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1.
BMC Plant Biol ; 23(1): 44, 2023 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-36658483

RESUMO

BACKGROUND: Testa color is an important trait of peanut (Arachis hypogaea L.) which is closely related with the nutritional and commercial value. Pink and red are main color of peanut testa. However, the genetic mechanism of testa color regulation in peanut is not fully understood. To elucidate a clear picture of peanut testa regulatory model, samples of pink cultivar (Y9102), red cultivar (ZH12), and two RNA pools (bulk red and bulk pink) constructed from F4 lines of Y9102 x ZH12 were compared through a bulk RNA-seq approach. RESULTS: A total of 2992 differential expressed genes (DEGs) were identified among which 317 and 1334 were up-regulated and 225 and 1116 were down-regulated in the bulk red-vs-bulk pink RNA pools and Y9102-vs-ZH12, respectively. KEGG analysis indicates that these genes were divided into significantly enriched metabolic pathways including phenylpropanoid, flavonoid/anthocyanin, isoflavonoid and lignin biosynthetic pathways. Notably, the expression of the anthocyanin upstream regulatory genes PAL, CHS, and CHI was upregulated in pink and red testa peanuts, indicating that their regulation may occur before to the advent of testa pigmentation. However, the differential expression of down-stream regulatory genes including F3H, DFR, and ANS revealed that deepening of testa color not only depends on their gene expression bias, but also linked with FLS inhibition. In addition, the down-regulation of HCT, IFS, HID, 7-IOMT, and I2'H genes provided an alternative mechanism for promoting anthocyanin accumulation via perturbation of lignin and isoflavone pathways. Furthermore, the co-expression module of MYB, bHLH, and WRKY transcription factors also suggested a fascinating transcriptional activation complex, where MYB-bHLH could utilize WRKY as a co-option during the testa color regulation by augmenting anthocyanin biosynthesis in peanut. CONCLUSIONS: These findings reveal candidate functional genes and potential strategies for the manipulation of anthocyanin biosynthesis to improve peanut varieties with desirable testa color.


Assuntos
Antocianinas , Arachis , Antocianinas/metabolismo , Arachis/genética , Arachis/metabolismo , Redes Reguladoras de Genes , Lignina/metabolismo , Pigmentação/genética , Regulação da Expressão Gênica de Plantas , Cor , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Perfilação da Expressão Gênica
2.
Nutrients ; 15(1)2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36615876

RESUMO

Lignin-carbohydrate complexes (LCCs) are emerging as a new and natural product with pharmacological and nutraceutical potential. It is uncertain, however, whether LCCs have a positive effect on the microbiota of the gut based on the current evidence. Here, the LCC extracted from beechwood (BW-LCC) was used as a substrate for in vitro fermentation. The lignin in BW-LCC consisted of guaiacyl (G) and syringyl (S) units, which are mainly linked by ß-O-4 bonds. After 24 h of in vitro fermentation, the pH had evidently declined. The concentrations of acetic acid and propionic acid, the two main short-chain fatty acids (SCFAs), were significantly higher than in the control group (CK). In addition, BW-LCC altered the microbial diversity and composition of gut microbes, including a reduction in the relative abundance of Firmicutes and an increase in the relative abundance of Proteobacteria and Bacteroidetes. The relative abundance of Escherichia coli-Shigella and Bacteroides were the most variable at the genus level. The genes of carbohydrate-active enzymes (CAZymes) also changed significantly with the fermentation and were related to the changes in microbes. Notably, the auxiliary actives (AAs), especially AA1, AA2, and AA3_2, play important roles in lignin degradation and were significantly enriched and concentrated in Proteobacteria. From this study, we are able to provide new perspectives on how gut microbes utilize LCC.


Assuntos
Carboidratos , Lignina , Lignina/química , Lignina/metabolismo , Fermentação , Carboidratos/química , Bactérias/metabolismo , Ácidos Graxos Voláteis/metabolismo , Proteobactérias/metabolismo
3.
Environ Sci Technol ; 57(2): 1144-1156, 2023 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-36599031

