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1.
Microb Cell Fact ; 21(1): 154, 2022 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-35933385

RESUMO

BACKGROUND: Sugar hydrolysates from lignocellulosic biomass are majorly composed of glucose and xylose that can be fermented to biofuels. Bacteria, despite having the natural ability to consume xylose are unable to consume it in presence of glucose due to a carbon catabolite repression (CCR) mechanism. This leads to overall reduced productivity as well as incomplete xylose utilization due to ethanol build-up from glucose utilization. In our effort to develop a strain for simultaneous fermentation of glucose and xylose into ethanol, we deleted ptsG in ethanologenic E. coli SSK42 to make it deficient in CCR and performed adaptive laboratory evolution to achieve accelerated growth rate, sugar consumption and ethanol production. Finally, we performed proteomics study to identify changes that might have been responsible for the observed improved phenotype of the evolved strain. RESULTS: The parental strain of SSK42, i.e., wild-type E. coli B, did not co-utilize glucose and xylose as expected. After deleting the ptsG gene encoding the EIIBCGlc subunit of PTS system, glucose consumption is severely affected in wild-type E. coli B. However, the ethanologenic, SSK42 strain, which was evolved in our earlier study on both glucose and xylose, didn't show such a drastic effect of EIIBCGlc deletion, instead consumed glucose first, followed by xylose without delay for switching from one sugar to another. To improve growth on xylose and co-utilization capabilities, the ptsG deleted SSK42 was evolved on xylose. The strain evolved for 78 generations, strain SCD78, displayed significant co-utilization of glucose and xylose sugars. At the bioreactor level, the strain SCD78 produced 3-times the ethanol titer of the parent strain with significant glucose-xylose co-utilization. The rate of glucose and xylose consumption also increased 3.4-fold and 3-fold, respectively. Proteome data indicates significant upregulation of TCA cycle proteins, respiration-related proteins, and some transporters, which may have a role in increasing the total sugar consumption and co-utilization of sugars. CONCLUSION: Through adaptive evolution, we have obtained a strain that has a significant glucose-xylose co-utilization phenotype with 3-fold higher total sugar consumption rate and ethanol production rate compared to the unevolved strain. This study also points out that adaptation on xylose is enough to impart glucose-xylose co-utilization property in CCR compromised ethanologenic strain SSK42.


Assuntos
Repressão Catabólica , Xilose , Açúcares da Dieta/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Etanol/metabolismo , Fermentação , Glucose/metabolismo , Açúcares/metabolismo , Xilose/metabolismo
2.
Enzyme Microb Technol ; 160: 110086, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35785629

RESUMO

In order to better understand the function of a putative GH8 xylanase gene in the xylan-degrading bacterium Paenibacillus physcomitrellae XB, a novel reducing-end xylose-releasing exo-oligoxylanase (Rex) PphRex8A of GH8 family was characterized. Phylogenetic analysis showed that it was clustered tightly with other published GH8 Rexs and exhibited the highest amino acid sequence identity (77.4 %) with the Rex of PbRex8 from P. barengoltzii G22. The three-dimensional (3D) structure of PphRex8A was also built based on the template of PbRex8 (5YXT) and three conserved catalytic active sites (Glu71, Asp263, and Asp129) were predicted and further confirmed by the enzymatic inactivity of their mutants (E71A, D129A, and D263A). The hydrolysis assay of PphRex8A showed that it could hydrolyze xylo-oligosaccharides (XOSs) with a degree of polymerization (DP) ≥ 3, such as xylotriose (X3) through xylohexaose (X6), and some natural XOSs, such as corncob xylan (CCX) and oat spelt xylan (OSX) to release xylose. However, it could not hydrolyze p-nitrophenyl-ß-D-xylopyranoside (pNPX), which suggested that it mainly released xylose from the reducing-end of XOSs and belonged to a Rex. In addition, PphRex8A also could deconstruct xylans with high DP, such as wheat arabinoxylan (AX) and beech wood xylan (BWX) to produce XOSs with DP3-6. Moreover, PphRex8A had synergistic effects with other xylanolytic enzymes of P. physcomitrellae XB, such as with PphXyn10 or PphXyn11 at a ratio of 1:3, or with PphXyl43B as a ratio of 3:1, significantly increasing the amounts of reducing sugars toward different xylan substrates. Thus, PphRex8A could be an exo-xylanase toward XOSs and could improve the deconstruction capability of high DP xylans, thereby complementing other xylanolytic enzymes to contribute to xylan degradation and improve the efficiency of converting hemicellulose biomass into energy by P. physcomitrellae XB.


