Construction of an l-cysteine hyper-producing strain of Escherichia coli based on a balanced carbon and sulfur module strategy / 生物工程学报

l-cysteine is an important sulfur-containing α-amino acid. It exhibits multiple physiological functions with diverse applications in pharmaceutical cosmetics and food industry. Here, a strategy of coordinated gene expression between carbon and sulfur modules in Escherichia coli was proposed and conducted for the production of l-cysteine. Initially, the titer of l-cysteine was improved to (0.38±0.02) g/L from zero by enhancing the biosynthesis of l-serine module (serAf, serB and serCCg) and overexpression of CysB. Then, promotion of l-cysteine transporter, increased assimilation of sulfur, reduction or deletion of l-cysteine and l-serine degradation pathway and enhanced expression of cysEf (encoding serine acetyltransferase) and cysBSt (encoding transcriptional dual regulator CysB) were achieved, resulting in an improved l-cysteine titer (3.82±0.01) g/L. Subsequently, expressions of cysM, nrdH, cysK and cysIJ genes that were involved in sulfur module were regulated synergistically with carbon module combined with utilization of sulfate and thiosulfate, resulting in a strain producing (4.17±0.07) g/L l-cysteine in flask shake and (11.94±0.1) g/L l-cysteine in 2 L bioreactor. Our results indicated that efficient biosynthesis of l-cysteine could be achieved by a proportional supply of sulfur and carbon in vivo. This study would facilitate the commercial bioproduction of l-cysteine.

Draft genome sequence of sulfur-reducing archaeon Thermococcus thioreducens DSM 14981T

Abstract Thermococcus thioreducens DSM 14981T, a sulfur-reducing archaeon, was isolated from the rainbow hydrothermal vent site on the Mid-Atlantic Ridge. Herein, we report the draft genome sequence of T. thioreducens DSM 14981T; we obtained 41 contigs with a genome size of 2,052,483 bp and G + C content of 53.5%. This genome sequence will not only help understand how the archaeon adapts to the deep-sea hydrothermal environment but also aid the development of enzymes that are highly stable under extreme conditions for industrial applications.

Efeitos de fontes orgânicas de cobre e enxofre sobre os parâmetros bioquímicos no soro de ovinos / Effects of organic and inorganic copper and sulphur supplementation on blood biochemical parameters in sheep

Minerais são componentes essenciais na dieta, exercendo diversas funções no organismo animal onde o uso de sais orgânicos visa aumentar a disponibilidade dos inerais no trato digestório. O perfil metabólico auxilia na avaliação de índices produtivos, para tanto, se faz análises de componentes bioquímicos do sangue. O objetivo foi estudar os efeitos que fontes orgânicas e inorgânicas de cobre e enxofre possuem nos parâmetros bioquímicos. O experimento foi realizado na FZEA/USP, para tanto 40 ovinos foram distribuídos em 10tratamentos: 1) dieta basal; 2) dieta contendo Mo; 3) dieta basal+ Cu inorg + S inorg; 4) dieta basal + Cu inorg + S org; 5) dieta basal + Cu org + S inorg; 6) dieta basal + Cu org + S org; 7) dieta com Mo + Cu inorg + S inorg; 8) dieta com Mo + Cu inorg + S org; 9) dieta com Mo + Cu org + S inorg; 10) dieta com Mo + Cu org + S org. De acordo com cada tratamento houve a inclusão de 10mgkg-1 de MS de Cu inorgânico ou orgânico ou 10 mg kg-1 de MS de Mo ou 0,2% de S inorgânico ou orgânico. Os animais receberam dieta única duas vezes ao dia com inclusão de volumoso num total de 3% do peso vivo. O experimento teve duração de 84 dias, com coletas a cada 28 dias para estudo de glicose, ureia, albumina, colesterol, triglicerídeos. Os parâmetros foram analisados com estrutura fatorial 2 x 2 x 2 (com e sem Mo, Cu orgânico e inorgânico e S orgânico e inorgânico) e uma dieta basal e uma basal mais molibdênio, em delineamento inteiramente casualizado, com um nível de significância de 5%. Os teores séricos de glicose, ureia, albumina e colesterol não apresentaram diferença significativa entre tratamentos e tempo, tão pouco foram influenciados (p>0.05) pelos tratamentos, sendo as médias 64,8mg dl-1; 30,0mg dl-1; 2,78mg L-1; 72,2mg dl-1respectivamente, apresentando-se dentro da normalidade. Os teores de triglicerídeos nos tratamentos (28.8; 34.8; 30.8; 36.9; 34.3; 27.0; 31.6; 32.1; 34.6; 31.1mg dl-1) foram influenciados pela interação Cu x S...