RESUMO

Conventional phototrophic cultivation for microalgae production suffers from low and unstable biomass productivity due to limited and unreliable light transmission outdoors. Alternatively, the use of a renewable lignocellulose-derived carbon source, cellulosic hydrolysate, offers a cost-effective and sustainable pathway to cultivate microalgae heterotrophically with high algal growth rate and terminal density. In this study, we evaluate the feasibility of cellulosic hydrolysate-mediated heterotrophic cultivation (Cel-HC) for microalgae production by performing economic and environmental comparisons with phototrophic cultivation through techno-economic analysis and life cycle assessment. We estimate a minimum selling price (MSP) of 4722 USD/t for producing high-purity microalgae through Cel-HC considering annual biomass productivity of 300 t (dry weight), which is competitive with the conventional phototrophic raceway pond system. Revenues from the lignocellulose-derived co-products, xylose and fulvic acid fertilizer, could further reduce the MSP to 2976 USD/t, highlighting the advantages of simultaneously producing high-value products and biofuels in an integrated biorefinery scheme. Further, Cel-HC exhibits lower environmental impacts, such as cumulative energy demand and greenhouse gas emissions, than phototrophic systems, revealing its potential to reduce the carbon intensity of algae-derived commodities. Our results demonstrate the economic and environmental competitiveness of heterotrophic microalgae production based on renewable bio-feedstock of lignocellulose.


Assuntos
Microalgas , Microalgas/metabolismo , Lignina/metabolismo , Biocombustíveis , Biomassa , Carbono/metabolismo
4.
Appl Microbiol Biotechnol ; 107(2-3): 535-542, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36607404

RESUMO

Studies on the chemical mechanisms of furfural formation showed the possibility to apply a new differential catalysis of hemicellulose - its depolymerisation and pentose dehydration to furfural. This change led to the increase in furfural yield and essential decrease of cellulose destruction. The lignocellulose residue that remains after the production of furfural may be subjected to enzymatic hydrolysis to glucose and the subsequent fermentation to ethanol. The remaining lignin appeared to be suitable for the production of additional various value-added products including medicinal mushrooms and laccase-containing enzyme complexes. Based on these developments, an innovative concept is proposed for the waste-free use of lignocellulose to obtain various valuable products. KEY POINTS: • New chemical mechanism of furfural production. • New lignocellulose pretreatment does not damage cellulose and lignin. • Lignocellulose may be processed using waste-free technology.


Assuntos
Etanol , Lignina , Lignina/metabolismo , Furaldeído , Celulose , Hidrólise , Fermentação
5.
Proc Natl Acad Sci U S A ; 120(4): e2212246120, 2023 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-36652470

RESUMO

Lignin valorization is being intensely pursued via tandem catalytic depolymerization and biological funneling to produce single products. In many lignin depolymerization processes, aromatic dimers and oligomers linked by carbon-carbon bonds remain intact, necessitating the development of enzymes capable of cleaving these compounds to monomers. Recently, the catabolism of erythro-1,2-diguaiacylpropane-1,3-diol (erythro-DGPD), a ring-opened lignin-derived ß-1 dimer, was reported in Novosphingobium aromaticivorans. The first enzyme in this pathway, LdpA (formerly LsdE), is a member of the nuclear transport factor 2 (NTF-2)-like structural superfamily that converts erythro-DGPD to lignostilbene through a heretofore unknown mechanism. In this study, we performed biochemical, structural, and mechanistic characterization of the N. aromaticivorans LdpA and another homolog identified in Sphingobium sp. SYK-6, for which activity was confirmed in vivo. For both enzymes, we first demonstrated that formaldehyde is the C1 reaction product, and we further demonstrated that both enantiomers of erythro-DGPD were transformed simultaneously, suggesting that LdpA, while diastereomerically specific, lacks enantioselectivity. We also show that LdpA is subject to a severe competitive product inhibition by lignostilbene. Three-dimensional structures of LdpA were determined using X-ray crystallography, including substrate-bound complexes, revealing several residues that were shown to be catalytically essential. We used density functional theory to validate a proposed mechanism that proceeds via dehydroxylation and formation of a quinone methide intermediate that serves as an electron sink for the ensuing deformylation. Overall, this study expands the range of chemistry catalyzed by the NTF-2-like protein family to a prevalent lignin dimer through a cofactorless deformylation reaction.