Assuntos
Xilanos , Xilose , Endo-1,4-beta-Xilanases/metabolismo , Hidrólise , Oligossacarídeos/metabolismo , Paenibacillus , Filogenia , Especificidade por Substrato , Xilanos/metabolismo , Xilose/metabolismo
3.
Enzyme Microb Technol ; 160: 110094, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35810624

RESUMO

Lignocellulose hydrolysates are rich in fermentable sugars such as xylose, cellobiose and glucose, with high potential in the biotechnology industry to obtain bioproducts of higher economic value. Thus, it is important to search for and study new yeast strains that co-consume these sugars to achieve better yields and productivity in the processes. The yeast Clavispora lusitaniae CDBB-L-2031, a native strain isolated from mezcal must, was studied under various culture conditions to potentially produce ethanol and xylitol due to its ability to assimilate xylose, cellobiose and glucose. This yeast produced ethanol under microaerobic conditions with yields of 0.451 gethanol/gglucose and 0.344 gethanol/gcellobiose, when grown on 1% glucose or cellobiose, respectively. In mixtures (0.5% each) of glucose:xylose and glucose:xylose:cellobiose the yields were 0.367 gethanol/gGX and 0. 380 gethanol/gGXC, respectively. Likewise, in identical conditions, C. lusitaniae produced xylitol from xylose with a yield of 0.421 gxylitol/gxylose. In 5% glucose or xylose, this yeast had better ethanol and xylitol titers and yields, respectively. However, glucose negatively affected xylitol production in the mixture of both sugars (3% each), producing only ethanol. Xylose reductase (XR) and xylitol dehydrogenase (XDH) activities were evaluated in cultures growing on xylose or glucose, obtaining the highest values in cultures on xylose at 8 h (25.9 and 6.22 mU/mg, respectively). While in glucose cultures, XR and XDH activities were detected once this substrate was consumed (4.06 and 3.32 mU/mg, respectively). Finally, the XYL1 and XYL2 genes encoding xylose reductase and xylitol dehydrogenase, respectively, were up-regulated by xylose, whereas glucose down-regulated their expression.


Assuntos
Xilitol , Xilose , Aldeído Redutase/genética , Aldeído Redutase/metabolismo , Celobiose/metabolismo , D-Xilulose Redutase/genética , D-Xilulose Redutase/metabolismo , Etanol/metabolismo , Fermentação , Glucose/metabolismo , Saccharomyces cerevisiae/genética , Saccharomycetales , Xilitol/metabolismo , Xilose/metabolismo
4.
BMC Microbiol ; 22(1): 173, 2022 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-35799117

RESUMO

BACKGROUND: Sustainable production of oil for food, feed, fuels and other lipid-based chemicals is essential to meet the demand of the increasing human population. Consequently, novel and sustainable resources such as lignocellulosic hydrolysates and processes involving these must be explored. In this paper we screened for naturally-occurring xylose utilizing oleaginous yeasts as cell factories for lipid production, since pentose sugar catabolism plays a major role in efficient utilization of lignocellulosic feedstocks. Glycerol utilization, which is also beneficial in yeast-based oil production as glycerol is a common by-product of biodiesel production, was investigated as well. Natural yeast isolates were studied for lipid accumulation on a variety of substrates, and the highest lipid accumulating strains were further investigated in shake flask cultivations and fermenter studies on xylose and hydrolysate. RESULTS: By collecting leaves from exotic plants in greenhouses and selective cultivation on xylose, a high frequency of oleaginous yeasts was obtained (> 40%). Different cultivation conditions lead to differences in fatty acid contents and compositions, resulting in a set of strains that can be used to select candidate production strains for different purposes. In this study, the most prominent strains were identified as Pseudozyma hubeiensis BOT-O and Rhodosporidium toruloides BOT-A2. The fatty acid levels per cell dry weight after cultivation in a nitrogen limited medium with either glucose, xylose or glycerol as carbon source, respectively, were 46.8, 43.2 and 38.9% for P. hubeiensis BOT-O, and 40.4, 27.3 and 42.1% for BOT-A2. Furthermore, BOT-A2 accumulated 45.1% fatty acids per cell dry weight in a natural plant hydrolysate, and P. hubeiensis BOT-O showed simultaneous glucose and xylose consumption with similar growth rates on both carbon sources. The fatty acid analysis demonstrated both long chain and poly-unsaturated fatty acids, depending on strain and medium. CONCLUSIONS: We found various natural yeast isolates with high lipid production capabilities and the ability to grow not only on glucose, but also xylose, glycerol and natural plant hydrolysate. R. toruloides BOT-A2 and P. hubeiensis BOT-O specifically showed great potential as production strains with high levels of storage lipids and comparable growth to that on glucose on various other substrates, especially compared to currently used lipid production strains. In BOT-O, glucose repression was not detected, making it particularly desirable for utilization of plant waste hydrolysates. Furthermore, the isolated strains were shown to produce oils with fatty acid profiles similar to that of various plant oils, making them interesting for future applications in fuel, food or feed production.


