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1.
Foods ; 13(12)2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38928896

RESUMO

The traditional Chinese Baijiu brewing process utilizes natural inoculation and open fermentation. The microbial composition and abundance in the microecology of Baijiu brewing often exhibit unstable characteristics, which directly results in fluctuations in Baijiu quality. The microbiota plays a crucial role in determining the quality of Baijiu. Analyzing the driving effect of technology and raw materials on microorganisms. Elucidating the source of core microorganisms and interactions between microorganisms, and finally utilizing single or multiple microorganisms to regulate and intensify the Baijiu fermentation process is an important way to achieve high efficiency and stability in the production of Baijiu. This paper provides a systematic review of the composition and sources of microbiota at different brewing stages. It also analyzes the relationship between raw materials, brewing processes, and brewing microbiota, as well as the steps involved in the implementation of brewing microbiota regulation strategies. In addition, this paper considers the feasibility of using Baijiu flavor as a guide for Baijiu brewing regulation by synthesizing the microbiota, and the challenges involved. This paper is a guide for flavor regulation and quality assurance of Baijiu and also suggests new research directions for regulatory strategies for other fermented foods.

2.
Talanta ; 276: 126282, 2024 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-38788382

RESUMO

Herein, spore@Cu-trimesic acid (TMA) biocomposites were prepared by self-assembling Cu-based metal-organic framework on the surface of Bacillus velezensis spores. The laccase-like activity of spore@Cu-TMA biocomposites was enhanced by 14.9 times compared with that of pure spores due to the reaction of Cu2+ ions with laccase on the spore surface and the microporous structure of Cu-TMA shell promoting material transport and increasing substrate accessibility. Spore@Cu-TMA rapidly oxidized and transformed 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) into ABTS●+ without using H2O2. Under optimum conditions, the ABTS●+ could be stored for 21 days at 4 °C and 7 days at 37 °C without the addition of any stabilizers, allowing for the large-scale preparation and long-term storage of ABTS●+. The ultrarobust stable ABTS●+ obtained with the use of Cu-TMA could effectively reduce the "back reaction" by preventing the leaching of the metabolites released by the spores. On the basis of these findings, a rapid, low-cost, and eco-friendly colorimetric platform was successfully developed for the detection of antioxidant capacity. Determination of antioxidant capacity for several antioxidants such as caffeic acid, glutathione, and Trolox revealed their corresponding limits of detection at 4.83, 8.89, and 7.39 nM, respectively, with linear ranges of 0.01-130, 0.01-140, and 0.01-180 µM, respectively. This study provides a facile way to prepare ultrarobust stable ABTS●+ and presents a potential application of spore@Cu-TMA biocomposites in food detection and bioanalysis.


Assuntos
Antioxidantes , Bacillus , Benzotiazóis , Cobre , Esporos Bacterianos , Ácidos Sulfônicos , Cobre/química , Ácidos Sulfônicos/química , Benzotiazóis/química , Antioxidantes/química , Antioxidantes/análise , Esporos Bacterianos/química , Bacillus/enzimologia , Lacase/química , Lacase/metabolismo , Estruturas Metalorgânicas/química , Ácidos Tricarboxílicos/química
3.
Food Chem ; 450: 139285, 2024 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-38631203

RESUMO

Theaflavins are beneficial to human health due to various bioactivities. Biosynthesis of theaflavins using polyphenol oxidase (PPO) is advantageous due to cost effectiveness and environmental friendliness. In this review, studies on the mechanism of theaflavins formation, the procedures to screen and prepare PPOs, optimization of reaction systems and immobilization of PPOs were described. The challenges associated with the mass biosynthesis of theaflavins, such as poor enzyme activity, undesirable subproducts and inclusion bodies of recombinant PPOs were presented. Further strategies to solve these challenges and improve theaflavins production, including enzyme engineering, immobilization enzyme technology, water-immiscible solvent-water biphasic systems and recombinant enzyme technology, were proposed.


