Glutamine synthetase and protease enzyme activities and growth response of ruminal bacterium Prevotella ruminicola strain B14 to nitrogen source and concentration

Abstract

The objective of this study was to determine the effects of varying nitrogen sources and concentrations upon glutamine synthetase and protease activities in Prevotella ruminicola strain B₁4. Based on growth response it appears that ammonium chloride or pepticase limited P. ruminicola becomes nitrogen limited when nitrogen concentration is at 0.5 mM. However, when casein was provided as the sole source of nitrogen P. ruminicola becomes nitrogen limited at 2.5 mM. Glutamine synthetase activity was measured from mid‐log phase cells grown in either nitrogen‐limited or non‐limited conditions. No activity was detectable in the non‐limited treatments. However, in the N‐ limited treatments, pepticase had the highest activity (20.76 units), followed by ammonium chloride (18.72 units) and casein (14.42 units). Protease activity assays indicated that nitrogen‐limited cultures had higher proteolytic activity than non‐limited cultures. Moreover, these activities appeared to follow the same response pattern as the previously observed glutamine synthetase activities. The results of this study indicate that P. ruminicola strain B, 4 protease activity may be influenced by nitrogen concentration such that activity increases when nitrogen availability decreases.     

谷氨酰胺对鲑鱼胚胎(CHSE)细胞生长和代谢的影响

鉴于鱼类和哺乳动物的谷氨酰胺代谢途径有所不同，为了优化CHSE细胞的培养基和大量培养的过程控制。考察了谷氨酰胺对CHSE细胞生长和代谢的影响．在CHSE细胞批培养中，无谷氨酰胺的M199培养基可以支持CHSE细胞的生长，当初始谷氨酰胺浓度c=0．54mmolL^-1时，细胞可达到最高密度nmax／10^5mL^-1=16．19；而更高浓度的谷氨酰胺(c：1．76-5．62mmolL^-1)对细胞生长有抑制作用．在c=0～5．62mmolL^-1初始谷氨酰胺浓度的批培养中，葡萄糖的利用和乳酸的生成基本一致，随谷氨酰胺浓度的升高，Qglc和Qlac增加，Yn/glc降低．另外，随着初始谷氨酰胺浓度的增加(c=0～3．33mmolL^-1)，谷氨酰胺的消耗增加，Yn/gln和Yammo／gln分别下降58％和50％；当初始谷氨酰胺浓度继续增加时，谷氨酰胺的消耗、Yn/gla和Yammo/gln基本不变．此外，当初始谷氨酰胺浓度为0时，丙氨酸成为消耗性的氨基酸；Yala/gln先随初始谷氨酰胺浓度的增加(c=0．54～1．76mmolL^-1)而增加，而后又随初始谷氨酰胺浓度的增加(c=1．76～3．33mmolL^-1)而降低．最后维持不变．图4表3参16     

Variation in trace element mobility and nitrogen metabolism of Verbascum olympicum Boiss. under copper stress

The aim of this study was to evaluate the effects of copper (Cu) stress on accumulation and transport of trace elements, nitrogen assimilation, and growth parameters of Verbascum olympicum. Eight-week-old seedlings were grown in Hoagland's solution and exposed to 0, 50, 250, or 500?µM CuSO₄ for seven days in laboratory conditions. Bioaccumulation of trace elements (boron, bismuth, cobalt, Cu, iron, lithium, manganese, molybdenum, nickel, lead, zinc) in the roots and leaves was determined by inductively coupled plasma–mass spectrometry after one, three, and seven days. Chlorophyll content, nitrate reductase, and glutamine synthetase activities, soluble protein content, and biomass were determined. Copper accumulated in the roots and leaves (up to 19609.8 and 256.2?mg?kg^?1 dry weight, respectively). Other trace elements accumulated to higher levels in the roots of Cu-treated plants compared with those of control plants. High Cu concentrations decreased nitrogen-assimilatory enzyme activities. Compared with control plants, those treated with high Cu concentrations showed lower chlorophyll contents, total protein contents, and biomass. Nitrogen assimilation and growth parameters of V. olympicum were negatively affected by Cu treatment but mineral nutrition was not severely disrupted. The results support the suitability of V. olympicum as a candidate for phytoremediation of Cu-contaminated soils.     

Enhanced biohydrogen generation from organic wastewater containing NH 4 + by phototrophic bacteria Rhodobacter sphaeroides AR-3

NH ₄ ⁺ is typically an inhibitor to hydrogen production from organic wastewater by photo-bacteria. In this experiment, biohydrogen generation with wild-type anoxygenic phototrophic bacterium Rhodobacter sphaeroideswas found to be sensitive to NH ₄ ⁺ due to the significant inhibition of NH ₄ ⁺ to its nitrogenase. In order to avoid the inhibition of NH ₄ ⁺ to biohydrogen generation by R. sphaeroides, a glutamine auxotrophic mutant R. sphaeroides AR-3 was obtained by mutagenizing with ethyl methane sulfonate. The AR-3 mutant could generate biohydrogen efficiently in the hydrogen production medium with a higher NH ₄ ⁺ concentration, because the inhibition of NH ₄ ⁺ to nitrogenase of AR-3 was released. Under suitable conditions, AR-3 effectively produced biohydrogen from tofu wastewater, which normally contains 50–60 mg/L NH ₄ ⁺ , with an average generation rate of 14.2 mL/L·h. This generation rate was increased by more than 100% compared with that from wild-type R. sphaeroides.     

Effects of alcohol–kola nut interactions on brain glucose oxidase,glutamine synthase,and adenylate deaminase activities in Wistar rats

The effects of alcohol–kola nut interactions on activites of whole brain glucose oxidase, glutamine synthetase, and adenylate deaminase were examined in Wistar rats. Thirty Wistar albino rats were divided into six groups. Control group (1) received a placebo (4 mL of distilled water). Groups 2–6 were treated for a 21-day period with either 10% (v/v) alcohol, kola nut, caffeine, alcohol + kola nut, or alcohol + caffeine at 50 mg per kg in a total volume of 4 mL vehicle via gastric intubation, respectively. One day after the final treatment, the whole brain was harvested and processed to examine for several biochemical parameters, including activities of glucose oxidase, adenylate deaminase, and glutamine synthetase. Results showed that alcohol–kola nut interaction decreased the activities of glucose oxidase, adenylate deaminase, and glutamine synthetase in whole brain, suggesting inhibition of glutamine synthesis, decreased energy (ATP) production, ionic transport and decreased neuronal processes. Data indicates that alcohol–kola nut or alcohol–caffeine interactions modify neuronal processes in rat brain; however, the clinical importance of this is not known.