Assuntos
Liases , Lignina/metabolismo , Proteínas de Bactérias/metabolismo , Oxirredutases/metabolismo , Estereoisomerismo
6.
Food Chem ; 409: 135297, 2023 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-36623356

RESUMO

Segment drying is a common internal physiological disorder in citrus fruit, and vesicles get granulated or collapsed. This study aimed to probe whether and how the phenylpropanoid metabolism changes in vesicles during collapse of blood orange (Citrus sinensis cv. Tarocco). Vesicle collapse led to a decrease in the content of nutrients and flavonoids, while an increase in lignin content. This disorder was further associated with the increasing enzyme activities and gene expression levels of both the general phenylpropanoid pathway and branch pathway of lignin synthesis, while decreasing enzyme activities and gene expression levels of branch pathway of flavonoids synthesis. Targeted metabolomics analysis of 14 metabolites of the lignin pathway revealed that lignin precursors were accumulated in collapsed vesicles. We provide solid evidence that phenylpropanoid metabolism could be activated, and, intriguingly, metabolic flux may be shuttled to lignin precursors synthesis rather than flavonoids synthesis in vesicles during collapse of blood orange.


Assuntos
Citrus sinensis , Citrus , Citrus sinensis/química , Lignina/metabolismo , Citrus/química , Flavonoides/análise , Metabolômica , Frutas/química
7.
Int J Mol Sci ; 24(2)2023 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-36674995

RESUMO

Lignin degradation is an effective means of achieving the high-value application of lignin, but degradation usually requires the use of high temperatures and harsh reaction-conditions. This study describes a green, mild approach for the degradation of lignin, in which chlorine dioxide (ClO2) was used for the oxidative degradation of lignin (IL) in an acidic aqueous suspension at room temperature. The optimal process conditions were: 30 mL of ClO2 solution (2.5 mg·L-1), pH 4.5 and 3 h. The FT-IR, NMR (1H NMR, 2D-HSQC and 31P NMR), XPS and GPC analyses indicated that lignin could be degraded by ClO2 relatively well at room temperature, to form quinones and muconic acids. Additionally, DIL was reduced to substances with a high phenolic-hydroxyl (OH) content (RDIL) under the presence of NaBH4, which further confirmed the composition of DIL and which can be applied to the development of lignin-based phenolic resins, providing a reference for the further modification as well as the utilization of DIL.


Assuntos
Lignina , Óxidos , Lignina/metabolismo , Temperatura , Espectroscopia de Infravermelho com Transformada de Fourier , Cloro
8.
Int J Mol Sci ; 24(2)2023 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-36675053

RESUMO

Asparagus belongs to the Liliaceae family and has important economic and pharmacological value. Lignin plays a crucial role in cell wall structural integrity, stem strength, water transport, mechanical support and plant resistance to pathogens. In this study, various biological methods were used to study the mechanism of shading on the asparagus lignin accumulation pathway. The physiological results showed that shading significantly reduced stem diameter and cell wall lignin content. Microstructure observation showed that shading reduced the number of vascular bundles and xylem area, resulting in decreased lignin content, and thus reducing the lignification of asparagus. Cinnamic acid, caffeic acid, ferulic acid and sinapyl alcohol are crucial intermediate metabolites in the process of lignin synthesis. Metabolomic profiling showed that shading significantly reduced the contents of cinnamic acid, caffeic acid, ferulic acid and sinapyl alcohol. Transcriptome profiling identified 37 differentially expressed genes related to lignin, including PAL, C4H, 4CL, CAD, CCR, POD, CCoAOMT, and F5H related enzyme activity regulation genes. The expression levels of POD, CCoAOMT, and CCR genes were significantly decreased under shading treatment, while the expression levels of CAD and F5H genes exhibited no significant difference with increased shading. The downregulation of POD, CCoAOMT genes and the decrease in CCR gene expression levels inhibited the activities of the corresponding enzymes under shading treatment, resulting in decreased downstream content of caffeic acid, ferulic acid, sinaperol, chlorogenic acid and coniferin. A significant decrease in upstream cinnamic acid content was observed with shading, which also led to decreased downstream metabolites and reduced asparagus lignin content. In this study, transcriptomic and metabolomic analysis revealed the key regulatory genes and metabolites of asparagus lignin under shading treatment. This study provides a reference for further understanding the mechanism of lignin biosynthesis and the interaction of related genes.