Minerals are essential components in the diet, exercising various functions in the animal. There are many studies to find better results for the organic sources of minerals, and one of the reasons for that is to avoid the negative interactions. The metabolic profile helps in assessment of production indices, therefore analyzes blood biochemical components. The objective of this study was to investigate the effects of organic and inorganic copper and sulphur supplementation on blood biochemical parameters in sheep. The experiment was conducted at the FZEA/USP. Total 40 weaned sheep were divided in a completely randomized design with 10 treatments: 1) basal diet; 2) basal diet plus Mo; 3) basal diet + inorganic Cu + inorganic S; 4) basal diet + inorganic Cu + organic S; 5) basal diet + organic Cu + inorganic S; 6) basal diet + organic Cu + organic S; 7) diet plus Mo + inorganic Cu + inorganic S; 8) diet plus Mo + inorganic Cu + organic S; 9) diet plus Mo + organic Cu + inorganic S; 10) diet plus organic Cu + organic S. According each treatment was added 10mg kg-1 MS inorganic or organic Cu or 10mg kg-1 MS Mo or 0.2% S inorganic or organic. The animals were fed twice a day, a total of 3% of body weight. The experiment lasted 84 days, blood was sampled every 28 days to study glucose, urea, albumin, cholesterol and triglycerides. The parameters were analyzed with factorial structure 2 x 2 x 2 (with and without Mo, organic and inorganic Cu and organic and inorganic S) and a basal diet and a basal diet plus Mo, with 5% significance. Serum levels of glucose, urea, albumin and cholesterol showed no significant difference between treatment and time (p> 0.05). It observed means of 64.8mg dl- 1; 30.0mg dl -1; 2.78mg L -1; 72.2mg dl- 1 respectively for glucose, urea, albumin and cholesterol. Serum levels of triglycerides in the treatments (28.8; 34.8; 30.8; 36.9; 34.3; 27.0; 31.6; 32.1; 34.6; 31.1mg dl- 1) were influenced by interaction Cu x S, triglycerides decreased...

Enhancing Bio-Available Phosphorous in Soil Through Sulfur Oxidation by Thiobacilli.

Aims: The objectives were to evaluate the phosphate solubilization efficiency of different Thiobacilli strains and to find out the best combination of sulfur and Thiobacilli for enhancing bio-available P in soil. Study Design: An experimental study. Place and Duration of Study: Microbiology and Soil Fertility Labs, Department of Soil Science and Soil and Water Conservation, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan and Microbiology and Soil Chemistry Labs, Auriga Research Center, Lahore, Pakistan, between May 2011 and November 2012. Methodology: Fifty Thiobacilli strains were isolated from ten different ecologies. Then an incubation study of soil was performed wherein the most efficient four Thiobacilli strains were inoculated in combination with three different levels of elemental sulfur to determine pH, water soluble sulfur, sequential P fractions and bio-available phosphorous contents in the incubated soil. Results: All the four Thiobacillus strains (IW16, SW2, IW1 and IW14) dropped pH of the incubated soil along with three doses of S° (50, 75 and 100 kg ha-1). However, Thiobacillus strains IW16 and SW2 reduced soil pH quite sharply from 7.90 to 7.12 (net reduction of 0.78 points) and 7.28 (net reduction of 0.62 points) respectively where inoculated with S° @ 100 kg ha-1. The best P solubilizer was Thiobacillus strain IW16 and the best dose of S° was @ 100 kg ha-1 and their combination enhanced maximum quantity of P (22.26 mg kg-1) in the soil by solubilizing already present insoluble calcium bounded P fractions like octacalcium phosphate (Ca8-P) and apatite (Ca10-P). Conclusion: The present study suggests the use of Thiobacilli along with elemental sulfur for the dissolution and enhancement of bio-available P in alkaline and calcareous soils.