Assuntos
Biflavonoides , Catequina , Catecol Oxidase , Biflavonoides/química , Biflavonoides/metabolismo , Biocatálise , Catequina/química , Catequina/metabolismo , Catecol Oxidase/metabolismo , Catecol Oxidase/química , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo
4.
Food Chem X ; 22: 101382, 2024 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-38665634

RESUMO

In this study, a fortified Daqu (FF Daqu) was prepared using high cellulase-producing Bacillus subtilis, and the effects of in situ fortification on the physicochemical properties, flavor, active microbial community and metabolism of Daqu were analyzed. The saccharification power, liquefaction power, and cellulase activity of the FF Daqu were significantly increased compared with that of the traditional Daqu (CT Daqu). The overall differences in flavor components and their contents were not significant, but the higher alcohols were lower in FF Daqu. The relative abundance of dominant active species in FF Daqu was 85.08% of the total active microbiota higher than 63.42% in CT Daqu, and the biomarkers were Paecilomyces variotii and Aspergillus cristatus, respectively. The enzymes related to starch and sucrose metabolic pathways were up-regulated and expressed in FF Daqu. In the laboratory level simulation of baijiu brewing, the yield of baijiu was increased by 3.36% using FF Daqu.

5.
Foods ; 13(6)2024 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-38540904

RESUMO

Nongxiangxing-daqu (NXDQ), as a saccharification and fermentation agent, directly affects the flavor and yield of fresh Nongxiangxing Baijiu (NXBJ). The difference in fermentation temperature owing to the artificial turning operation leads to the formation of superior (S) and normal (N) grades of NXDQ. Here, aiming to explore the discriminant characteristics of two grades of NXDQ, we studied the physicochemical properties, volatile compounds and microbial communities using HS-SPME-GC/MS and high-throughput sequencing technology. The NXDQ grades presented different physicochemical properties. Staphylococcus, Weissella, Lactobacillus and Thermoascus were dominant in the S grade (S-NXDQ), while Bacillus, Thermoactinomyces and Aspergillus were predominant in the N grade (N-NXDQ). Higher alcohols, aldehydes and ketones positively correlated with the bacterial biomarkers could be used as metabolic biomarkers for N-NXDQ; the S-NXDQ had a higher abundance of key enzymes involved in lactic acid and ethanol fermentation, while N-NXDQ had a higher abundance of key enzymes involved in amino acid synthesis and long-chain fatty acid and lipid metabolism. N-NXDQ and S-NXDQ had different microbial and metabolic biomarkers. These findings provide insight into the discriminant characteristics of different grades of NXDQ, a theoretical basis for rational evaluation of NXDQ, and effective information for quality improvement of daqu.

6.
PeerJ ; 12: e16621, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38188181

RESUMO

Background: Daqu is an essential starter for baijiu brewing in China. However, the microbial enrichment and metabolic characteristics of Daqu formed at different fermentation temperatures are still unclear. Methods: High-throughput sequencing technology and the non-targeted metabolomics were used to compare the microbial communities and metabolites of Taorong-type high-temperature Daqu and middle-temperature Daqu. In this study, the relationship between microorganisms and metabolites was established. Results: The study found that the composition and metabolites of the microbial community differed due to the difference in Daqu-making temperature. The bacterial diversity of Taorong-type high-temperature Daqu was higher than that of middle-temperature Daqu, while the fungal community diversity of Taorong-type middle-temperature Daqu was higher than that of high temperature Daqu. A total of 1,034 differential metabolites were screened from the two types of Daqu, and 76 metabolites with significant differences were detected (P < 0.001 and variable importance in projection (VIP) > 1.15). Tetraacetylethylenediamine is the metabolite with the largest differential fold among the 76 differential metabolites, which can be used as a potential marker metabolite of high-temperature Daqu. Conclusion: This study helps elucidate the microbial assembly mechanisms and functional expression under different processing conditions through a further understanding of the composition and metabolic profile differences of different types of Daqu microflora in Taorong-type baijiu.


Assuntos
Microbiota , Micobioma , Temperatura , Metabolômica , China
7.
J Sci Food Agric ; 103(14): 6849-6860, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37293782