Assuntos
Lignina , Transcriptoma , Lignina/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas
9.
Bioresour Technol ; 370: 128526, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36572161

RESUMO

Lignin degradation represents a significant challenge in biological valorization, but it is suffering from insufficiency, putting barriers to efficient lignin conversion. Herein, the study first develops a highly efficient laccase secretion apparatus, enabling high enzyme activity of 184 U/mL, complementing the biochemical limits on lignin depolymerization well in Halomonas sp. Y3. Further engineering of PHA biosynthesis produces a significantly high PHA titer of 286, 742, and 868 mg/L from alkaline lignin, catechol, and protocatechuate, respectively. The integration of laccase-secretion and PHA production modules enables a record titer of 693 and 1209 mg/L in converting lignin and lignin-containing stream to PHA, respectively. The titer is improved furtherly to 740 and 1314 mg/L by developing a non-sterilized fermentation. This study advances a cheaper and greener production of valuable chemicals from lignin by constructing a biosynthetic platform for PHA production and provides novel insight into the lignin conversion by extremophilic microbes.


Assuntos
Halomonas , Poli-Hidroxialcanoatos , Lignina/metabolismo , Halomonas/genética , Halomonas/metabolismo , Lacase , Engenharia Metabólica
10.
Biotechnol Lett ; 45(1): 95-104, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36482053

RESUMO

OBJECTIVES: Maximizing the utility value of enzymes was achieved by exploring the effects of small molecules on the efficiency of lignin degradation by lignin peroxidase. METHODS: Using wheat straw as raw material and taking lignin degradation rate as index, it was found that laccase, glucose oxidase, malonic acid, citric acid, ZnSO4, CaCl2 could promote the lignin degradation by the lignin peroxidase from Irpex lacteus, respectively. Moreover, glucose oxidase, malonic acid and CaCl2 had obvious synergy effects on lignin degradation by the lignin peroxidase. RESULTS: The optimal conditions of lignin degradation were obtained by response surface experiment: 4% glucose oxidase, 0.74% malonic acid and 3.29% CaCl2 were added for synergistic degradation at 37 â„ƒ with 50% of water content. After 72 h quickly enzymatic hydrolysis, the degradation rate of lignin was 45.84%. CONCLUSIONS: A new green and efficient method for lignin removal from straw was obtained, which provided a reference for the efficient utilization of straw and lignin peroxidase.


Assuntos
Glucose Oxidase , Lignina , Lignina/metabolismo , Cloreto de Cálcio
11.
New Phytol ; 237(1): 251-264, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36196006

RESUMO

Ester-linked p-coumarate (pCA) is a hallmark feature of the secondary cell walls in commelinid monocot plants. It has been shown that pCA groups arise during lignin polymerisation from the participation of monolignol conjugates assembled by p-coumaroyl-CoA:monolignol transferase (PMT) enzymes, members of the BAHD superfamily of acyltransferases. Herein, we report that a eudicot species, kenaf (Hibiscus cannabinus), naturally contains p-coumaroylated lignin in the core tissues of the stems but not in the bast fibres. Moreover, we identified a novel acyltransferase, HcPMT, that shares <30% amino acid identity with known monocot PMT sequences. Recombinant HcPMT showed a preference in enzyme assays for p-coumaroyl-CoA and benzoyl-CoA as acyl donor substrates and sinapyl alcohol as an acyl acceptor. Heterologous expression of HcPMT in hybrid poplar trees led to the incorporation of pCA in lignin, but no improvement in the saccharification potential of the wood. This work illustrates the value in mining diverse plant taxa for new monolignol acyltransferases. Furthermore, the occurrence of pCA outside monocot lineages may represent another example of convergent evolution in lignin structure. This discovery expands textbook views on cell wall biochemistry and provides a new molecular tool for engineering the lignin of biomass feedstock plants.


Assuntos
Lignina , Populus , Lignina/metabolismo , Parede Celular/metabolismo , Aciltransferases/metabolismo , Populus/metabolismo , Coenzima A/análise , Coenzima A/metabolismo
12.
Appl Microbiol Biotechnol ; 107(1): 273-286, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36477928