Assuntos
Glicerol , Xilose , Carbono/metabolismo , Ácidos Graxos/metabolismo , Glucose/metabolismo , Humanos , Lipídeos/análise , Xilose/metabolismo , Leveduras/metabolismo
5.
J Agric Food Chem ; 70(27): 8409-8416, 2022 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-35771137

RESUMO

The Amadori rearrangement product derived from xylose-glycylglycine (XGG-ARP) is reactive to be attacked by another glycylglycine to generate a xylose-glycylglycine cross-linking product (XGG-CP) as a secondary product of the ARP. In this research, the role of additional glycylglycine in the XGG-ARP degradation was studied, and the dependence of glycylglycine on temperature was further clarified. The yields of XGG-CP and its degradation products were significantly affected by the molar ratio of glycylglycine to XGG-ARP. At the similar total concentration of reactant XGG-ARP and glycylglycine, the yields of XGG-CP, 3-deoxyxylosone, and furfural were dramatically decreased as the molar ratio of glycylglycine to XGG-ARP was increased. However, when the reaction temperature was increased to 120 °C, the increased additional glycylglycine percentage showed an obvious catalytic effect on the XGG-ARP degradation to deoxyosone and thus improved the furfural yield as well. The results revealed that an increased glycylglycine dosage level enhanced both the conversion of XGG-ARP to XGG-CP and the conversion of XGG-CP to 3-deoxyosone. The high-temperature-induced unequal acceleration for XGG-CP formation and degradation at a high glycylglycine dosage further led to a catalytic effect on the ARP degradation to deoxyosone. The concentration of 3-deoxyosone was increased by 37.5% when the molar ratio of glycylglycine to XGG-ARP increased from 1:2 to 2:1 at a temperature of 120 °C.


Assuntos
Glicilglicina , Reação de Maillard , Catálise , Furaldeído , Temperatura , Xilose
6.
BMC Microbiol ; 22(1): 164, 2022 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-35751041

RESUMO

BACKGROUND: 2,3-Butanediol (2,3-BD), a valuable compound used for chemicals, cosmetics, pesticides and pharmaceuticals, has been produced by various microbes. However, no high-temperature fermentation of the compound at high productivity has been reported. METHODS: Thermotolerant xylose-utilizing microbes were isolated from 6 different districts in Laos and screened for a low accumulation of xylitol in a xylose medium at 37 ˚C. One isolate was found to produce 2,3-BD and identified by 16S rDNA sequencing. The 2,3-BD fermentation capacity was investigated at different temperatures using xylose and glucose as carbon sources, and the fermentation parameters were determined by a high-performance liquid chromatography system. RESULTS: By screening for a low accumulation of xylitol in a xylose medium, one isolate that accumulated almost no xylitol was obtained. Further analyses revealed that the isolate is Cronobacter sakazakii and that it has the ability to produce 2,3-BD at high temperatures. When xylose and glucose were used, this strain, named C. sakazakii OX-25, accumulated 2,3-BD in a short period before the complete consumption of these sugars and then appeared to convert 2,3-BD to acetoin. The optimum temperature of the 2,3-BD fermentation was 42 ˚C to 45 ˚C, and the maximum yield of 2,3-BD was 0.3 g/g at 12 h in 20 g/l xylose medium and 0.4 g/g at 6 h in 20 g/l glucose medium at 42 ˚C. The 2,3-BD productivity of the strain was higher than the 2,3-BD productivities of other non-genetically engineered microorganisms reported previously, and the highest productivity was 0.6 g/l·h and 1.2 g/l·h for xylose and glucose, respectively. CONCLUSIONS: Among thermotolerant microbes isolated in Laos, we discovered a strain, C. sakazakii OX-25, that can convert xylose and glucose to 2,3-BD with high efficiency and high productivity at high temperatures, suggesting that C. sakazakii OX-25 has the potential for industrial application to produce 2,3-BD as an important platform chemical.


