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Journal of Food Biochemistry Dec 2021L-arabinose is a good and healthy food additive. This study was conducted to investigate the effect of L-arabinose in a mouse model of type 2 diabetes mellitus (T2DM)...
L-arabinose is a good and healthy food additive. This study was conducted to investigate the effect of L-arabinose in a mouse model of type 2 diabetes mellitus (T2DM) induced by exposure to a high-fat diet (HFD) and streptozotocin (STZ). The model mice received L-arabinose at 20 and 60 mg (kg body weight [bw]) d , metformin at 300 mg (kg bw) d (positive control) or sterile water (control) via oral gavage. Compared with the model group, mice treated with L-arabinose exhibited attenuated symptoms of diabetes mellitus, including a slower rate of body weight loss, increased homeostasis model assessment of β-cell function index levels, decreased blood glucose, alleviation of steatosis, and repair of pancreatic islet cells. L-arabinose also exerted an anti-inflammatory effect and partially mitigated dyslipidemia. A 16S-rRNA sequence analysis of the gut microbiota revealed that at the phylum level, treatment with L-arabinose significantly reduced the ratio of Firmicutes to Bacteroidetes due to a decreased relative abundance of Firmicutes; at the genus level, it reversed the increase in the relative abundance of Allobaculum and the decrease abundance of Oscillospira caused by exposure to an HFD and STZ. And the model mice received L-arabinose at 20 mg (kg bw) d had a better effect on improving T2DM than the high-dose group supplemented L-arabinose at 60 mg (kg bw) d . These results strongly suggest L-arabinose as an excellent candidate supplement to prevent or treat T2DM. PRACTICAL APPLICATIONS: L-arabinose, xylitol and sucralose are well-known substitutes for sucrose. L-arabinose has been reported to have beneficial effects on hyperglycemia, glycemic index, and fat accumulation. In this study, we found that low-dose (20 mg (kg bw) d ) supplementation of L-arabinose significantly improved glucose intolerance and gut microbiota incoordination in T2DM caused by HFD and STZ.
Topics: Animals; Arabinose; Diabetes Mellitus, Type 2; Diet, High-Fat; Gastrointestinal Microbiome; Hypoglycemic Agents; Mice; Mice, Inbred C57BL; Streptozocin
PubMed: 34778991
DOI: 10.1111/jfbc.13991 -
Biotechnology Reports (Amsterdam,... Jun 2021d-xylose reductase is a member of the aldo-keto reductase family, and is involved in d-xylose and l-arabinose conversion through the Pentose Catabolic Pathway (PCP) in...
d-xylose reductase is a member of the aldo-keto reductase family, and is involved in d-xylose and l-arabinose conversion through the Pentose Catabolic Pathway (PCP) in fungi. In this study, we biochemically characterized a newly identified second d-xylose reductase (XyrB) from . This NADPH-dependent reductase is able to efficiently convert d-xylose and l-arabinose, and it has the highest affinity for these sugars of all currently known fungal pentose reductases. A combination of biochemical data, transcriptomics and phylogenetic analysis further illustrated the role of XyrB in the PCP. Enzymes: D-xylose reductase (EC 1.1.1.307), L-arabinose reductase (EC 1.1.1.21).
PubMed: 33842213
DOI: 10.1016/j.btre.2021.e00610 -
EcoSal Plus Dec 2022Very few labs have had the good fortune to have been able to focus for more than 50 years on a relatively narrow research topic and to be in a field in which both... (Review)
Review
Very few labs have had the good fortune to have been able to focus for more than 50 years on a relatively narrow research topic and to be in a field in which both basic knowledge and the research technology and methods have progressed as rapidly as they have in molecular biology. My research group, first at Brandeis University and then at Johns Hopkins University, has had this opportunity. In this review, therefore, I will describe largely the work from my laboratory that has spanned this period and which was carried out by 40 plus graduate students, several postdoctoral associates, my technician, and me. In addition to presenting the scientific findings or results, I will place many of the topics in scientific context and, because we needed to develop a good many of the experimental methods behind our findings, I will also describe some of these methods and their importance. Also included will be occasional comments on how the research community or my research group functioned. Because a wide variety of approaches were used throughout our work, no ideal organization of this review is apparent. Therefore, I have chosen to use a hybrid structure in which there are six sections. Within each of the sections, experiments and findings will be described roughly in chronological order. Frequent cross references between parts and sections will be made because some findings and experimental approaches could logically have been described in more than one place.
