOI: 10.1007/s00253-022-11997-w -
Environmental Microbiology Feb 2023The Pseudomonas putida group in the Gammaproteobacteria has been intensively studied for bioremediation and plant growth promotion. Members of this group have recently...
The Pseudomonas putida group in the Gammaproteobacteria has been intensively studied for bioremediation and plant growth promotion. Members of this group have recently emerged as promising hosts to convert intermediates derived from plant biomass to biofuels and biochemicals. However, most strains of P. putida cannot metabolize pentose sugars derived from hemicellulose. Here, we describe three isolates that provide a broader view of the pentose sugar catabolism in the P. putida group. One of these isolates clusters with the well-characterized P. alloputida KT2440 (Strain BP6); the second isolate clustered with plant growth-promoting strain P. putida W619 (Strain M2), while the third isolate represents a new species in the group (Strain BP8). Each of these isolates possessed homologous genes for oxidative xylose catabolism (xylDXA) and a potential xylonate transporter. Strain M2 grew on arabinose and had genes for oxidative arabinose catabolism (araDXA). A CRISPR interference (CRISPRi) system was developed for strain M2 and identified conditionally essential genes for xylose growth. A glucose dehydrogenase was found to be responsible for initial oxidation of xylose and arabinose in strain M2. These isolates have illuminated inherent diversity in pentose catabolism in the P. putida group and may provide alternative hosts for biomass conversion.
Topics: Pentoses; Xylose; Arabinose; Pseudomonas putida; Oxidative Stress
PubMed: 36465038
DOI: 10.1111/1462-2920.16296 -
Spectrochimica Acta. Part A, Molecular... Nov 2019FTIR and NMR spectra were measured in parallel for specific two-components mixtures of various proteins with different sugar molecules, such as arabinose, glucose, and...
FTIR and NMR spectra were measured in parallel for specific two-components mixtures of various proteins with different sugar molecules, such as arabinose, glucose, and sucrose. In the FTIR spectra of arabinose with some of these proteins, the bands assigned to the vibrational modes of the CH and COH groups disappeared, and new ones, related to an arabinose-protein CN mode, appeared. Similar changes were observed in the FTIR spectra of lyophilized mixtures of arabinose with different amino acids. In additional FTIR spectra, measured for other protein-sugar mixtures, the bands correlated to the ring modes of arabinose, in the range 1150-1000 cm, disappeared, and two new very strong narrow bands became dominant, indicating ring opening or some kind of arabinose decomposition. Contrary to the prevailing opinion that complexes between sugars and proteins are formed mainly by hydrogen bonds, the IR and NMR spectra of the sugar-protein mixtures studied here suggest that significant chemical reactions also take place between the interacting sugar and the protein. Two types of sugar-protein chemical reactions can be distinguished on the basis of these IR spectra, leading to the formation of a new CN bond and to the decomposition of sugar skeletal bonds. The new IR bands suggest that the latter reaction results in the formation of new bonds, which are related to new polyether moieties. These results highlight the often ignored non-specific chemical reactions that take place between sugars and proteins, and demonstrate that the simultaneous application of FTIR and NMR spectroscopic analyses can detect and further characterize these types of sugar-protein interactions.
Topics: Arabinose; Cold Temperature; Glucose; Humans; Maillard Reaction; Nuclear Magnetic Resonance, Biomolecular; Proteins; Spectroscopy, Fourier Transform Infrared; Sucrose; Sugars; Xylose
PubMed: 31255896
DOI: 10.1016/j.saa.2019.02.085 -
Journal of the Science of Food and... Jul 2023The nixtamalization process improves the nutritional and technological properties of maize. This process generates nixtamalized maize bran as a by-product, which is a...
BACKGROUND
The nixtamalization process improves the nutritional and technological properties of maize. This process generates nixtamalized maize bran as a by-product, which is a source of arabinoxylans (AX). AX are polysaccharides constituted of a xylose backbone with mono- or di-arabinose substitutions, which can be ester-linked to ferulic acid (FA). The present study investigated the fine structural features and antioxidant capacity (AC) of nixtamalized maize bran arabinoxylans (MBAX) to comprehend the structure-radical scavenging capacity relationship in this polysaccharide deeply.