Assuntos
Cronobacter sakazakii , Xilose , Butileno Glicóis , Fermentação , Glucose/química , Xilitol
7.
Pharm Biol ; 60(1): 1237-1254, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35763552

RESUMO

CONTEXT: Hedysari Radix Praeparata Cum Melle (HRPCM) and Astragali Radix Praeparata Cum Melle (ARPCM) are used interchangeably in clinics to treat spleen-qi deficiency (SQD) symptom mainly including gastrointestinal dysfunction and decreased immunity, which has unknown differences in efficacy. OBJECTIVE: To investigate the differences between HRPCM and ARPCM on intervening gastrointestinal- and immune-function with SQD syndrome. MATERIALS AND METHODS: After the SQD model was established, the Sprague-Dawley (SD) rats were randomly divided into nine groups (n = 10): normal; model; Bu-Zhong-Yi-Qi Pills; 18.9, 12.6 and 6.3 g/kg dose groups of HRPCM and ARPCM. Gastrointestinal function including d-xylose, gastrin, amylase vasoactive intestinal peptide, motilin, pepsin, H+/K+-ATPase, Na+/K+-ATPase, sodium-glucose cotransporter 1 (SGLT1), glucose transporter 2 (GLUT2) and immune function including spleen and thymus index, blood routine, interleukin (IL)-2, IL-6, interferon-γ (IFN-γ), tumour necrosis factor-α (TNF-α), immunoglobulin (Ig) M, IgA, IgG and delayed-type hypersensitivity (DTH) were detected. Finally, the efficacy differences were analysed comprehensively by the fuzzy matter-element method. RESULTS: In regulating immune, the doses differences in efficacy between HRPCM and ARPCM showed in the high-dose (18.9 g/kg), but there were no differences in the middle- and low- dose (12.6 and 6.37 g/kg); the efficacy differences were primarily reflected in levels of IL-6, IFN-γ, TNF-α and IgM in serum, and the mRNA expression of IL-6 and IFN-γ in the spleen. In regulating gastrointestinal, the efficacy differences were primarily reflected in the levels of D-xylose, MTL, and GAS in serum, and the mRNA and protein expression of SGLT1 and GLUT2 in jejunum and ileum. DISCUSSION AND CONCLUSIONS: HRPCM is more effective than ARPCM on regulating gastrointestinal function and immune function with SQD syndrome. Therefore, we propose that HRPCM should be mainly used to treat SQD syndrome in the future.


Assuntos
Astrágalo (Planta) , Medicamentos de Ervas Chinesas , Adenosina Trifosfatases , Animais , Medicamentos de Ervas Chinesas/farmacologia , Interleucina-6 , RNA Mensageiro , Ratos , Ratos Sprague-Dawley , Baço , Fator de Necrose Tumoral alfa/farmacologia , Xilose/farmacologia
8.
PLoS One ; 17(6): e0269972, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35763520

RESUMO

Proteoglycan glycosaminoglycan (GAG) chains are attached to a serine residue in the protein through a linkage series of sugars, the first of which is xylose. Xylosides are chemicals which compete with the xylose at the enzyme xylosyl transferase to prevent the attachment of GAG chains to proteins. These compounds have been employed at concentrations in the millimolar range as tools to study the role of GAG chains in proteoglycan function. In the course of our studies with xylosides, we conducted a dose-response curve for xyloside actions on neural cells. To our surprise, we found that concentrations of xylosides in the nanomolar to micromolar range had major effects on cell morphology of hippocampal neurons as well as of Neuro2a cells, affecting both actin and tubulin cytoskeletal dynamics. Such effects/morphological changes were not observed with higher xyloside concentrations. We found a dose-dependent alteration of GAG secretion by Neuro2a cells; however, concentrations of xylosides which were effective in altering neuronal morphology did not cause a large change in the rate of GAG chain secretion. In contrast, both low and high concentrations of xylosides altered HS and CS composition. RNAseq of treated cells demonstrated alterations in gene expression only after treatment with millimolar concentration of xylosides that had no effect on cell morphology. These observations support a novel action of xylosides on neuronal cells.


Assuntos
Glicosídeos , Xilose , Glicosaminoglicanos/metabolismo , Glicosídeos/química , Proteoglicanas/metabolismo , Xilose/farmacologia
9.
Plant Foods Hum Nutr ; 77(2): 299-306, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35661961

RESUMO

Fruit processing by-products may be re-utilized as prebiotic ingredients to minimize the environmental impact of solid wastes generated from food industries. This study investigated the effects of enzymatic-induced hydrolysis on two types of guava purée by-products, particularly the prebiotic activity after its inclusion in yogurt-making. Commercial cellulase and xylanase were applied together or separately on refiner (the seed-rich fraction), and decanter (the pulp-rich fraction); labelled as 150 XY (xylanase); 150 CE (cellulase), 150 CX (combined cellulase-xylanase), and CT (control, untreated). The hydrolysis extents followed the order of 150 XY < 150 CE < 150CX. The ethanolic extracts (EEC) of the treated samples were analyzed on selected sugar content and the prebiotic activity score. Rhamnose and xylose were the main sugar constituents in both refiner and decanter. A two to four-fold increments of prebiotic activity score were observed on EEC of combined cellulase and xylanase treated decanter and refiner. Incorporating the combined enzymatically treated whole guava by-products into UHT fresh milk containing a yogurt starter culture significantly increased the log CFU/mL up to 77.6%, enhanced hardness, stickiness, and adhesiveness ranging from 22.2 to 86.4%, and decreased pH values. Combined cellulase-xylanase treatment can convert guava purée by-products into potential prebiotic sources for food applications.


