Co-transfer of two plasmids determining bacteriocin production and sucrose utilization in Streptococcus faecium

Abstract Streptococcus faecium strain 25 produced a bacteriocin (enterococcin Sf25), metabolized sucrose and contained three plasmids of 2.4, 4.7 and 13.0 MDa. Plasmids Sfp2.4 and Sfp4.7 were cotransferred in a filter mating procedure to sucrose negative and bacteriocin negative S. faecium strain M16. Strain M16 harboured a nonselftransferable plasmid Sfp 19.1 MDa, which was responsible for erythromycin resistance. Transcipient cells of S. faecium M16 contained the 19.1-MDa plasmid and plasmids Sfp2.4 and Sfp4.7, produced the enterococcin Sf25 and gained the ability to degrade sucrose.     

Stimulation of glucose utilization and lactate production in cultured human fibroblasts by thyroid hormone

Human dermal fibroblasts were obtained by harvesting outgrowing cells from the dermal tissue explants and cultured in Dulbecco's modified Eagle medium containing 10% fetal calf serum. After the cells reached confluency, culture was continued in the medium containing calf serum which was deprived of thyroid hormone by the treatment with activated charcoal. These fibroblasts were responsive to exogeneously added thyroid hormone (triiodothyronine) at physiological concentrations, resulting in enhanced utilization of glucose and production of lactate. This stimulation by thyroid hormone was dependent upon the length of exposure to the hormone and its concentration.The hormone did not show any effects on cellular DNA and protein content. The experimental system described above seems to be easy to reconstitute and should be useful for the elucidation of the mechanism of thyroid hormone action.     

Food utilization by insects: Interpretation of observed differences between dry weight and energy efficiencies

Over 80% of the values of approximate digestibility (AD), efficiency of conversion of assimilated food to biomass (ECD) and efficiency of conversion of ingested food (ECI) calculated using energy terms are greater than the corresponding dry weight (DW) values, based on data for over 65 species (38 studies; number of comparative values: AD=139, ECD=128 and ECI=169). Largest positive differences (energy > DW values) are 30 (AD, ECD) and 24 (ECI) percentage points and largest negative differences (energy < DW values) are 9 (AD), 11 (ECD) and 8 (ECI) percentage points. These differences generally are least for ECI (71% of the differences fall between 0 and +5 percentage points), and AD (68%), followed by ECD (only 47% fall between 0 and +5), and they may vary with temperature, food and other factors. The differences tend to increase (esp. for ECD and ECI) when comparing later with earlier instars. Energy > DW efficiency values are commonly expected for AD because of the generally greater energy content of food than feces, and for ECD and ECI because of the generally greater energy content of insect biomass than ingested and assimilated food. Deviations from predicted differences in surveyed literature data are discussed in terms of possible methodological sources of error.

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Résumé Plus de 80% des valeurs de la digestibilité approchée (AD), de l'efficacité de la conversion de la nourriture assimilée en biomasse (ECD) et de l'efficacité de la conversion de l'aliment ingéré (ECI), calculées en termes énergétiques, et obtenus à partir de données sur 65 espèces, sont supérieures aux valeurs des poids secs correspondants (DW): 38 études; valeurs comparatives: AD=139, ECD=128, ECI=169. Les plus importantes différences positive (énergie>valuers DW) sont de 30 (AD, ECD) et de 24 (ECI) centièmes (les différences négatives les plus fortes = 9 (AD), 11 (ECD) et 8 (ECI); ces différences sont moindres pour ECI (71% des différences tombent à 0 et +5 centièmes), et AD (68%), suivi de ECD (seulement 47% tombent entre 0 et +5). Ces différences peuvent varier avec la température, l'alimentation et d'autres facteurs; les différences tendent à croître (particulièrement pour ECD et ECI) quant on les compare plus tard avec des stades plus précoces. Energie > aux valeurs d'efficacité DW sont généralement attendues pour AD par suite du contenu énergétique supérieur de l'aliment à celui des excréments, et pour ECD et ECI par suite du contenu énergétique généralement plus élevé pour la biomasse de l'insecte que pour l'aliment ingéré et assimilé. Les écarts par rapport aux différences prédites dans les données de la littérature examinée sont analysées en considérant les sources possibles d'erreurs méthodologiques.

     

Degradation of Machine Oil by Nocardioform Bacteria

Gas liquid chromatography (GLC) was used for the first time to screen for machine oil–degrading microorganisms. Oil degradation was evaluated from the microorganism respiratory activity during the utilization of oil as the sole carbon and energy source. The results are consistent with those obtained by the conventional weighing method. Substrate specificity of the active strains with respect to different machine oils was studied. Bacterial communities exhibited the highest activity, whereas a Rhodococcus erythropolisstrain was the most active among pure cultures. Various stages of bacterial interaction with oil drops were followed by means of fluorescent microscopy.     