RESULTS
MBAX presented a molecular weight, intrinsic viscosity, and hydrodynamic radius of 674 kDa, 1.8 dL g , and 24.6 nm, respectively. The arabinose-to-xylose ratio (A/X) and FA content were 0.74 and 0.25 g kg polysaccharide, respectively. MBAX contained dimers (di-FA) and trimer (tri-FA) of FA (0.14 and 0.07 g kg polysaccharide, respectively). The main di-FA isomer was the 8-5' structure (80%). Fourier transform infrared spectroscopy confirmed MBAX molecular identity, and the second derivate of the spectral data revealed a band at 958 cm related to the presence of arabinose disubstitution. H-Nuclear magnetic resonance spectroscopy showed mono- and di-arabinose substitution in the xylan backbone with more monosubstituted residues. MBAX registered an AC of 25 and 20 μmol Trolox equivalents g polysaccharide despite a low FA content, using ABTS (2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid) and DPPH (1,1-diphenyl-2-picrylhydrazyl) methods, respectively.
CONCLUSION
AC in MBAX could be related to the high A/X ratio (mainly monosubstitution) and the high 8-5' di-FA proportion in this polysaccharide. © 2023 Society of Chemical Industry.
Topics: Xylans; Antioxidants; Zea mays; Xylose; Arabinose; Polysaccharides
PubMed: 36852427
DOI: 10.1002/jsfa.12531 -
Bioresource Technology May 2020Alpha-L-arabinofuranoside arabinofuranohydrolase (ARA), more commonly known as alpha-L-arabinofuranosidase (E.C. number 3.2.1.55), is a hydrolytic enzyme, catalyzing the... (Review)
Review
Alpha-L-arabinofuranoside arabinofuranohydrolase (ARA), more commonly known as alpha-L-arabinofuranosidase (E.C. number 3.2.1.55), is a hydrolytic enzyme, catalyzing the cleavage of alpha-L-arabinose by acting on the non-reducing ends of alpha-L-arabinofuranosides, alpha-L-arabinans containing (1,3)- and/or (1,5)-linked arabinoxylans and arabinogalactans. ARA functions as debranching enzyme removing arabinose substituents from arabinoxylan and arabinoxylooligomers, thereby, boosting the hydrolysis of arabinoxylan fraction of hemicellulose and improving bioconversion of lignocellulosic biomass. Previously, comprehensive information on this enzyme has not been reviewed thoroughly. Therefore, the main aim of this review is to highlight the important properties of this interesting enzyme, microorganisms used for its production, and enhanced production using genetic engineering approach. An account on synergism with other biomass hydrolyzing enzymes and various industrial applications of this enzyme has also been provided along with an outlook on further research and development.
Topics: Arabinose; Biomass; Glycoside Hydrolases; Hydrolysis; Substrate Specificity; Xylans
PubMed: 32089440
DOI: 10.1016/j.biortech.2020.123019 -
Food & Function Mar 2021l-Arabinose is a kind of plant-specific five-carbon aldose with benefits in type 2 diabetes mellitus. It has been shown to have good properties in improving glucose...
l-Arabinose is a kind of plant-specific five-carbon aldose with benefits in type 2 diabetes mellitus. It has been shown to have good properties in improving glucose homeostasis, but the underlying molecular mechanisms are still not clear. Hepatic gluconeogenesis is critical for regulating glucose homeostasis. Here, this study aimed to investigate whether l-arabinose could improve glucose metabolism via suppressing hepatic gluconeogenesis. High-fat-high-sucrose diet (HFHSD) or high-sucrose diet (HSD)-fed mice were supplemented with or without l-arabinose for 12 weeks. Fasting blood glucose levels were measured and glucose tolerance test and the histological analysis were performed after l-arabinose administration. AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), peroxisome proliferator activated receptor-γ coactivator-1α (PGC1α), Forkhead box O1 (FoxO1), phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) expression levels were determined by RT-PCR and western blotting. As expected, l-arabinose apparently decreased body weight and attenuated hyperglycemia and glucose intolerance caused by HFHSD or HSD. l-Arabinose also had beneficial effects on glycogen synthesis by inactivating GSK3β. The expression levels of gluconeogenic genes were all decreased by l-arabinose administration in vivo and in vitro. In addition, our work revealed that AMPK is required for the inhibitory effects of l-arabinose on hepatic gluconeogenesis. l-Arabinose significantly up-regulated the phosphorylated levels of AMPK and its downstream protein ACC. Furthermore, blocking AMPK signaling through an inhibitor (compound C) or siAMPK significantly attenuated the inhibition of hepatic gluconeogenesis and the promotion of glycogen synthesis with l-arabinose, indicating that the inhibitory effect of l-arabinose on hepatic gluconeogenesis was AMPK dependent. Our work revealed that l-arabinose is a promising natural product for the regulation of hyperglycemia through inhibition of hepatic gluconeogenesis by activating AMPK.
