Influence of furfural on the physiology of Acinetobacter baylyi ADP1.

Pretreatment of lignocellulosic biomass produces growth inhibitory substances such as furfural which is toxic to microorganisms. Acinetobacter baylyi ADP1 cannot use furfural as a carbon source, instead it biotransforms this compound into difurfuryl ether using the reduced nicotinamide adenine dinucleotide (NADH)-dependent dehydrogenases AreB and FrmA during aerobic acetate catabolism. However, NADH consumption for furfural biotransformation compromises aerobic growth of A.

Transport-controlled growth decoupling for self-induced protein expression with a glycerol-repressible genetic circuit.

Decoupling cell formation from recombinant protein synthesis is a potent strategy to intensify bioprocesses. Escherichia coli strains with mutations in the glucose uptake components lack catabolite repression, display low growth rate, no overflow metabolism, and high recombinant protein yields. Fast growth rates were promoted by the simultaneous consumption of glucose and glycerol, and this was followed by a phase of slow growth, when only glucose remained in the medium.

Increasing the Pentose Phosphate Pathway Flux to Improve Plasmid DNA Production in Engineered .

The demand of plasmid DNA (pDNA) as a key element for gene therapy products, as well as mRNA and DNA vaccines, is increasing together with the need for more efficient production processes. An engineered strain lacking the phosphotransferase system and the pyruvate kinase A gene has been shown to produce more pDNA than its parental strain. With the aim of improving pDNA production in the engineered strain, several strategies to increase the flux to the pentose phosphate pathway (PPP) were evaluated.

Recombinant protein expression in proteome-reduced cells under aerobic and oxygen-limited regimes.

Industrial cultures are hindered by the physiological complexity of the host and the limited mass transfer capacity of conventional bioreactors. In this study, a minimal cell approach was combined with genetic devices to overcome such issues. A flavin mononucleotide-based fluorescent protein (FbFP) was expressed in a proteome-reduced Escherichia coli (PR).

Design of microaerobically inducible miniR1 plasmids.

Plasmid DNA manufacture is an essential step to produce gene therapy agents and next-generation vaccines. However, little attention has been paid toward developing alternative replicons that can be coupled with large-scale production conditions. Our results demonstrate that the miniR1 replicon can be efficiently induced by oxygen limitation when a copy of the regulatory protein RepA under control of a microaerobic promoter is used.

Glucose transport engineering allows mimicking fed-batch performance in batch mode and selection of superior producer strains.

Background: Fed-batch mode is the standard culture technology for industrial bioprocesses. Nevertheless, most of the early-stage cell and process development is carried out in batch cultures, which can bias the initial selection of expression systems. Cell engineering can provide an alternative to fed-batch cultures for high-throughput screening and host selection.

Assessing growth, frost tolerance, and acclimation of pine seedlings with contrasted dormancy strategies as influenced by organic nitrogen supply.

Freezing stress is a critical environmental factor affecting survival, distribution, and evolution of plants. Although there is evidence that nitrogen (N) affects frost tolerance of juvenile conifers, the magnitude and direction of such effect can diverge among species. The influence of the N source on frost tolerance has been barely studied.

Furfural biotransformation in Acinetobacter baylyi ADP1 and Acinetobacter schindleri ACE.

Objectives: To determine furfural biotransformation capabilities of Acinetobacter baylyi ADP1 and Acinetobacter schindleri ACE.

Results: Acinetobacter baylyi ADP1 and A. schindleri ACE could not use furfural as sole carbon source but when acetate was used as substrate, ADP1 and ACE biotransformed 1 g furfural/l in 5 and 9 h, respectively.

Plasmid DNA Production in Proteome-Reduced .

The design of optimal cell factories requires engineering resource allocation for maximizing product synthesis. A recently developed method to maximize the saving in cell resources released 0.5% of the proteome of by deleting only three transcription factors.

Physiological Response of W3110 and BL21 to the Aerobic Expression of .

The aerobic growth and metabolic performance of strains BL21 and W3110 were studied when the hemoglobin (VHb) was constitutively expressed in the chromosome. When VHb was expressed, acetate production decreased in both strains and was nearly eliminated in BL21. Transcriptional levels of the glyoxylate shunt genes decreased in both strains when VHb was expressed.

Drought tolerance and acclimation in Pinus ponderosa seedlings: the influence of nitrogen form.

Drought is a limiting factor to forest regeneration and restoration, which is likely to increase in intensity and duration under future climates. Nitrogen (N) nutrition is related to drought-resistance mechanisms in trees. However, the influence of chemical N form (inorganic and organic N) on physiological traits related to drought resistance has been sparsely studied in conifer seedlings.