Topics: Humans; Arabinose; Learning; Technology; Operon
PubMed: 36519894
DOI: 10.1128/ecosalplus.ESP-0012-2021 -
Cell Host & Microbe Mar 2023The molecular understanding of host-pathogen interactions in the gastrointestinal (GI) tract of superspreader hosts is incomplete. In a mouse model of chronic,...
The molecular understanding of host-pathogen interactions in the gastrointestinal (GI) tract of superspreader hosts is incomplete. In a mouse model of chronic, asymptomatic Salmonella enterica serovar Typhimurium (S. Tm) infection, we performed untargeted metabolomics on the feces of mice and found that superspreader hosts possess distinct metabolic signatures compared with non-superspreaders, including differential levels of L-arabinose. RNA-seq on S. Tm from superspreader fecal samples showed increased expression of the L-arabinose catabolism pathway in vivo. By combining bacterial genetics and diet manipulation, we demonstrate that diet-derived L-arabinose provides S. Tm a competitive advantage in the GI tract, and expansion of S. Tm in the GI tract requires an alpha-N-arabinofuranosidase that liberates L-arabinose from dietary polysaccharides. Ultimately, our work shows that pathogen-liberated L-arabinose from the diet provides a competitive advantage to S. Tm in vivo. These findings propose L-arabinose as a critical driver of S. Tm expansion in the GI tracts of superspreader hosts.
Topics: Salmonella typhimurium; Arabinose; Salmonella enterica; Polysaccharides; Serogroup
PubMed: 36812913
DOI: 10.1016/j.chom.2023.01.017 -
Biotechnology For Biofuels and... May 2022Owing to increasing concerns about climate change and the depletion of fossil fuels, the development of efficient microbial processes for biochemical production from...
BACKGROUND
Owing to increasing concerns about climate change and the depletion of fossil fuels, the development of efficient microbial processes for biochemical production from lignocellulosic biomass has been a key issue. Because process efficiency is greatly affected by the inherent metabolic activities of host microorganisms, it is essential to utilize a microorganism that can rapidly convert biomass-derived sugars. Here, we report a novel Vibrio-based microbial platform that can rapidly and simultaneously consume three major lignocellulosic sugars (i.e., glucose, xylose, and arabinose) faster than any previously reported microorganisms.
RESULTS
The xylose isomerase pathway was constructed in Vibrio sp. dhg, which naturally displays high metabolic activities on glucose and arabinose but lacks xylose catabolism. Subsequent adaptive laboratory evolution significantly improved xylose catabolism of initial strain and led to unprecedently high growth and sugar uptake rate (0.67 h and 2.15 g g h, respectively). Furthermore, we achieved co-consumption of the three sugars by deletion of PtsG and introduction of GalP. We validated its superior performance and applicability by demonstrating efficient lactate production with high productivity (1.15 g/L/h) and titer (83 g/L).
CONCLUSIONS
In this study, we developed a Vibrio-based microbial platform with rapid and simultaneous utilization of the three major sugars from lignocellulosic biomass by applying an integrated approach of rational and evolutionary engineering. We believe that the developed strain can be broadly utilized to accelerate the production of diverse biochemicals from lignocellulosic biomass.
PubMed: 35614459
DOI: 10.1186/s13068-022-02157-3 -
Colloids and Surfaces. B, Biointerfaces Sep 2022The biocatalysts are broadly explored in the biological transformation processes. The enzyme cascade catalysis involves various catalytic activities in a sequential...