Topics: AMP-Activated Protein Kinases; Animals; Arabinose; Blood Glucose; Disease Models, Animal; Gluconeogenesis; Hyperglycemia; Male; Mice; Mice, Inbred C57BL
PubMed: 33502423
DOI: 10.1039/d0fo02163f -
Biochemical and Biophysical Research... Sep 2020L-Arabinose 1-dehydrogenase (AraDH) is responsible for the first step of the non-phosphorylative L-arabinose pathway from bacteria, and catalyzes the NAD(P)-dependent...
L-Arabinose 1-dehydrogenase (AraDH) is responsible for the first step of the non-phosphorylative L-arabinose pathway from bacteria, and catalyzes the NAD(P)-dependent oxidation of L-arabinose to L-arabinonolactone. This enzyme belongs to the so-called Gfo/Idh/MocA protein superfamily, but has a very poor phylogenetic relationship with other functional members. We previously reported the crystal structures of AraDH without a ligand and in complex with NADP. To clarify the underlying catalytic mechanisms in more detail, we herein elucidated the crystal structure in complex with L-arabinose and NADP. In addition to the previously reported five amino acid residues (Lys91, Glu147, His153, Asp169, and Asn173), His119, Trp152, and Trp231 interacted with L-arabinose, which were not found in substrate recognition by other Gfo/Idh/MocA members. Structure-based site-directed mutagenic analyses suggested that Asn173 plays an important role in catalysis, whereas Trp152, Trp231, and His119 contribute to substrate binding. The preference of NADP over NAD was significantly subjected by a pair of Ser37 and Arg38, whose manners were similar to other Gfo/Idh/MocA members.
Topics: Amino Acid Sequence; Arabinose; Azospirillum brasilense; Bacterial Proteins; Carbohydrate Dehydrogenases; Crystallography, X-Ray; Models, Molecular; NADP; Protein Conformation
PubMed: 32828286
DOI: 10.1016/j.bbrc.2020.07.071 -
Applied Microbiology and Biotechnology Mar 2016Bioprospecting is an effective way to find novel enzymes from strains with desirable phenotypes. Such bioprospecting has enabled organisms such as Saccharomyces...
Bioprospecting is an effective way to find novel enzymes from strains with desirable phenotypes. Such bioprospecting has enabled organisms such as Saccharomyces cerevisiae to utilize nonnative pentose sugars. Yet, the efficiency of this pentose catabolism (especially for the case of arabinose) remains suboptimal. Thus, further pathway optimization or identification of novel, optimal pathways is needed. Previously, we identified a novel set of xylan catabolic pathway enzymes from a superior pentose-utilizing strain of Ustilago bevomyces. These enzymes were used to successfully engineer a xylan-utilizing S. cerevisiae through a blended approach of bioprospecting and evolutionary engineering. Here, we expanded this approach to xylose and arabinose catabolic pathway engineering and demonstrated that bioprospected xylose and arabinose catabolic pathways from U. bevomyces offer alternative choices for enabling efficient pentose catabolism in S. cerevisiae. By introducing a novel set of xylose catabolic genes from U. bevomyces, growth rates were improved up to 85 % over a set of traditional Scheffersomyces stipitis pathway genes. In addition, we suggested an alternative arabinose catabolic pathway which, after directed evolution and pathway engineering, enabled S. cerevisiae to grow on arabinose as a sole carbon source in minimal medium with growth rates upwards of 0.05 h(-1). This pathway represents the most efficient growth of yeast on pure arabinose minimal medium. These pathways provide great starting points for further strain development and demonstrate the utility of bioprospecting from U. bevomyces.