RESUMO

Thermoalkaliphilic laccase (CtLac) from the Caldalkalibacillus thermarum strain TA2.A1 has advantageous properties with potential industrial applications, such as high enzyme activity and stability at 70 °C and pH 8.0. In the present study, a directed evolution approach using a combination of random and site-directed mutagenesis was adopted to enhance the laccase activity of CtLac. Spectrophotometric assay and real-time oxygen measurement techniques were employed to compare and evaluate the enzyme activity among mutants. V243 was targeted for site-directed mutagenesis based on library screening. V243D showed a 25-35% higher laccase activity than wild-type CtLac in the spectrophotometric assay and oxygen consumption measurement results. V243D also showed higher catalytic efficiency than wild-type CtLac with decreased Km and increased kcat values. In addition, V243D enhanced oxidative degradation of the lignin model compound, guaiacylglycerol-ß-guaiacyl ether (GGGE), by 10% and produced a 5-30% increase in high-value aldehydes than wild-type CtLac under optimal enzymatic conditions (i.e., 70 °C and pH 8.0). Considering the lack of protein structural information, we used the directed evolution approach to predict Val at the 243 position of CtLac as one of the critical amino acids contributing to the catalytic efficiency of the enzyme. Moreover, it found that the real-time oxygen measurement technique could overcome the limitations of the spectrophotometric assay, and apply to evaluate oxidase activity in mutagenesis research. KEY POINTS: • CtLac was engineered for enhanced laccase activity through directed evolution approach • V243D showed higher catalytic efficiency (kcat/Km) than wild-type CtLac • V243D produced higher amounts of high-value aldehydes from rice straw than wild-type CtLac.


Assuntos
Lacase , Lignina , Lacase/metabolismo , Lignina/metabolismo , Mutagênese Sítio-Dirigida , Aldeídos , Oxigênio
13.
J Plant Physiol ; 280: 153900, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36525838

RESUMO

Although several aspects of lignin metabolism have been extensively characterized, the mechanism(s) by which lignin monomers are transported across the plasma membrane remains largely unknown. Biochemical, proteomic, expression and co-expression analyses from several plant species support the involvement of active transporters, mainly those belonging to the ABC superfamily. Here, we report on the genome-wide characterization of the ABCG gene subfamily in the model C4 grass Setaria viridis and further identification of the members potentially involved in monolignol transport. A total of 48 genes encoding SvABCGs were found in the S. viridis genome, from which 21 SvABCGs were classified as full-size transporters and 27 as half-size transporters. Comprehensive analysis of the ABCG subfamily in S. viridis based on expression and co-expression analyses support a role for SvABCG17 in monolignol transport: (i) SvABCG17 is orthologous to AtABCG29, a monolignol transporter in Arabidopsis thaliana; (ii) SvABCG17 displays a similar expression profile to that of lignin biosynthetic genes in a set of different S. viridis tissues and along the elongating internode; (iii) SvABCG17 is highly co-expressed with lignin-related genes in a public transcriptomic database; (iv) SvABCG17displays particularly high expression in the top of the S. viridis elongating internode, a tissue undergoing active lignification; (v) SvABCG17 mRNA localization coincides with the histochemical pattern of lignin deposition; and (vi) the promoter of SvABCG17 is activated by secondary cell wall-associated transcription factors, especially by lignin-specific activators of the MYB family. Further studies might reveal further aspects of this potential monolignol transporter, including its real substrate specificity and whether it works redundantly with other ABC members during S. viridis lignification.


Assuntos
Arabidopsis , Setaria (Planta) , Lignina/metabolismo , Setaria (Planta)/genética , Proteômica , Proteínas de Membrana Transportadoras/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo
14.
Int J Biol Macromol ; 228: 647-658, 2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36584775

RESUMO

In this study, Lentinula edodes and Pleurotus ostreatus were selected as representatives to explore the effects of corn straw and xylosma sawdust on the production and activity of polysaccharides by edible fungi during solid-state fermentation. The results clearly indicated that the addition of lignocellulose could promote the polysaccharide content compared to the control group. Meanwhile, the hydroxyl radical scavenging activity of polysaccharides reached the maximum when the glucose concentration was 1.5 %, and among them, the xylosma sawdust groups for two fungi (L. edodes-32.37 %, P. ostreatus-25.86 %) both performed better than corn straw groups (L. edodes-24.96 %, P. ostreatus-20.80 %). In addition, structural characterization and degradation mode analysis were carried out. The results showed that the structure of the xylosma had a stronger destruction than corn straw. The activities of lignocellulolytic enzymes such as carboxymethyl cellulase, filter paper enzyme, ß-glucosidase and xylanase increased rapidly in the early stage of fermentation, which could degrade the cellulose and hemicellulose as raw materials for the synthesis of active polysaccharides.