The biocatalysts are broadly explored in the biological transformation processes. The enzyme cascade catalysis involves various catalytic activities in a sequential process to produce the desired product including the formation of reaction intermediates. Enzyme immobilization is a method in which enzymes are confined within a support or matrix either physically or chemically to enhance their relative stability and catalytic activity in the enzyme cascade catalysis. In view of this, L-arabinose isomerase (L-AI) and L-ribose isomerase (L-RI) were immobilized on zeolite based metal framework as a micro-composite construct (DEMC@L-AI+L-RI) using linker, and metal ions. Such immobilization could be of great significance and provide several advantages like mesoporous surface for enzyme adsorption, desirable functionality in the production of products in enzyme cascade reaction, high storage stability and enhanced recyclability. The developed DEMC@L-AI+L-RI was characterized using SEM, FTIR, CLSM and TGA. The immobilization yield was 32% and loading of enzyme was 22% on the surface of micro-composite. The DEMC@L-AI+L-RI showed relatively stable catalytic activity at pH 5-6 and temperature 40 °C. The catalytic efficiency (k/K) of both the enzymes was increased by 1.5-fold after immobilization. With the immobilized biocatalyst, bioconversion of L-arabinose to L-ribose was 22.6% and D-galactose to D-talose was 15.2%. The reusability of developed biocatalyst for more than six cycles was observed for more than 50% yield of the sugars. The conversion of biomass sugars from beetroot and onion waste residues was 20% and 14% to produce ribose and talose, respectively.
Topics: Aldose-Ketose Isomerases; Hexoses; Hydrogen-Ion Concentration; Lactones; Metals; Ribose
PubMed: 35728372
DOI: 10.1016/j.colsurfb.2022.112637 -
Polymers Jul 2022This study focuses on the acetylation modification of polysaccharides from leaves (RDPs) with a high degree of substitution (DS) and then discusses their...
This study focuses on the acetylation modification of polysaccharides from leaves (RDPs) with a high degree of substitution (DS) and then discusses their characterization and biological activity. The optimum acetylation conditions of RDPs were optimized by response surface methodology, which were reaction time 3 h, reaction temperature 50 °C, and the liquid-solid ratio 16 mL/g. Under the optima schemes, two eco-friendly acetylated polysaccharides from leaves (AcRDP-1 with DS of 0.439 ± 0.025 and AcRDP-2 with DS of 0.445 ± 0.022) were prepared. The results of structural characterization showed that the AcRDP-1 (9.3525 × 10 kDa) and AcRDP-2 (4.7016 × 10 kDa) were composed of mannose, glucose, galactose, and arabinose with molar ratios of 1.00:5.01:1.17:0.15 and 1.00:4.47:2.39:0.88, respectively. Compared with unmodified polysaccharides, the arabinose content and molecular weight of the two acetylated polysaccharides decreased, and their triple helix conformation disappeared, and further improved their anticomplementary activity. The two acetylated polysaccharides showed stronger a complement inhibition effect than the positive drug by blocking C2, C3, C4, C5, C9, and factor B targets in the classical and alternative pathways. This research indicated that acetylation modification could effectively enhance the anticomplementary activity of RDPs, which is beneficial for the development and utilization of leaves.
PubMed: 35956644
DOI: 10.3390/polym14153130 -
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 -
Methods in Enzymology 2021Since its invention, recombinant protein expression has greatly facilitated our understanding of various cellular processes in different biological systems because...
Since its invention, recombinant protein expression has greatly facilitated our understanding of various cellular processes in different biological systems because theoretically this technique renders any gene to be expressed in a mesophilic host like Escherichia coli, thus allowing functional characterizations of proteins of interest. However, such a practice has only yielded a limited success for proteins encoded in thermophilic archaea since thermophilic proteins are often present in an insoluble form when expressed in E. coli. As a result, it is advantageous to express recombinant proteins of thermophilic archaea in a homologous host, allowing a native form of recombinant protein to be purified and characterized. Here we present a detailed protocol for the homologous expression and purification of proteins in the thermophilic archaeon, Sulfolobus islandicus Rey15A.
Topics: Archaea; Escherichia coli; Protein Processing, Post-Translational; Recombinant Proteins; Sulfolobus
PubMed: 34752289
DOI: 10.1016/bs.mie.2021.05.006