RESUMO

BACKGROUND: There are few reports on the breeding of high-yielding tetramethylpyrazine (TTMP) strains in strong-flavor Daqu. In addition, studies on the mechanism of TTMP production in strains are mostly based on common physiological and biochemical indicators, and there is no report on RNA level. Therefore, in this study, a strain with high production of TTMP was screened out from strong-flavor liquor, and transcriptome sequencing analysis was performed to analyze its key metabolic pathways and key genes, and to infer the mechanism of TTMP production in the strain. RESULTS: In this study, a strain with a high yield of tetramethylpyrazine (TTMP) was screened out, and the yield was 29.83 µg mL-1 . The identified strain was Bacillus velezensis, which could increase the content of TTMP in liquor by about 88%. After transcriptome sequencing, a total of 1851 differentially expressed genes were screened, including 1055 up-regulated genes and 796 down-regulated genes. Three pathways related to the production of TTMP were identified by gene ontology (GO) annotation and COG annotation, including carbohydrate metabolism, cell movement and amino acid metabolism. The key genes of TTMP were analyzed, and the factors that might regulate the production of TTMP, such as the transfer of uracil phosphate ribose and glycosyltransferase, were obtained. CONCLUSIONS: A strain of B. velezensis with high TTMP production was screened and identified in strong-flavor Daqu for the first time. The yield of TTMP was 29.83 µg mL-1 , which increased the TTMP content in liquor by 88%. The key metabolic pathways of TTMP production in the strain were obtained: carbohydrate metabolism, cell movement and amino acid metabolism, and the key regulatory genes of each pathway were found, which complemented the gap in gene level in the production regulation of the strain, and provided a theoretical basis for the subsequent study of TTMP in liquor. © 2023 Society of Chemical Industry.


Assuntos
Metabolismo dos Carboidratos , Pirazinas , Fermentação , Pirazinas/metabolismo , Aminoácidos/metabolismo
8.
Front Microbiol ; 14: 1119675, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36778882

RESUMO

Daqu is a saccharifying and fermenting starter in the production of Chinese Baijiu; its quality directly affects the quality of Baijiu. The production of Daqu is highly environment-dependent, and after long-term natural domestication, it is rich in a wide variety of microorganisms with a stable composition, which provide complex and diverse enzymes and flavor (precursor) substances and microbiota for Jiupei (Fermented grains) fermentation. However, inoculation with a relatively stable microbial community can lead to a certain upper limit or deficiencies of the physicochemical properties (e.g., saccharification capacity, esterification capacity) of the Daqu and affect the functional expression and aroma formation of the Daqu. Targeted improvement of this problem can be proposed by selecting functional microorganisms to fortify the production of Daqu. This review introduced the isolation, screening, identification and functional characteristics of culture-dependent functional microorganisms in Baijiu-brewing, the core functional microbiota community of Daqu, and the related research progress of functional microorganisms fortified Daqu, and summarized the fortifying strategies of functional microorganisms, aiming to further deepen the application of functional microorganisms fortification in Daqu fermentation and provide ideas for the flavor regulation and quality control of Baijiu.

9.
RSC Adv ; 12(55): 35616-35626, 2022 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-36545074

RESUMO

Fruit wine has certain health care functions, but fruit wine made from a single fruit or vegetable does not have a good enough color, flavor or nutrient composition. Therefore, this study used fresh carrot (Daucus carota subsp. sativus) and pomegranate (Punica granatum) as raw materials to explore the brewing process of carrot and pomegranate compound wine. The fermentation technology of the composite carrot and pomegranate wine was optimized by a single-factor experiment and Box-Behnken design (BBD), which provided a theoretical foundation for the fermentation of this wine. As per the results, the alcohol content of this composite carrot and pomegranate wine was 12.35% vol. under the optimum fermentation conditions of 28 °C initial temperature, 24% initial sugar content, and with the addition of 64 mg L-1 sulfur dioxide (SO2). In the fermented fruit and vegetable wine, a total of 30 aroma components were detected; 21 composites (such as bornyl acetate, caryophyllene and 3-(2-nitrophenylmethyl)-2-thiazolidinone) were newly generated. The relative content of alcohol flavor composites (such as propionic acid 2-methyl-3-hydroxy-2,2,4-trimethylpentan-1-ol, 2-methyl-2-ethyl-3-hydroxycyclohexyl propanoate and terpinene-4-ol) showed an upward trend, and the relative content of alkene components increased significantly after fermentation. The findings of this study provide an experimental foundation for optimizing fermentation technology and for improving the product quality of composite carrot and pomegranate wine.