Topics: Arabinose; Bioprospecting; Carbohydrates; Culture Media; Cytosol; Enzymes; Metabolic Engineering; Metabolic Networks and Pathways; Recombinant Proteins; Saccharomyces cerevisiae; Ustilago; Xylans; Xylose
PubMed: 26671616
DOI: 10.1007/s00253-015-7211-z -
Applied and Environmental Microbiology Nov 2021PcAxy43B is a modular protein comprising a catalytic domain of glycoside hydrolase family 43 (GH43), a family 6 carbohydrate-binding module (CBM6), and a family 36...
A Novel Multifunctional Arabinofuranosidase/Endoxylanase/β-Xylosidase GH43 Enzyme from Paenibacillus curdlanolyticus B-6 and Its Synergistic Action To Produce Arabinose and Xylose from Cereal Arabinoxylan.
PcAxy43B is a modular protein comprising a catalytic domain of glycoside hydrolase family 43 (GH43), a family 6 carbohydrate-binding module (CBM6), and a family 36 carbohydrate-binding module (CBM36) and found to be a novel multifunctional xylanolytic enzyme from Paenibacillus curdlanolyticus B-6. This enzyme exhibited α-l-arabinofuranosidase, endoxylanase, and β-d-xylosidase activities. The α-l-arabinofuranosidase activity of PcAxy43B revealed a new property of GH43, via the release of both long-chain cereal arabinoxylan and short-chain arabinoxylooligosaccharide (AXOS), as well as release from both the C(O) and C(O) positions of AXOS, which is different from what has been seen for other arabinofuranosidases. PcAxy43B liberated a series of xylooligosaccharides (XOSs) from birchwood xylan and xylohexaose, indicating that PcAxy43B exhibited endoxylanase activity. PcAxy43B produced xylose from xylobiose and reacted with -nitrophenyl-β-d-xylopyranoside as a result of β-xylosidase activity. PcAxy43B effectively released arabinose together with XOSs and xylose from the highly arabinosyl-substituted rye arabinoxylan. Moreover, PcAxy43B showed significant synergistic action with the trifunctional endoxylanase/β-xylosidase/α-l-arabinofuranosidase PcAxy43A and the endoxylanase Xyn10C from strain B-6, in which almost all products produced from rye arabinoxylan by these combined enzymes were arabinose and xylose. In addition, the presence of CBM36 was found to be necessary for the endoxylanase property of PcAxy43B. PcAxy43B is capable of hydrolyzing untreated cereal biomass, corn hull, and rice straw into XOSs and xylose. Hence, PcAxy43B, a significant accessory multifunctional xylanolytic enzyme, is a potential candidate for application in the saccharification of cereal biomass. Enzymatic saccharification of cereal biomass is a strategy for the production of fermented sugars from low-price raw materials. In the present study, PcAxy43B from B-6 was found to be a novel multifunctional α-l-arabinofuranosidase/endoxylanase/β-d-xylosidase enzyme of glycoside hydrolase family 43. It is effective in releasing arabinose, xylose, and XOSs from the highly arabinosyl-substituted rye arabinoxylan, which is usually resistant to hydrolysis by xylanolytic enzymes. Moreover, almost all products produced from rye arabinoxylan by the combination of PcAxy43B with the trifunctional xylanolytic enzyme PcAxy43A and the endoxylanase Xyn10C from strain B-6 were arabinose and xylose, which can be used to produce several value-added products. In addition, PcAxy43B is capable of hydrolyzing untreated cereal biomass into XOSs and xylose. Thus, PcAxy43B is an important multifunctional xylanolytic enzyme with high potential in biotechnology.
Topics: Arabinose; Bacterial Proteins; Edible Grain; Endo-1,4-beta Xylanases; Glycoside Hydrolases; Multifunctional Enzymes; Paenibacillus; Xylans; Xylose; Xylosidases
PubMed: 34613758
DOI: 10.1128/AEM.01730-21