Assuntos
Pleurotus , Cogumelos Shiitake , Pleurotus/metabolismo , Fermentação , Zea mays/metabolismo , Lignina/metabolismo , Polissacarídeos/metabolismo
15.
Bioresour Technol ; 370: 128510, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36538959

RESUMO

To explore the interaction mechanism of pseudo-lignin (PL) with cellulase and its influence on cellulose hydrolysis, different PLs were extracted from pretreated bamboo holocellulose (HC) using different organic solvents. Meanwhile, the real-time interaction of PL and cellulase was analyzed using surface plasmon resonance (SPR). The results showed that the extraction effect of the tetrahydrofuran and 1, 4-dioxane/water solution on PL was more effective than the ethanol/water solution. The inhibition of PL fraction obtained from HC by acid pretreatment with higher temperature showed less effect on Avicel's enzymatic hydrolysis. SPR analysis revealed that PL formed at higher pretreatment temperature had a lower dissociation rate after adsorption with cellulase. Besides, the binding affinity of PL (160 °C) to cellulase was much greater than that of PL obtained from 180 °C, indicating PL extracted at higher temperature treated biomass is more easily dissociated from cellulase after binding.


Assuntos
Celulase , Celulases , Celulose/metabolismo , Lignina/metabolismo , Ressonância de Plasmônio de Superfície , Hidrólise , Água , Celulase/metabolismo
16.
Metab Eng ; 75: 153-169, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36563956

RESUMO

Lignin displays a highly challenging renewable. To date, massive amounts of lignin, generated in lignocellulosic processing facilities, are for the most part merely burned due to lacking value-added alternatives. Aromatic lignin monomers of recognized relevance are in particular vanillin, and to a lesser extent vanillate, because they are accessible at high yield from softwood-lignin using industrially operated alkaline oxidative depolymerization. Here, we metabolically engineered C. glutamicum towards cis, cis-muconate (MA) production from these key aromatics. Starting from the previously created catechol-based producer C. glutamicum MA-2, systems metabolic engineering first discovered an unspecific aromatic aldehyde reductase that formed aromatic alcohols from vanillin, protocatechualdehyde, and p- hydroxybenzaldehyde, and was responsible for the conversion up to 57% of vanillin into vanillyl alcohol. The alcohol was not re-consumed by the microbe later, posing a strong drawback on the producer. The identification and subsequent elimination of the encoding fudC gene completely abolished vanillyl alcohol formation. Second, the initially weak flux through the native vanillin and vanillate metabolism was enhanced up to 2.9-fold by implementing synthetic pathway modules. Third, the most efficient protocatechuate decarboxylase AroY for conversion of the midstream pathway intermediate protocatechuate into catechol was identified out of several variants in native and codon optimized form and expressed together with the respective helper proteins. Fourth, the streamlined modules were all genomically combined which yielded the final strain MA-9. MA-9 produced bio-based MA from vanillin, vanillate, and seven structurally related aromatics at maximum selectivity. In addition, MA production from softwood-based vanillin, obtained through alkaline depolymerization, was demonstrated.


Assuntos
Corynebacterium glutamicum , Lignina , Lignina/metabolismo , Engenharia Metabólica , Corynebacterium glutamicum/genética , Corynebacterium glutamicum/metabolismo , Catecóis/metabolismo
17.
J Agric Food Chem ; 70(50): 15848-15858, 2022 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-36475725

RESUMO

Mycosporine-like amino acids (MAAs) have been used in cosmetics and pharmaceuticals. The purpose of this work was to develop yeast strains for sustainable and economical production of MAAs, especially shinorine. First, genes involved in MAA biosynthetic pathway from Actinosynnema mirum were introduced into Saccharomyces cerevisiae for heterologous shinorine production. Second, combinatorial expression of wild and mutant xylose reductase was adopted in the engineered S. cerevisiae to facilitate xylose utilization in the pentose phosphate pathway. Finally, the accumulation of sedoheptulose 7-phosphate (S7P) was attempted by deleting transaldolase-encoding TAL1 in the pentose phosphate pathway to increase carbon flux toward shinorine production. In fed-batch fermentation, the engineered strain (DXdT-M) produced 751 mg/L shinorine in 71 h. Ultimately, 54 mg/L MAAs was produced by DXdT-M from rice straw hydrolysate. The results suggest that shinorine production by S. cerevisiae might be a promising process for sustainable production and industrial applications.