10.
PLoS One ; 17(10): e0274881, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36194604

RESUMO

To study the difference between the fungal community compositional and fragrance components in medium- and high-Temperature Taorong-type Baijiu Daqu. The microbial communities and fragrance components of Taorong-type Baijiu Daqu were analyzed using high-throughput sequencing (HTS) and headspace-solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). With an abundance at the phylum and genus levels ≥0.01% as the threshold, 3 phyla, Mucoromycota, Ascomycota, and Basidiomycota, were found in both medium- and high-temperature Daqu, but their abundances differed. At the genus level, 15 and 13 genera were recognized. Rhizopus (72.40%) and Thermomyces (53.32%) accounted for the most significant proportions in medium-temperature and high-temperature Daqu, respectively. Medium-temperature Daqu and high-temperature Daqu were found to have 40 and 29 fragrance components, respectively and contained the highest proportions of pyrazines (53.12%) and acids (32.68%). Correlation analyses between microbes and fragrance components showed that Aspergillus, Hyphopichia, Trichosporon, Alternaria were all highly and positively correlated with pyrazines, but the dominant fungal communities were highly correlated with only a few individual acid compounds but not with acid compounds overall. The unique Daqu -making process and environment lead to these differences.


Assuntos
Micobioma , Bebidas Alcoólicas/análise , Bactérias , Fermentação , Pirazinas , Temperatura
11.
Microb Cell Fact ; 21(1): 194, 2022 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-36123739

RESUMO

BACKGROUND: Due to the detrimental effects of chemical preservatives, there has been an increasing demand for safer, healthier and natural bio-preservatives. Bacteriocins have attracted increasing interest because of their potential as natural bio-preservatives. RESULTS: We screened a large number of Bacillus thuringiensis strains and isolated one strain (B. thuringiensis P86) with antimicrobial activity against several foodborne pathogens. Three novel leaderless bacteriocins, including thucin A1, thucin A2 and thucin A3, were purified and identified from the culture supernatant of B. thuringiensis P86, whose molecular masses were 5552.02, 5578.07 and 5609.06 Da, respectively. Thucin A1 was then selected as a representative to be tested, and it exhibited potent inhibitory activity against all tested gram-positive bacteria. More importantly, thucin A1 showed stronger antimicrobial activity than nisin A against two important foodborne pathogens Bacillus cereus and Listeria monocytogenes. In addition, thucin A1 exhibited strong acid-base adaptability (pH 2-11), high endurance to heat, good stability to trypsin and pepsin, no hemolysis activity and cytotoxicity, and could effectively inhibit or eliminate Bacillus cereus and Listeria monocytogenes in skim milk. CONCLUSIONS: Our findings indicate that these novel leaderless bacteriocins are potentially promising food biopreservatives.


Assuntos
Anti-Infecciosos , Bacteriocinas , Listeria monocytogenes , Anti-Infecciosos/farmacologia , Bacillus cereus , Bacteriocinas/química , Bactérias Gram-Positivas , Testes de Sensibilidade Microbiana , Pepsina A/farmacologia , Tripsina
12.
Front Microbiol ; 13: 934696, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35966713

RESUMO

Medium-temperature Daqu (MT-Daqu), a saccharification-fermentation agent and aroma-producing agent, is used to produce Chinese strong-flavor Baijiu. Many related studies have been published; however, less is known about microbial community and quality properties inside and outside the MT-Daqu from fermentation to storage. Here, along with determining the physicochemical index, the microbial community of MT-Daqu was investigated using both culture-dependent and culture-independent methods during 31 days of fermentation and 4 months of storage. Volatile compounds of mature MT-Daqu were analyzed using headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS). The results indicated obvious variation in the microbial community due to the changes in environmental conditions, and the physicochemical indices shifted from fluctuations in the fermentation period to relative stability after storage for 3 months. Moreover, the microbial counts and physicochemical indices of the inner layers of MT-Daqu differed from those of the outer layers. The dominant communities, including the bacterial phyla Firmicutes, Proteobacteria, and Actinobacteria and the fungal phyla Ascomycota and Mucoromycota, showed different abundances in the two parts of the mature MT-Daqu, and different microbial communities were enriched in both parts. Additionally, pyrazines and alcohols were the most abundant volatile aroma compounds in the mature MT-Daqu.