Modeling the kinetics of pyrite ash biodesulfurization by Saccharomyces cerevisiae and Acetobacter aceti in liquid state bioreactors

Background: Modeling the kinetics of the biodesulphurization bioprocess for the refining of pyrite ash by Saccharomyces cerevisiae and Acetobacter aceti have been studied in batch-type liquid- state bioreactors. Results: The biodesulphurization experiments were performed at varying temperatures of 25ºC, 30ºC and 35ºC for eight weeks. Glucose, acetic acid and ethyl alcohol were used in the incubation media as substrates and acid sources. pH and oxidation reduction potential (ORP) observations have been determined weekly and the dissolved sulphur was measured at the end of the eight weeks trials. An equation calculating pH was derived from the iron oxidation reaction containing the ferric to ferrous iron [Fe+3/Fe+2] ratio as a variable. The Michaelis-Menten predictive specific growth rates (qFe+2), which were estimated from pH and ORP observations, were compared by plotting [qFe+²]pH vs. [qFe+2]mV. The highest ratio of dissolved sulphur over total sulphur (Sd/St) was found to be 0.5 in the biodesulphurization processes. Conclusions: The model provides predictions of ferric to ferrous iron rates and specific growth rates [qFe+²]pH vs. [qFe+2]mV and can be used for the determination of oxidized and reduced ions. The ratios of dissolved sulphur to total sulphur (Sd/St) have shown some promising results for S. cerevisiae to be used as a biodesulphurization and refining microorganism for pyrite ash and the other sulphide minerals.

Impact of refined petroleum spills on water quality, macro-invertebrate and microbial communities of a tropical aquatic environment.

Water quality characteristics, benthic macro-invertebrates and microbial communities of three first order streams in South West Nigeria were investigated to assess the effects of refined petroleum five months after spillage. All physical and chemical conditions except temperature and pH were significantly different (P<0.01) at the upstream control stations and impacted stations reflecting the perturbational stress. The benthic macro-invertebrate fauna were dominated by arthropods, but the faunal spectrum was dissimilar at all the stations studied. Sampling stations at the epicentre of the spill showed considerable reduction in faunal compositions and relative abundance. Generally, the microbial density and diversity were highest in both soil and water samples from impacted sites than in control sites. There was a significantly higher proportion (P < 0.05) of hydrocarbon utilizers in soil than in water samples in all stations except in samples from stations (P<0.05).

General metabolism of the dimorphic and pathogenic fungus Paracoccidioides brasiliensis

Annotation of the transcriptome of the dimorphic fungus Paracoccidioides brasiliensis has set the grounds for a global understanding of its metabolism in both mycelium and yeast forms. This fungus is able to use the main carbohydrate sources, including starch, and it can store reduced carbons in the form of glycogen and trehalose; these provide energy reserves that are relevant for metabolic adaptation, protection against stress and infectivity mechanisms. The glyoxylate cycle, which is also involved in pathogenicity, is present in this fungus. Classical pathways of lipid biosynthesis and degradation, including those of ketone body and sterol production, are well represented in the database of P. brasiliensis. It is able to synthesize de novo all nucleotides and amino acids, with the sole exception of asparagine, which was confirmed by the fungus growth in minimal medium. Sulfur metabolism, as well as the accessory synthetic pathways of vitamins and co-factors, are likely to exist in this fungus.

Some morphologic and physiologic responses of some plants to hydrogen sulphide

Experiments were conducted with rocket, radish and tomato seedlings to determine the effect of H[2]S at different concentrations on the growth and some biochemical changes. H[2]S induced significant inhibitory effect on radical and plumule elongation, photosynthetic activities, accumulation of soluble sugars and biosynthesis of polysaccharides. The degrees of inhibition was a function of both concentration and duration of treatment