Künstliche Luzerneheubereitungsverfahren in Ungarn

Abstract

An experiment was performed with Rumen Simulation Technique (Rusitec) in which the fermentation of mixed ration of hay (12.8 g/d) and wheat bran (3.2 g/d) was compared with the fermentation of the same diet supplemented with 2.5 and 10 mg monensin. The duration of the experiment was 12 days. During the first six days the fermentation conditions in Rusitec were stabilised. The end products of fermentation and indices of protein synthesis were determined. The energy efficiency of volatile fatty acids (VFA), proportion of fermented hexose energy on VFA energy and on energy of bacterial matter were increased in the presence of monensin. The proportion of fermented hexose energy in methane energy was decreased. Utilization of glucose and production of adenosintriphosphate were not affected. The balance of metabolic H₂ was reduced and this was manifested in decreasing production, utilization and recovery of metabolic H₂. Microbial efficiency expressed per mol ATP (g/mol) was increased by monensin from 7.8 to 12.6. The indices of protein synthesis — protein conversion ratio, degradability of protein and microbial protein synthesis efficiency were increased and non protein utilization ratio decreased in the presence of monensin.

Es wurde ein Versuch nach der Pansensimulationsmethode durchge-führt, in dem die Fermentierung einer gemischten Ration aus Heu (12,8 g/d) und Weizenkleie (3,2 g/d) mit der Fermentierung der gleichen Ration unter Zusatz von 2, 5 und 10 mg Monensin/d verglichen wurde. Die Dauer des Versuchs betrug 12 Tage, wovon die ersten sechs Tage der Stabilisierung der Fermentierungsbedingungen im Rusitec-System dienten.

Es wurden die Endprodukte der Fermentierung und die Kennziffern der Proteinsynthese bestimmt. Die Energieausbeute an Flüchtigen Fettsäuren (FFS), der Energieanteil fermentierter Hexose an der FFS-Energie und der Energie an Bakterienmasse erhöhten sich bei Zusatz von Monensin. Der Energieanteil fermentierter Hexose an der Methanenergie verringerte sich. Die Ausnutzung von Glukose und die Produktion von Adenosintriphosphat waren unbeeinflußt. Die Bilanz von metabolischem H₂ war verringert, was sich im Absinken der Production, der Nutzung und der Wiederauffindung von metabolischem H₂ zeigte. Die mikrobielle Wirksamkeit pro mol ATP (g/mol) erhöhte sich durch Monensin von 7,8 auf 12,6. Die Kennziffern der Proteinsynthese, die Proteinkonversionsrate, die Abbaubarkeit von Protein und die mikrobielle Proteinsynthesewirksamkeit erhöhten sich und die NPN-Nutzungsrate verringerte sich in Gegenwart von Monensin.     

Synergetic utilization of glucose and glycerol for efficient myo-inositol biosynthesis

myo-Inositol (MI) as a dietary supplement can provide various health benefits. One major challenge to its efficient biosynthesis is to achieve proper distribution of carbon flux between growth and production. Herein, this challenge was overcome by synergetic utilization of glucose and glycerol. Specifically, glycerol was catabolized to support cell growth while glucose was conserved as the building block for MI production. Growth and production were coupled via the phosphotransferase system, and both modules were optimized to achieve efficient production. First, the optimal enzyme combination was established for the production module. It was observed that enhancing the production module resulted in both increased MI production and better cell growth. In addition, glucose was shown to inhibit glycerol utilization via carbon catabolite repression and the inhibition was released by over-expressing glycerol kinase. Furthermore, the inducible promoter was replaced by strong constitutive promoters to avoid inducer use. With these efforts, the final strain produced MI with both high titer and yield. In fed-batch cultivation, 76 g/L of MI was produced, showing scale-up potential. This study provides a promising strategy to achieve rational distribution of carbon flux.     

Self-assembling Shell Proteins PduA and PduJ have Essential and Redundant Roles in Bacterial Microcompartment Assembly

Protein self-assembly is a common and essential biological phenomenon, and bacterial microcompartments present a promising model system to study this process. Bacterial microcompartments are large, protein-based organelles which natively carry out processes important for carbon fixation in cyanobacteria and the survival of enteric bacteria. These structures are increasingly popular with biological engineers due to their potential utility as nanobioreactors or drug delivery vehicles. However, the limited understanding of the assembly mechanism of these bacterial microcompartments hinders efforts to repurpose them for non-native functions. Here, we comprehensively investigate proteins involved in the assembly of the 1,2-propanediol utilization bacterial microcompartment from Salmonella enterica serovar Typhimurium LT2, one of the most widely studied microcompartment systems. We first demonstrate that two shell proteins, PduA and PduJ, have a high propensity for self-assembly upon overexpression, and we provide a novel method for self-assembly quantification. Using genomic knock-outs and knock-ins, we systematically show that these two proteins play an essential and redundant role in bacterial microcompartment assembly that cannot be compensated by other shell proteins. At least one of the two proteins PduA and PduJ must be present for the bacterial microcompartment shell to assemble. We also demonstrate that assembly-deficient variants of these proteins are unable to rescue microcompartment formation, highlighting the importance of this assembly property. Our work provides insight into the assembly mechanism of these bacterial organelles and will aid downstream engineering efforts.