13.
Food Sci Nutr ; 10(6): 1780-1788, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35702309

RESUMO

Currently, the primary method of identifying high- and low-quality liquors is sensory tasting, which is prone to uncertainty caused by the biases of tasters. To address this problem, this study used color, aroma, taste, and style as four factors affecting the sensory quality of Luzhou-flavor liquor; determined the weights of each factor; and quantitatively evaluated the sensory quality of five different Luzhou flavor liquor using fuzzy mathematical methods. The volatile aromatic substances in the liquor samples were detected by GC-MS, and analyzed using principal component analysis. The results obtained from fuzzy mathematics and principal component analysis indicated that the comprehensive evaluation system was scientifically sound and reasonably constructed.

14.
Sci Rep ; 12(1): 7347, 2022 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-35513386

RESUMO

In this study, the pit mud used in manufacturing Taorong-type Baijiu was collected from the upper, middle, lower and bottom layers of pits at Henan Yangshao Liquor Co., LTD. High-throughput sequencing (HTS) technology was used to analyze the microbial community structure of the pit mud. In addition, the volatile compounds in the pit mud were subjected to preliminary qualitative analysis through headspace-solid phase microextraction and gas chromatography-mass spectrometry (GC-MS). The HTS results demonstrated that there were 5, 3, 5 and 5 dominant bacterial phyla (including 11, 11, 9 and 8 dominant bacterial genera) and 3, 3, 3 and 3 dominant fungal phyla (including 4, 7, 7 and 5 dominant fungal genera) in the pit mud from the F-S (upper), G-Z (middle), H-X (lower) and I-D (bottom) layers, respectively. In the qualitative analysis of the volatile compounds, a total of 77types of volatile compounds were detected in the pit mud, including 46, 45, 39 and 49 types in the pit mud from layers F-S, G-Z, H-X and I-D, respectively. Esters and acids were the two main components of the pit mud. The correlation between the microorganisms present and the main volatile compounds in the pit mud was analyzed. Lentimicrobium, Syner-01 and Blvii28_wastewater-sludge groups were found for the first time in pit mud used for manufacturing Taorong-type Baijiu. The findings of this study could provide a theoretical foundation for improving the quality of pit mud and the flavor of Taorong-type Baijiu.


Assuntos
Microbiota , Compostos Orgânicos Voláteis , Bebidas Alcoólicas/análise , Bactérias/genética , Fermentação , Sequenciamento de Nucleotídeos em Larga Escala , Microbiota/genética , Compostos Orgânicos Voláteis/análise
15.
PLoS One ; 17(4): e0264677, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35472204

RESUMO

A total of 296 strains of protease-producing bacteria were isolated and purified from medium-temperature Daqu produced by ZhangGong LaoJiu Wine Co. Ltd. After calculating the ratio of transparent ring diameter to colony diameter and measuring the protease activities, a strain of high-yield protease bacteria, called DW-7, was screened out with a protease activity of 99.54 U/mL. Through morphological observation, 16S rDNA sequence analysis, and physiological and biochemical tests, the isolated bacteria DW-7 was determined to be Bacillus velezensis. In addition, whole-genome sequencing (WGS), using PacBio and the Illumina platform, was performed. Gene annotation was then conducted using the Clusters of Orthologous Groups (COG), Kyoto Encyclopedia of Genes and Genomes (KEGG), Non-Redundant Protein Sequence Database (NR), and Gene Ontology (GO) databases. The results showed that the genome of DW-7 was 3,942,829 bp long with a GC content of 46.45%. A total of 3,662 protein-encoding genes were predicted, with a total length of 3,402,822 bp. Additionally, 2,283; 2,796; and 2,127 genes were annotated in the COG, KEGG, and GO databases, respectively. A total of 196 high-yield protease genes were mainly enriched in the metabolism of alanine, aspartic acid, glutamate, glycine, serine, and threonine, as well as ABC transporter and transporter pathways.