Assuntos
Celulase , Psidium , Carboidratos , Celulase/química , Endo-1,4-beta-Xilanases/química , Hidrólise , Prebióticos , Psidium/química , Xilose , Iogurte
10.
World J Microbiol Biotechnol ; 38(7): 127, 2022 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-35668329

RESUMO

Interest in the production of renewable chemicals from biomass has increased in the past years. Among these chemicals, carboxylic acids represent a significant part of the most desirable bio-based products. Xylonic acid is a five-carbon sugar-acid obtained from xylose oxidation that can be used in several industrial applications, including food, pharmaceutical, and construction industries. So far, the production of xylonic acid has not yet been available at an industrial scale; however, several microbial bio-based production processes are under development. This review summarizes the recent advances in pathway characterization, genetic engineering, and fermentative strategies to improve xylonic acid production by microorganisms from xylose or lignocellulosic hydrolysates. In addition, the strengths of the available microbial strains and processes and the major requirements for achieving biotechnological production of xylonic acid at a commercial scale are discussed. Efficient native and engineered microbial strains have been reported. Xylonic acid titers as high as 586 and 171 g L-1 were obtained from bacterial and yeast strains, respectively, in a laboratory medium. Furthermore, relevant academic and industrial players associated with xylonic acid production will be presented.


Assuntos
Biotecnologia , Xilose , Biomassa , Fermentação , Engenharia Metabólica , Xilose/análogos & derivados , Xilose/metabolismo
11.
Appl Microbiol Biotechnol ; 106(11): 4251-4268, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35661910

RESUMO

Rhizobium sp. RM solubilized tri-calcium phosphate (TCP: 324-463 µg ml-1) and rock phosphate (RP: 36-46.58 µg ml-1) 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-1) and RP (75 µg ml-1) 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.


Assuntos
Rhizobium , Arabinose/metabolismo , Frutoquinases/metabolismo , Frutose/metabolismo , Glucose/metabolismo , Compostos Orgânicos/metabolismo , Fosfatos/metabolismo , Rhizobium/genética , Xilose/metabolismo
12.
Int J Biol Macromol ; 213: 902-914, 2022 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-35690163

RESUMO

Burkholderia sacchari LFM101 LMG19450T is a Brazilian bacterium isolated from sugarcane crops soil and a promising biotechnological platform for bioprocesses. It is an efficient producer of poly(3-hydroxybutyrate) from carbohydrates including xylose. In the present work, the expression of B. sacchari xylose consumption genes (xylA, xylB and tktA) was combined with the expression of Aeromonas sp. phaC (PHA synthase), aiming to increase both the growth rates in xylose and the 3-hydroxyhexanoate (3HHx) molar fractions in the produced PHA. Genes were cloned into pBBR1MCS-2 vectors and then expressed in the B. sacchari PHA- mutant LFM344. Maximum specific growth rates on xylose and PHA accumulation capacity of all recombinants were evaluated. In bioreactor experiments, up to 55.5 % CDW was accumulated as copolymer, hexanoate conversion to 3HHx raised from 2 % to 54 % of the maximum theoretical value, compared to wild type. 3HHx mol% ranged from 8 to 35, and molecular weights were between 111 and 220 kg/mol. Thermal analysis measurement showed a decrease in Tg and Tm values with higher 3HHx fraction, indicating improved thermomechanical characteristics. Recombinants construction and bioreactor strategies allowed the production of P(3HB-co-3HHx) with controlled monomeric composition from xylose and hexanoate, allowing its application in diverse fields, including the medical area.