Assuntos
Lignina , Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Biomassa , Lignina/metabolismo , Xilose/metabolismo , Fermentação
18.
BMC Plant Biol ; 22(1): 614, 2022 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-36575388

RESUMO

BACKGROUND: Uniconazole is an effective plant growth regulator that can be used in banana cultivation to promote dwarfing and enhance lodging resistance. However, the mechanisms underlying banana dwarfing induced by uniconazole are unknown. In uniconazole-treated bananas, gibberellin (GA) was downregulated compared to the control groups. An integrative analysis of transcriptomes and metabolomes was performed on dwarf bananas induced by uniconazole and control groups. The key pathways involved in uniconazole-induced dwarfism in banana were determined according to the overlap of KEGG annotation of differentially expressed genes and (DEGs) differential abundant metabolites (DAMs). RESULTS: Compared with the control groups, the levels of some flavonoids, tannins, and alkaloids increased, and those of most lipids, amino acids and derivatives, organic acids, nucleotides and derivatives, and terpenoids decreased in uniconazole-treated bananas. Metabolome analysis revealed the significant changes of flavonoids in uniconazole-treated bananas compared to control samples at both 15 days and 25 days post treatment. Transcriptome analysis shows that the DEGs between the treatment and control groups were related to a series of metabolic pathways, including lignin biosynthesis, phenylpropanoid metabolism, and peroxidase activity. Comprehensive analysis of the key pathways of co-enrichment of DEGs and DAMs from 15 d to 25 d after uniconazole treatment shows that flavonoid biosynthesis was upregulated. CONCLUSIONS: In addition to the decrease in GA, the increase in tannin procyanidin B1 may contribute to dwarfing of banana plants by inhibiting the activity of GA. The increased of flavonoid biosynthesis and the change of lignin biosynthesis may lead to dwarfing phenotype of banana plants. This study expands our understanding of the mechanisms underlying uniconazole-induced banana dwarfing.


Assuntos
Nanismo , Musa , Transcriptoma , Musa/genética , Musa/metabolismo , Lignina/metabolismo , Perfilação da Expressão Gênica , Flavonoides/metabolismo , Regulação da Expressão Gênica de Plantas
19.
PeerJ ; 10: e14426, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36523457

RESUMO

Agaricus bisporus growth alters the lignocellulosic composition and structure of compost. However, it is difficult to differentiate the enzyme activities of A. bisporus mycelia from the wider microbial community owing to the complication of completely speareting the mycelia from compost cultures. Macrogenomics analysis was employed in this study to examine the fermentation substrate of A. bisporus before and after mycelial growth, and the molecular mechanism of substrate utilization by A. bisporus mycelia was elucidated from the perspective of microbial communities and CAZymes in the substrate. The results showed that the relative abundance of A. bisporus mycelia increased by 77.57-fold after mycelial colonization, the laccase content was significantly increased and the lignin content was significantly decreased. Analysis of the CAZymes showed that AA10 family was extremely differentiated. Laccase-producing strains associated with AA10 family were mostly bacteria belonging to Thermobifida and Thermostaphylospora, suggesting that these bacteria may play a synergistic role in lignin decomposition along with A. bisporus mycelia. These findings provide preliminary evidence for the molecular mechanism of compost utilization by A. bisporus mycelia and offer a reference for the development and utilization of strains related to lignocellulose degradation.


Assuntos
Compostagem , Lignina , Lignina/metabolismo , Lacase/genética , Metagenômica
20.
Int J Mol Sci ; 23(22)2022 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-36430286

RESUMO

Extensive research efforts have been devoted to describing yeast alcohol oxidase (AO) and its promoter region, which is vastly applied in studies of heterologous gene expression. However, little is known about basidiomycetous AO and its physiological role in wood degradation. This review describes several alcohol oxidases from both white and brown rot fungi, highlighting their physicochemical and kinetic properties. Moreover, the review presents a detailed analysis of available AO-encoding gene promoter regions in basidiomycetous fungi with a discussion of the manipulations of culture conditions in relation to the modification of alcohol oxidase gene expression and changes in enzyme production. The analysis of reactions catalyzed by lignin-modifying enzymes (LME) and certain lignin auxiliary enzymes (LDA) elucidated the possible involvement of alcohol oxidase in the degradation of derivatives of this polymer. Combined data on lignin degradation pathways suggest that basidiomycetous AO is important in secondary reactions during lignin decomposition by wood degrading fungi. With numerous alcoholic substrates, the enzyme is probably engaged in a variety of catalytic reactions leading to the detoxification of compounds produced in lignin degradation processes and their utilization as a carbon source by fungal mycelium.


Assuntos
Basidiomycota , Madeira , Madeira/metabolismo , Lignina/metabolismo , Oxirredutases do Álcool/metabolismo , Fungos/metabolismo
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