Assuntos
Peptídeo Hidrolases , Transcriptoma , Bactérias/genética , Perfilação da Expressão Gênica/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Anotação de Sequência Molecular , Peptídeo Hidrolases/genética
16.
Biochem Biophys Res Commun ; 571: 32-37, 2021 09 24.
Artigo em Inglês | MEDLINE | ID: mdl-34303193

RESUMO

Flowering time or heading date is a critical agronomic trait of rice and is regulated by numerous genes, including several genes encoding nuclear factor YB (NF-YB) in rice, NF-YB11 is one of the genes well known to be involved in the process, delaying flowering under long-day (LD) conditions but promoting flowering under short-day (SD) conditions. In this study, we identified another NF-YB gene, OsNF-YB4. Overexpression of OsNF-YB4 promoted rice flowering under both natural long-day (NLD) and natural short-day (NSD) conditions, whereas suppression or loss-of-function of this gene delayed flowering. The transcription of OsNF-YB4 exhibited an obvious circadian pattern and was induced by light under both LD and SD conditions. Expression analyses of flowering regulators in the photoperiodic flowering pathway demonstrated that up-regulation of OsNF-YB4 resulted in down-regulation of floral repressor Grain number, plant height and heading date 7 (Ghd7), and thus activating the Early heading date 1 (Ehd1)-mediated flowering pathway. Besides, OsNF-YB4 was observed to bind to the specific CCAAT-box regions in the Ghd7 promoter in vitro and interact with GHD7 in yeast. All these evidences support that OsNF-YB4 functions as a flowering promoter by negatively regulating the expression of floral repressor Ghd7 in rice photoperiodic flowering-time regulatory network.


Assuntos
Flores/metabolismo , Proteínas Nucleares/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Flores/genética , Regulação da Expressão Gênica de Plantas/genética , Proteínas Nucleares/genética , Oryza/genética , Proteínas de Plantas/genética
17.
Biotechnol Biofuels ; 14(1): 11, 2021 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-33413621

RESUMO

BACKGROUND: Efficient utilization of all available carbons from lignocellulosic biomass is critical for economic efficiency of a bioconversion process to produce renewable bioproducts. However, the metabolic responses that enable Pseudomonas putida to utilize mixed carbon sources to generate reducing power and polyhydroxyalkanoate (PHA) remain unclear. Previous research has mainly focused on different fermentation strategies, including the sequential feeding of xylose as the growth stage substrate and octanoic acid as the PHA-producing substrate, feeding glycerol as the sole carbon substrate, and co-feeding of lignin and glucose. This study developed a new strategy-co-feeding glycerol and lignin derivatives such as benzoate, vanillin, and vanillic acid in Pseudomonas putida KT2440-for the first time, which simultaneously improved both cell biomass and PHA production. RESULTS: Co-feeding lignin derivatives (i.e. benzoate, vanillin, and vanillic acid) and glycerol to P. putida KT2440 was shown for the first time to simultaneously increase cell dry weight (CDW) by 9.4-16.1% and PHA content by 29.0-63.2%, respectively, compared with feeding glycerol alone. GC-MS results revealed that the addition of lignin derivatives to glycerol decreased the distribution of long-chain monomers (C10 and C12) by 0.4-4.4% and increased the distribution of short-chain monomers (C6 and C8) by 0.8-3.5%. The 1H-13C HMBC, 1H-13C HSQC, and 1H-1H COSY NMR analysis confirmed that the PHA monomers (C6-C14) were produced when glycerol was fed to the bacteria alone or together with lignin derivatives. Moreover, investigation of the glycerol/benzoate/nitrogen ratios showed that benzoate acted as an independent factor in PHA synthesis. Furthermore, 1H, 13C and 31P NMR metabolite analysis and mass spectrometry-based quantitative proteomics measurements suggested that the addition of benzoate stimulated oxidative-stress responses, enhanced glycerol consumption, and altered the intracellular NAD+/NADH and NADPH/NADP+ ratios by up-regulating the proteins involved in energy generation and storage processes, including the Entner-Doudoroff (ED) pathway, the reductive TCA route, trehalose degradation, fatty acid ß-oxidation, and PHA biosynthesis. CONCLUSIONS: This work demonstrated an effective co-carbon feeding strategy to improve PHA content/yield and convert lignin derivatives into value-added products in P. putida KT2440. Co-feeding lignin break-down products with other carbon sources, such as glycerol, has been demonstrated as an efficient way to utilize biomass to increase PHA production in P. putida KT2440. Moreover, the involvement of aromatic degradation favours further lignin utilization, and the combination of proteomics and metabolomics with NMR sheds light on the metabolic and regulatory mechanisms for cellular redox balance and potential genetic targets for a higher biomass carbon conversion efficiency.