Assuntos
Caproatos , Xilose , Ácido 3-Hidroxibutírico , Burkholderiaceae , Hidroxibutiratos/metabolismo
13.
J Ethnopharmacol ; 295: 115408, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-35659565

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Sargassum fusiforme (Harvey) Setchell, or Haizao, has been used in traditional Chinese medicine (TCM) since at least the eighth century a.d. S. fusiforme is an essential component of several Chinese formulas, including Haizao Yuhu Decoction, used to treat goiter, and Neixiao Lei Li Wan used to treat scrofuloderma. The pharmacological efficacy of S. fusiforme may be related to its anti-inflammatory effect. AIM OF THE STUDY: To determine the structural characteristics of SFF-32, a fucoidan fraction from S. fusiforme, and its antagonistic effect against P-selectin mediated function. MATERIALS AND METHODS: The primary structure of SFF-32 was determined using methylation/GC-MS and NMR analysis. Surface morphology and solution conformation of SFF-32 were determined by scanning electron microscopy (SEM), Congo red test, and circular dichroic (CD) chromatography, respectively. The inhibitory effects of SFF-32 against the binding of P-selectin to HL-60 cells were evaluated using flow cytometry, static adhesion assay, and parallel-plate flow chamber assay. Furthermore, the blocking effect of SFF-32 on the interaction between P-selectin and PSGL-1 was evaluated using an in vitro protein binding assay. RESULTS: The main linkage types of SFF-32 were proven to →[3)-α-l-Fucp-(1→3,4)-α-l-Fucp-(1]2→[4)-ß-d-Manp-(1→3)-d-GlcAp-(1]2→4)-ß-d-Manp-(1→3)-ß-d-Glcp-(1→4)-ß-d-Manp-(1→2,3)-ß-d-Galp-(1→4)-ß-d-Manp-(1→[4)-α-l-Rhap-(1]3→. The sulfated unit or terminal xylose residues were attached to the backbone through the C-3 of some fucose residues and terminal xylose residues were attached to C-3 of galactose residues. Moreover, SFF-32 disrupted P-selectin-mediated cell adhesion and rolling as well as blocked the interaction between P-selectin and its physiological ligand PSGL-1 in a dose-dependent manner. CONCLUSIONS: Blocking the binding between P-selectin and PSGL-1 is the possible underlying mechanism by which SFF-32 inhibits P-selectin-mediated function, which demonstrated that SFF-32 may be a potential anti-inflammatory lead compound.


Assuntos
Sargassum , Anti-Inflamatórios , Humanos , Selectina-P/metabolismo , Polissacarídeos/química , Polissacarídeos/farmacologia , Sargassum/química , Xilose
14.
Bioorg Chem ; 126: 105887, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35661527

RESUMO

Nowadays, alkali-tolerant ß-xylosidases and their molecular mechanism of pH adaptability have been poorly studied. Here, a novel GH43 ß-xylosidase (XYLO) was isolated from Bacillus clausii TCCC 11004, and the recombinant ß-xylosidase (rXYLO) was most active at pH 8.0 and stable in a broad pH range (7.0-11.0), exhibiting superior alkali tolerance. Molecular dynamics simulation indicated that XYLO showed a notable overall structural stability and an enlargement of substrate binding pocket under alkaline condition, resulting in the formation of a new hydrogen bond between substrate and Arg286 of XYLO, and the tight binding played a key role in improving the XYLO activity with the increasing pH. Moreover, rXYLO with an endo-xylanase resulted in high xylose yields by hydrolyzing alkali-extracted xylan from agricultural wastes. This work would provide an alkali-tolerant ß-xylosidase, enhance the understanding for the relationship of structure and activity adapted to the high-alkaline environment, and promote its application in xylose production.


Assuntos
Bacillus clausii , Xilosidases , Álcalis , Bacillus clausii/metabolismo , Concentração de Íons de Hidrogênio , Especificidade por Substrato , Xilose/metabolismo , Xilosidases/química
15.
Appl Microbiol Biotechnol ; 106(12): 4587-4606, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35708749

RESUMO

The transport of substrates across the cell membrane plays an essential role in nutrient assimilation by yeasts. The establishment of an efficient microbial cell factory, based on the maximum use of available carbon sources, can generate new technologies that allow the full use of lignocellulosic constituents. These technologies are of interest because they could promote the formation of added-value products with economic feasibility. In silico analyses were performed to investigate gene sequences capable of encoding xylose transporter proteins in the Candida tropicalis genome. The current study identified 11 putative transport proteins that have not yet been functionally characterized. A phylogenetic tree highlighted the potential C. tropicalis xylose-transporter proteins CtXUT1, CtXUT4, CtSTL1, CtSTL2, and CtGXT2, which were homologous to previously characterized and reported xylose transporters. Their expression was quantified through real-time qPCR at defined times, determined through a kinetic analysis of the microbial growth curve in the absence/presence of glucose supplemented with xylose as the main carbon source. The results indicated different mRNA expression levels for each gene. CtXUT1 mRNA expression was only found in the absence of glucose in the medium. Maximum CtXUT1 expression was observed in intervals of the highest xylose consumption (21 to 36 h) that corresponded to consumption rates of 1.02 and 0.82 g/L/h in the formulated media, with xylose as the only carbon source and with glucose addition. These observations indicate that CtXUT1 is an important xylose transporter in C. tropicalis. KEY POINTS: • Putative xylose transporter proteins were identified in Candida tropicalis; • The glucose concentration in the cultivation medium plays a key role in xylose transporter regulation; • The transporter gene CtXUT1 has an important role in xylose consumption by Candida tropicalis.


