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Bioengineered ; 10(1): 335-344, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31322471


Selenium-enriched yeast can transform toxic inorganic selenium into absorbable organic selenium, which is of great significance for human health and pharmaceutical industry. A yeast Rhodotorula glutinis X-20 we obtained before has good selenium-enriched ability, but its selenium content is still low for industrial application. In this study, strategies of process optimization and transport regulation of selenium were thus employed to further improve the cell growth and selenium enrichment. Through engineering phosphate transporters from Saccharomyces cerevisiae into R. glutinis X-20, the selenium content was increased by 21.1%. Through using mixed carbon culture (20 g L-1, glycerol: glucose 3:7), both biomass and selenium content were finally increased to 5.3 g L-1 and 5349.6 µg g-1 (cell dry weight, DWC), which were 1.14 folds and 6.77 folds compared to their original values, respectively. Our results indicate that high selenium-enrichment ability and biomass production can be achieved through combining process optimization and regulation of selenium transport.

Engenharia Metabólica/métodos , Fosfatos/metabolismo , Rhodotorula/genética , Saccharomyces cerevisiae/genética , Selênio/metabolismo , Transgenes , Transporte Biológico , Biomassa , Meios de Cultura/química , Meios de Cultura/farmacologia , Fermentação , Expressão Gênica , Glucose/química , Glucose/metabolismo , Glicerol/química , Glicerol/metabolismo , Proteínas de Transporte de Fosfato/genética , Proteínas de Transporte de Fosfato/metabolismo , Plasmídeos/química , Plasmídeos/metabolismo , Simportadores de Próton-Fosfato/genética , Simportadores de Próton-Fosfato/metabolismo , Rhodotorula/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas Cotransportadoras de Sódio-Fosfato Tipo III/genética , Proteínas Cotransportadoras de Sódio-Fosfato Tipo III/metabolismo
Biosens Bioelectron ; 127: 207-214, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30611108


This work describes a hybrid electrochemical sensor for highly sensitive detection of pesticide cypermethrin (CYP). Firstly, Ag and N co-doped zinc oxide (Ag-N@ZnO) was produced by sol-gel method, and then Ag-N@ZnO was ultrasonically supported on activated carbon prepared from coconut husk (Ag-N@ZnO/CHAC). Finally, a layer of molecularly imprinted polymer (MIP) was in situ fabricated on glassy carbon electrode by electro-polymerization, with dopamine and resorcinol as dual functional monomers (DM), CYP acting as template (DM-MIP-Ag-N@ZnO/CHAC). Morphological features, composition information and electrochemical properties of DM-MIP-Ag-N@ZnO/CHAC were investigated in detail. It is worth to mention that for the first time response surface method was used to investigate the effect of double monomers and to optimize the ratio between template and monomers. Compared with typical one-monomer involving MIP, the MIP prepared with dual functional monomers (DMMIP) of monomers showed higher response and better selectivity. Under the optimal conditions, a calibration curve of current shift versus concentration of CYP was obtained in the range of 2 × 10-13~8 × 10-9 M, and the developed sensor gave a remarkably low detection limit (LOD) of 6.7 × 10-14 M (S/N = 3). Determination of CYP in real samples was conducted quickly and accurately with our sensor. The DMMIP-Ag-N@ZnO/CHAC electrochemical sensor proposed in this paper has great potential in food safety, drug residue determination and environmental monitoring.