18.
Bioresour Technol ; 273: 538-544, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30472353

RESUMO

Nitrogen supply is critical to the synthesis of intracellular PHA in various bacteria. However, the specific role of the nitrogen in synthesizing PHA from benzoate, a lignin model compound use for the study of bacteria catabolism of aromatics, is still not clear. In this study, two culture conditions were maintained for Pseudomonas putida KT2440 to produce PHA using benzoate as a carbon source. Under nitrogen-limited and surplus conditions, the accumulation of PHA was to 37.3% and 0.25% of cell dry weight, respectively. A model fit to the kinetics of biomass growth and PHA accumulation showed good agreement with data. GC-MS and NMR showed that PHA contained six hydroxyl fatty acid monomers under nitrogen-limited conditions, while two monomers were identified under nitrogen surplus conditions. The average molecular weight of PHA increased after the nitrogen source was exhausted. These results provide a promising strategy for optimization of lignin to PHA yields.


Assuntos
Benzoatos/metabolismo , Nitrogênio/metabolismo , Poli-Hidroxialcanoatos/biossíntese , Pseudomonas putida/metabolismo , Biomassa , Carbono/metabolismo , Ácidos Graxos/metabolismo , Cinética
19.
Curr Microbiol ; 74(12): 1389-1393, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28840359

RESUMO

The Gram-stain-negative, rod-shaped and non-motile bacterial strain, designated ZGLJ7-1T, was isolated from a pit mud. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain ZGLJ7-1T was related to the genus Lysobacter and had the highest 16S rRNA gene sequence similarity with the type strain of Lysobacter arseniciresistens ZS79T (97.4%). The predominant cellular fatty acids were iso-C15:0, iso-C17:1ω9c, iso-C11:0 and iso-C11:03-OH. Strain ZGLJ7-1T had Q-8 as the predominant ubiquinone. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, one unidentified phospholipid, two unidentified aminolipids and two unidentified lipids. The genomic DNA G+C content of strain ZGLJ7-1T was 69.5 mol%. Strain ZGLJ7-1T shared DNA relatedness with 35% Lysobacter arseniciresistens CGMCC 1.10752T. Combined data from phenotypic, phylogenetic and DNA-DNA relatedness studies demonstrated that the strain ZGLJ7-1T is a representative of a novel species of the genus Lysobacter, for which we propose the name Lysobacter zhanggongensis sp. nov. (type strain ZGLJ7-1T = KACC 18547T = CGMCC 1.15404T).


Assuntos
Lysobacter/classificação , Lysobacter/isolamento & purificação , Microbiologia do Solo , Composição de Bases , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Locomoção , Lysobacter/genética , Lysobacter/fisiologia , Microscopia Eletrônica de Varredura , Hibridização de Ácido Nucleico , Fosfolipídeos/análise , Filogenia , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
20.
Environ Sci Pollut Res Int ; 22(23): 18394-403, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26206124

RESUMO

The enhanced H2 production from maize straw had been achieved through the two-stage process of integrating H2 fermentation and microbial electrolysis cells (MECs) in the present work. Several key parameters affecting hydrolysis of maize straw through subcritical H2O were optimized by orthogonal design for saccharification of maize straw followed by H2 production through H2 fermentation. The maximum reducing sugar (RS) content of maize straw reached 469.7 mg/g-TS under the optimal hydrolysis condition with subcritical H2O combining with dilute HCl of 0.3% at 230 °C. The maximum H2 yield, H2 production rate, and H2 content was 115.1 mL/g-TVS, 2.6 mL/g-TVS/h, and 48.9% by H2 fermentation, respectively. In addition, the effluent from H2 fermentation was used as feedstock of MECs for additional H2 production. The maximum H2 yield of 1060 mL/g-COD appeared at an applied voltage of 0.8 V, and total COD removal reached about 35%. The overall H2 yield from maize straw reached 318.5 mL/g-TVS through two-stage processes. The structural characterization of maize straw was also carefully investigated by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) spectra.


Assuntos
Bactérias/metabolismo , Fontes de Energia Bioelétrica/microbiologia , Hidrogênio/metabolismo , Caules de Planta/microbiologia , Zea mays/microbiologia , Eletrólise , Fermentação , Hidrogênio/química , Hidrólise , Microbiologia Industrial , Caules de Planta/química , Caules de Planta/metabolismo , Resíduos/análise , Difração de Raios X , Zea mays/química , Zea mays/metabolismo
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