Assuntos
Candida tropicalis , Xilose , Candida tropicalis/genética , Candida tropicalis/metabolismo , Carbono/metabolismo , Proteínas de Transporte/genética , Biologia Computacional , Fermentação , Expressão Gênica , Glucose/metabolismo , Cinética , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Pentoses/metabolismo , Filogenia , RNA Mensageiro/metabolismo , Xilitol , Xilose/metabolismo
16.
Bioresour Technol ; 359: 127453, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35700903

RESUMO

Low-temperature biorefineries inhibit the multiplication of undesired microorganisms, improve product purity and reduce economic costs. Herein, to improve the 2,3-butanediol (2,3-BD) bioconversion efficiency from hemicellulose, a psychrotrophic hemicellulose-degrading strain Raoultella terrigena HC6 with high ß-xylosidase activity 1520 U/mL was isolated and genetically modified. Xylan (hemicellulose replacement) was depolymerized into xylooligosaccharides (XOS) and xylose by HC6, which were further converted into 2,3-BD. Transcriptomic analysis revealed that ß-xylosidase gene (xynB) and xylose isomerase gene (xylA), which are beneficial for increasing the carbon flux from xylan to 2,3-BD, were significantly upregulated 56.9-fold and 234-fold, respectively. A recombinant strain was constructed by overexpressing xynB in HC6, which obtained 0.389 g/g yield of 2,3-BD from hemicellulose extracted from corn straw at 15 °C. This study proposed a promised strategy for the bioconversion of agricultural waste into 2,3-BD at low temperatures and provides a basis for future efforts in the achievement of carbon neutrality.


Assuntos
Butileno Glicóis , Xilanos , Enterobacteriaceae , Polissacarídeos , Xilose
17.
Bioresour Technol ; 359: 127502, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35724907

RESUMO

A choline chloride-formic acid (ChCl-FA) pretreatment followed by enzymatic hydrolysis and fermentation were developed in this work for co-produce bioethanol, xylose, and lignin from eucalyptus. Results showed that ChCl-FA pretreatment can simultaneously degrade the xylan (∼95.2%) and lignin (∼74.4%) in eucalyptus, and obtained the pretreated eucalyptus having high glucan content and a numbers of cracks and holes, which was conducive to follow-up cellulase attacking. The hydrolysis experiments showed the maximum yield of glucose of 100 g eucalyptus was 35.3 g, which was equivalent to 90.3% of glucan in eucalyptus feedstock. The fermentation of enzymatic hydrolysate finally achieved the ethanol yield of 16.5 g, which corresponded to 74.5% theoretical ethanol yield from initial glucan in eucalyptus. In addition, 12.1 g xylose and 23.9 g lignin also could be obtained in pretreated liquid or/and hydrolysis residue, which represented for 61.4% xylan and 80.7% lignin in eucalyptus feedstock, respectively.


Assuntos
Eucalyptus , Xilose , Colina , Etanol/metabolismo , Eucalyptus/química , Fermentação , Formiatos , Glucanos/metabolismo , Hidrólise , Lignina/química , Xilanos/química , Xilose/metabolismo
18.
Microb Cell Fact ; 21(1): 105, 2022 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-35643525

RESUMO

BACKGROUND: Various inhibitors coexist in the hydrolysate derived from lignocellulosic biomass. They inhibit the performance of Saccharomyces cerevisiae and further restrict the development of industrial bioethanol production. Transcription factors are regarded as targets for constructing robust S. cerevisiae by genetic engineering. The tolerance-related transcription factors have been successively reported, while their regulatory mechanisms are not clear. In this study, we revealed the regulation mechanisms of Haa1p and Tye7p that had outstanding contributions to the improvement of the fermentation performance and multiple inhibitor tolerance of S. cerevisiae. RESULTS: Comparative transcriptomic analyses were applied to reveal the regulatory mechanisms of Haa1p and Tye7p under mixed sugar fermentation conditions with mixed inhibitors [acetic acid and furfural (AFur)] or without inhibitor (C) using the original strain s6 (S), the HAA1-overexpressing strain s6H3 (H), and the TYE7-overexpressing strain s6T3 (T). The expression of the pathways related to carbohydrate, amino acid, transcription, translation, cofactors, and vitamins metabolism was enhanced in the strains s6H3 and s6T3. Compared to C_H vs. C_S group, the unique DEGs in AFur_H vs. AFur_S group were further involved in oxidative phosphorylation, purine metabolism, vitamin B6 metabolism, and spliceosome under the regulation of Haa1p. A similar pattern appeared under the regulation of Tye7p, and the unique DEGs in AFur_T vs. AFur_S group were also involved in riboflavin metabolism and spliceosome. The most significant difference between the regulations of Haa1p and Tye7p was the intracellular energy supply. Haa1p preferred to enhance oxidative phosphorylation, while Tye7p tended to upregulate glycolysis/gluconeogenesis. CONCLUSIONS: Global gene expressions could be rewired with the overexpression of HAA1 or TYE7. The positive perturbations of energy and amino acid metabolism were beneficial to the improvement of the fermentation performance of the strain. Furthermore, strengthening of key cofactor metabolism, and transcriptional and translational regulation were helpful in improving the strain tolerance. This work provides a novel and comprehensive understanding of the regulation mechanisms of Haa1p and Tye7p in S. cerevisiae.


