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1.
Int J Biol Macromol ; 247: 125628, 2023 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-37392926

RESUMO

A strain of Bacillus that can tolerate 10 g/L acetic acid and use the volatile fatty acids produced by the hydrolysis and acidification of activated sludge to produce polyhydroxyalkanoate was screened from the activated sludge of propylene oxide saponification wastewater. The strain was identified by 16S rRNA sequencing and phylogenetic tree analysis and was named Bacillus cereus L17. Various characterization methods showed that the polymer synthesized by strain L17 is poly-ß-hydroxybutyrate, which has low crystallinity, good ductility and toughness, high thermal stability and a low polydispersity coefficient. It has wide thermoplastic material operating space as well as industrial and medicinal applications. The optimal fermentation conditions were determined by single factor optimization. Then, Plackett-Burman and Box-Behnken design experiments were carried out according to the single factor optimization results, and the response surface optimization was completed. The final results were: initial pH 6.7, temperature 25 °C, and loading volume 124 mL. The verification experiment showed that the yield of poly-ß-hydroxybutyrate after optimization increased by 35.2 % compared to that before optimization.


Assuntos
Bacillus cereus , Esgotos , Bacillus cereus/metabolismo , Ácido Acético , Carbono , RNA Ribossômico 16S/genética , Filogenia , Poliésteres/química , Fermentação , Hidroxibutiratos/química
2.
Int J Biol Macromol ; 234: 123667, 2023 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-36796552

RESUMO

The microflora in the activated sludge of propylene oxide saponification wastewater is characterized by a clear succession after enrichment and domestication, and the specifically enriched strains can significantly increase the yield of polyhydroxyalkanoate. In this study, Pseudomonas balearica R90 and Brevundimonas diminuta R79, which are dominant strain after domestication, were selected as models to examine the interactive mechanisms associated with the synthesis of polyhydroxyalkanoate by co-cultured strains. RNA-Seq analysis revealed the up-regulated expression of the acs and phaA genes of strains R79 and R90 in the co-culture group, which enhanced their utilization of acetic acid and synthesis of poly-ß-hydroxybutyrate. Cell dry weight and the yield of poly-ß-hydroxybutyrate in the co-culture group were accordingly considerably higher than those in the respective pure culture groups. In addition, two-component system, quorum-sensing, flagellar synthesis-related, and chemotaxis-related genes were enriched in strain R90, thereby indicating that compared with the R79 strain, R90 can adapt more rapidly to a domesticated environment. Expression of the acs gene was higher in R79 than in R90, and consequently, strain R79 could more efficiently assimilate acetate in the domesticated environment, and thus predominated in the culture population at the end of the fermentation period.


Assuntos
Poli-Hidroxialcanoatos , Pseudomonas/genética , Pseudomonas/metabolismo , Poliésteres/metabolismo , Hidroxibutiratos
3.
Bioresour Technol ; 348: 126786, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35114368

RESUMO

In this study, novel polyhydroxyalkanoate (PHA)-associated genes (phaCp and phaABp) cloned from Propylenella binzhouense L72T were expressed in Escherichiacoli cells for PHA production, and the recombinant strains were used to analyze PHA yields with various substrates. The highest poly (3-hydroxybutyrate-co-3-hydroxy-valerate) (PHBV) yield (1.06 g/L) and cell dry weight (3.31 g/L) in E. coli DH5α/ΔptsG-CpABp were achieved by using glucose and propionicacid as substrates. Structural verification of PHBV produced by E. coli DH5α/ΔptsG-CpABp was performed to evaluate the characteristics of the polymers using Fourier transform infrared spectroscopy and nuclear magnetic resonance analysis. In addition, the X-ray diffraction results showed improved crystallinity of PHBV, and thermogravimetric analysis showed good thermal stability of 298 °C. The above findings indicated that the expression of phaCp and phaABp genes resulted in improved PHBV synthesis activity, and the polymer had better performance at higher processing temperatures.


Assuntos
Engenharia Metabólica , Poli-Hidroxialcanoatos , Escherichia coli/genética , Escherichia coli/metabolismo , Glucose/metabolismo , Engenharia Metabólica/métodos , Poliésteres/metabolismo , Propionatos
4.
J Biomed Nanotechnol ; 17(9): 1726-1734, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34688317

RESUMO

The aim of this study was to test an effective nano-pole capsule loaded cis-platinum (CP) transplantation device for liver cancer (LC) therapy. A novel nano-pole capsule was designed as a new vector for storing CP. HepG2 cells and a B6/J mouse model were used to test the efficiency of polyethyleneimine-cis-platinum (PEI-CP) and poly-chitosan-cis-platinum (PC-CP). Infiltration efficiency and transplantation efficiency tests were performed to study the performance of the delivery system, and fibroblast reactions and macrophage numbers were observed, to test for immune rejection and foreign body reactions. The apoptosis rate and tumor diameter of hepatocellular carcinoma cells were used to evaluate the effect of the tumor therapy. We also studied the functional mechanism of different CP delivery systems. The infiltration and transplantation efficiencies of PC-CP were higher than that of PEI-CP; Less foreign body reaction appeared in PC system, with less fibroblast reaction and lower macrophage reaction. The clinical efficacy of PC-CP in terms of tumor apoptosis and diameter reduction was superior to that of PEI-CP. We demonstrated that PC-CP had a more significant alteration effect on mTOR, P-Ak, LC3 and P53. The PC system can better deliver and release drugs than PEI-CP, and may be a better choice for LC therapy in the future.