Técnicas Biossensoriais , Técnicas Eletroquímicas , Polímeros/química , Piretrinas/isolamento & purificação , Carbono/química , Eletrodos , Grafite/química , Limite de Detecção , Impressão Molecular , Polimerização , Piretrinas/química
Food Funct ; 10(1): 266-276, 2019 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-30564811


Enzyme-assisted aqueous two-phase extraction (EA-ATPE) using ethanol/ammonium sulfate system was investigated for total polyphenol (TP) and lutein from marigold flowers. The important factors were investigated by single factor experiment and response surface methodology combined with Box-Behnken design to optimize the operating parameters of EA-ATPE. The maximum yields of TP and lutein were 83.56 ± 0.69 mg g-1 and 5.59 ± 0.13 mg g-1, respectively. Compared with aqueous two-phase extraction and Soxhlet extraction (SE), the extraction yield of TP by EA-ATPE is 64.91% higher and the extract of EA-ATPE has better antioxidant activity. The pretreatment effect was also researched by scanning electron microscopy. Thus, EA-ATPE is an efficient method for extracting bioactive components from plants.

Luteína/isolamento & purificação , Extratos Vegetais/isolamento & purificação , Polifenóis/isolamento & purificação , Tagetes/química , Sulfato de Amônio/química , Fracionamento Químico , Etanol/química , Flores/química , Luteína/análise , Extratos Vegetais/análise , Polifenóis/análise
Biosens Bioelectron ; 106: 71-77, 2018 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-29414092


A highly sensitive electrochemical sensor using a carbon paste electrode (CPE) modified with surface molecularly imprinted polymeric microspheres (SMIPMs) was developed for methyl parathion (MP) detection. Molecular imprinting technique based on distillation precipitation polymerization was applied to prepare SMIPMs and non-surface imprinted microspheres (MIPMs). The polymer properties including morphology, size distribution, BET specific surface area and adsorption performance were investigated and compared carefully. Both MIPMs and SMIPMs were adopted to prepare CPE sensors and their electrochemical behaviors were characterized via cyclic voltammetry and electrochemical impedance spectroscopy. Compared with MIPMs packed sensor, SMIPMs/CPE exhibits a higher sensing response towards MP with linear detection range of 1 × 10-12-8 × 10-9 mol L-1 and detection limit of 3.4 × 10-13 mol L-1 (S/N = 3). Moreover, SMIPMs/CPE exhibits good selectivity and stability in multiple-cycle usage and after long-time storage. Finally, the developed sensor was used to determine MP in real samples including soil and vegetables and only simple pretreatment is needed. The detection results were consistent with those obtained from liquid chromatography. Collectively, this newly developed sensor system shows significant potential for use in a variety of fields like food safety, drug residue determination and environmental monitoring.

Técnicas Biossensoriais , Técnicas Eletroquímicas , Metil Paration/isolamento & purificação , Carbono/química , Limite de Detecção , Metil Paration/química , Microesferas , Impressão Molecular , Polímeros/química
Bioengineered ; 7(6): 432-438, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27442598


Microbial production of 2,3-butanediol is limited by the toxic components in the lignocellulose hydrolysate. To improve the 2,3-butanediol production via Klebsiella pneumoniae from cotton stalk hydrolysate, a method coupling a high tolerance of strain and detoxification of the hydrolysate was thus investigated in this study. The strain tolerance of K. pneumoniae to the cotton stalk hydrolysate was improved via an adaptive laboratory evolution, which involved a stepwise increase in the hydrolysate concentration in the medium. Compared with the initial strain, the resulting strain increased the biomass 3.2-fold in a medium of 20 g/L hydrolysate and produced 10.45 g/L of 2,3-butanediol at an optimal concentration of 60 g/L hydrolysate. After detoxification of cotton stalk hydrolysate, the cell metabolism of K. pneumoniae was further promoted, and the 2,3-butanediol production increased by 1.2 folds. Using fed-batch fermentation, the concentration of 2,3-butanediol reached 35.5 g/L with a yield of 0.43 g/g. The results demonstrated that the bioconversion of low-cost cotton stalk hydrolysate into 2,3-butanediol improves the economics of microbial 2,3-butanediol production.

Butileno Glicóis/metabolismo , Microbiologia Industrial/métodos , Klebsiella pneumoniae/metabolismo , Técnicas de Cultura Celular por Lotes , Biomassa , Evolução Molecular , Fermentação , Lignina/química