Assuntos
Proteínas de Saccharomyces cerevisiae , Xilose , Ácidos/metabolismo , Aminoácidos/metabolismo , Furaldeído/metabolismo , Glucose/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Transativadores/metabolismo , Fatores de Transcrição/genética , Xilose/metabolismo
19.
J Hazard Mater ; 437: 129345, 2022 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-35716565

RESUMO

Reducing the harm of heavy metals to the environment has been a major scientific challenge. In this study, D-(+)-xylose was used to prepare an adsorbent with rich O groups and three-dimensional porous structures for Cr(VI) adsorption. What's more, the adsorption sites of many oxygen groups in the material were combined with the three-dimensionally connected porous structures, which made the adsorption sites fully in contact with Cr(VI). At the concentration of 300 mg/L, the removal rate of Cr(VI) was 94.50%, 6.4 times that of the non-porous treatment and 9.6 times that of the non-porous and O group treatment. The adsorbent showed a high adsorption capacity (910.10 mg/g) for Cr(VI), and the adsorption model proved that the adsorbent was a multi-molecular layer adsorbent. In addition, the adsorption was controlled by chemical reaction and diffusion, which was also attributed to the three-dimensional porous structure and abundant oxygen groups of the material. XPS and FTIR indicated that four O groups participated in the adsorption reaction (-OH, C-O-C, CO, and C-O), and C-O-C and C-O were the main reaction sites. After treating wastewater from electroplating plants with X-PC, the discharged water met international and domestic discharge standards (Cr(VI) removal rate> 99.90%). This work provides a new idea for the application of sugars in the environment and the design of porous adsorbents.


Assuntos
Águas Residuárias , Poluentes Químicos da Água , Adsorção , Cromo/química , Concentração de Íons de Hidrogênio , Cinética , Oxigênio , Águas Residuárias/química , Poluentes Químicos da Água/química , Xilose
20.
Chemosphere ; 305: 135417, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35750228

RESUMO

Chlorine disinfection is widely applied in drinking water treatment plant to inactivate pathogens in drinking water, but it unintentionally reacts with organic matter present in source waters and generates halogenated disinfection byproducts (DBPs). Sugar is one of the most commonly used seasoning in our diet. The addition of sugar could significantly improve the taste of the beverages; however, the effects of sugar on DBP formation and transformation remain unknown. In this study, the effects of sugar type and dose on the halogenated DBP formation in chlorinated boiled real tap water were evaluated during making hot beverages. We found that sugar can react with chlorine residual in tap water and generate halogenated DBPs. As the most commonly used table sugar, the addition of sucrose in the water sample at 100 or 500 mg/L as C could increase the level of total organic halogen (TOX) by ∼35%, when compared with the boiled tap water sample without sugar addition. In addition, fifteen reported and new polar brominated and chlorinated DBPs were detected and proposed from the reaction between chlorine and sucrose; accordingly, the corresponding transformation pathways were also proposed. Moreover, the DBP formation in the chlorinated boiled real tap water samples with the addition of xylose, glucose, sucrose, maltose and lactose were also investigated. By comparing with the TOX levels in the water samples with different sugar addition and their calculated TOX risk indexes, it was suggested that applying xylose as a sweetener in beverages could not only obtain a relatively high sweetness but also minimize the adverse effect inducing by halogenated DBPs during making hot beverages.


Assuntos
Desinfetantes , Água Potável , Poluentes Químicos da Água , Purificação da Água , Cloro , Desinfetantes/análise , Desinfecção , Halogenação , Halogênios , Sacarose , Açúcares , Poluentes Químicos da Água/análise , Xilose
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