Assuntos
Quitosana , Neoplasias Hepáticas , Nanopartículas , Preparações Farmacêuticas , Animais , Cisplatino , Neoplasias Hepáticas/tratamento farmacológico , Camundongos , Polietilenoimina , Transfecção
5.
Int J Biol Macromol ; 193(Pt A): 956-964, 2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34751142

RESUMO

In this study, a phaCR gene encoding PHA synthase was identified in Rhodoligotrophos defluvii which was adjacent to ß-ketothiolase encoded by phaAR gene and acetoacetyl-CoA reductase encoded by phaBR gene. Amino acid comparison of PhaCR showed the highest homology of 65.98% with PhaC of R. appendicifer, while its homology with typical class I PHA synthase in Cupriavidus necator was only 42.54%. PHA synthesis genes were then transformed into E. coli harboring phaCABR and phaCRABC which were cultured with 15 g/L glucose respectively, and 20.46 wt% and 16.95 wt% of CDW for poly(3-hydroxybutyrate) (PHB) were accumulated respectively. To further explore the effect of substrate specificity for PHA production, the ptsG gene was then deleted and 15 g/L glucose and 1.5 g/L propionate were co-employed as carbon sources, which enabled the synthesis of poly(3HB-co-3HV) copolymer. As a result, poly(3HB-co-3HV) was accumulated up to 24.74 wt% of CDW, and the highest content of 3-hydroxyvalerate (3HV) was 10.86 mol%. The Td5 was 260 °C, which implied that it possessed good thermal stability, and the Mw of GPC in recombinant strains were between 22 and 26 × 104 g/mol, and the highest PDI was 3.771. The structure of poly (3HB-co-3HV) copolymer was determined through 1H NMR analysis.


Assuntos
Escherichia coli/genética , Engenharia Metabólica/métodos , Poliésteres/metabolismo
6.
PeerJ ; 5: e4057, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29158986

RESUMO

Prophages have been considered genetic units that have an intimate association with novel phenotypic properties of bacterial hosts, such as pathogenicity and genomic variation. Little is known about the genetic information of prophages in the genome of Streptococcus mutans, a major pathogen of human dental caries. In this study, we identified 35 prophage-like elements in S. mutans genomes and performed a comparative genomic analysis. Comparative genomic and phylogenetic analyses of prophage sequences revealed that the prophages could be classified into three main large clusters: Cluster A, Cluster B, and Cluster C. The S. mutans prophages in each cluster were compared. The genomic sequences of phismuN66-1, phismuNLML9-1, and phismu24-1 all shared similarities with the previously reported S. mutans phages M102, M102AD, and ϕAPCM01. The genomes were organized into seven major gene clusters according to the putative functions of the predicted open reading frames: packaging and structural modules, integrase, host lysis modules, DNA replication/recombination modules, transcriptional regulatory modules, other protein modules, and hypothetical protein modules. Moreover, an integrase gene was only identified in phismuNLML9-1 prophages.

7.
Int J Biol Macromol ; 98: 34-38, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28130132

RESUMO

The saponification wastewater from the process of propylene oxide (PO) production is contaminated with high chemical oxygen demand (COD) and chlorine contents. Although the activated sludge process could treat the PO saponification wastewater effectively, the residual sludge was difficult to be disposed properly. In this research, microbes in PO saponification wastewater residual sludge were acclimated to produce poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) from volatile fatty acids. Through Miseq Illumina highthroughput sequencing, the bacterial community discrepancy between the original and the acclimated sludge samples were analyzed. The proportions of Bacillus, Acinetobacter, Brevundimonas and Pseudomonas, the potential PHBV-producers in the residual sludge, were all obviously increased. In the batch fermentation, the production of PHBV could achieve 4.262g/L at 300min, with the content increased from 0.04% to 23.67% of mixed liquor suspended solid (MLSS) in the acclimated sludge, and the COD of the PO saponification wastewater was also decreased in the fermentation. This work would provide an effective solution for the utilization of PO saponification wastewater residual sludge.


Assuntos
Bactérias/metabolismo , Compostos de Epóxi/metabolismo , Poliésteres/metabolismo , Esgotos/microbiologia , Acetatos/metabolismo , Análise da Demanda Biológica de Oxigênio , Reatores Biológicos/microbiologia , Fermentação
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