Correction to: Gel electrophoresis of human sperm: a simple method for evaluating sperm protein quality.

[This corrects the article DOI: 10.1186/s12610-018-0076-0.].

Physiological and transcriptional comparison of acetate catabolism between Acinetobacter schindleri ACE and Escherichia coli JM101.

Acinetobacter bacteria preferentially use gluconeogenic substrates instead of hexoses or pentoses. Accordingly, Acinetobacter schindleri ACE reaches a high growth rate on acetate but is unable to grow on glucose, xylose or arabinose. In this work, we compared the physiology of A.

Design of a microaerobically inducible replicon for high-yield plasmid DNA production.

A pUC-derived replicon inducible by oxygen limitation was designed and tested in fed-batch cultures of Escherichia coli. It included the addition of a second inducible copy of rnaII, the positive replication control element. The rnaII gene was expressed from P and cloned into pUC18 to test the hypothesis that the ratio of the positive control molecule RNAII to the negative control element, RNAI, was the determinant of plasmid copy number per chromosome (PCN).

Acinetobacter baylyi ADP1 growth performance and lipid accumulation on different carbon sources.

Acinetobacter baylyi ADP1 is a microorganism with the potential to produce storage lipids. Here, a systematic study was carried out to evaluate growth performance and accumulation of wax esters and triacylglycerols using glycerol, xylose, glucose, acetate, ethanol, and pyruvate as carbon sources. High specific growth rates (μ) were found in gluconeogenic carbon sources (ethanol, acetate, and pyruvate: 0.

Engineering E. coli for improved microaerobic pDNA production.

Escherichia coli strains W3110 and BL21 were engineered for the production of plasmid DNA (pDNA) under aerobic and transitions to microaerobic conditions. The gene coding for recombinase A (recA) was deleted in both strains. In addition, the Vitreoscilla hemoglobin (VHb) gene (vgb) was chromosomally inserted and constitutively expressed in each E.

Design of a synthetic miniR1 plasmid and its production by engineered Escherichia coli.

A synthetic plasmid consisting of the minimal elements for replication control of the R1 replicon and kanamycin resistance marker, which was named pminiR1, was developed. pminiR1 production was tested at 30 °C under aerobic and microaerobic conditions in Escherichia coli W3110 recA (W1). The plasmid DNA yields from biomass (Y) were only 0.

In States That Border Mexico, Cesarean Rates Were Highest For Hispanic Women Living In Border Counties In 2015.

Hispanic women living on the US-Mexico border have had higher cesarean delivery rates than other Hispanic women in the US. Using birth certificate and other data, we compared cesarean rates among Hispanic women living in US border counties with rates among other Hispanic women in border states during 2015. Using linear regression, we also determined which medical, hospital, and sociodemographic characteristics accounted for intercounty variations in rates.

Trends in Hispanic and non-Hispanic white cesarean delivery rates on the US-Mexico border, 2000-2015.

Background: Cesarean delivery occurs in one in three US births and poses risks for mothers and infants. Hispanic cesarean rates were higher than non-Hispanic white rates in the US in 2016. In 2009, cesarean rates among Hispanics on the US-Mexico border exceeded rates among US Hispanics.

A-kinase anchor protein 4 precursor (pro-AKAP4) in human spermatozoa.

Background: A-kinase anchor protein 4 (AKAP4) and its precursor pro-AKAP4 are two major proteins in spermatozoa of rodents and mammals. Although researchers have characterized the AKAP4 expression in various species, the protein's expression in humans has not been described in detail.

Objectives: The objective of this study was to characterize human pro-AKAP4 more precisely (notably the definition of its localization and expression levels in human spermatozoa and testes).

Prospective assessment of follicular growth and the oocyte cohort after ovarian stimulation for fertility preservation in 90 cancer patients versus 180 matched controls.

A lower number of metaphase II oocytes eligible for vitrification after controlled stimulation in cancer patients has recently been reported, suggesting that cancer may impair the dynamics and quality of follicular growth. In this prospective, non-interventional study, the pattern of follicular growth and oocyte cohort after ovarian stimulation in cancer patients was analysed. Ninety cancer patients, recruited before starting chemotherapy, were compared with 180 time- and age-matched healthy controls undergoing intracytoplasmic sperm injection.

Evaluation of microbial globin promoters for oxygen-limited processes using .

Oxygen-responsive promoters can be useful for synthetic biology applications, however, information on their characteristics is still limited. Here, we characterized a group of heterologous microaerobic globin promoters in . Globin promoters from , , , , and were used to express the FMN-binding fluorescent protein (FbFP), which is a non-oxygen dependent marker.