Influence of impeller speed on citric acid production and selected enzyme activities of the TCA cycle

Summary The effect of impeller speed on citric acid production and selected enzyme activities of the TCA cycle was studied. The highest yield of citric acid (28 g/l) was obtained in culture agitated at lower speed (300 rpm). The activity of citrate synthase decreased with the increase of speed of agitation, while the activity of aconitase and isocitrate dehydrogenase increased with the increase in agitation speed.     

Kinetics of lactate fermentation and citrate bioconversion by Lactococcus Iactis ssp. Iactis in batch culture

The growth kinetics of Lactococcus lactis ssp. lactis were studied in batch culture in conditions of non-limiting lactose and the presence of citric acid. The control of pH modified growth and citrate metabolism but did not change the yield of acid formation. At controlled pH the growth rate was unaffected by citrate metabolism. Lactose was transformed to L-lactate and assay of the metabolic by-products showed some heterofermentation at the end of the growth of cultures with low growth rates. This heterofermentation was interpreted as a slowing down of glycolysis with activation of both the pyruvate formate lyase (PFL) and the pyruvate dehydrogenase complex (PDHC). Under these conditions the presence of citric acid affected the activity of both the PDHC and the alcohol dehydrogenase (ADH). L-Lactate remains the major fermentation end-product and the sole inhibitor of fermentation, this inhibition was greater on growth than on lactic acid production.     

Effects of extremely low-frequency magnetic field on growth and differentiation of human mesenchymal stem cells

Human Mesenchymal Stem Cells (hMSCs) were exposed to a developed extremely low-frequency (ELF) magnetic fields (50?Hz ,20?mT ELF) system to evaluate whether exposure to (ELF) magnetic fields affects growth, metabolism, and differentiation of hMSCs. MTT method was used to determine the growth and metabolism of hMSCs following exposure to ELF magnetic fields. Na(+)/K(+) concentration and osmolality of extracellular were measured after exposured culture. Alkaline phosphatase (ALP) assay and Calcium assay, ALP staining, and Alizarin red staining were performed to evaluate the osteogenic differentiation of hMSCs under the ELF magnetic field exposure. In these experiments, the cells were exposed to ELF for up to 23 days. The results showed that exposure to ELF magnetic field could inhibit the growth and metabolism of hMSC, but have no significant effect on differentiation of hMSCs. These results suggested that ELF magnetic field may influence the early development of hMSCs related adult cells.     

草酸青霉纤维素酶基因和木聚糖酶基因的调控基因POX05145的鉴定

草酸青霉能产生完整的纤维素酶和木聚糖酶酶系,其纤维素酶基因的表达主要受转录因子的调控。前期工作中,通过对草酸青霉菌株HP7-1在不同碳源培养基培养条件下转录组的比较分析,获得了调控纤维素酶和木聚糖酶产量的候选调控基因集。本研究以草酸青霉ΔPoxKu70为出发菌株,通过同源重组法,构建并获得了其中一个候选调控基因POX05145的缺失突变株ΔPOX05145。在微结晶纤维素Avicel诱导培养条件下,与出发菌株ΔPoxKu70相比,ΔPOX05145的纤维素酶产量和木聚糖酶产量发生了显著改变。其中,在诱导第2天时,ΔPOX05145对硝基苯-β-D-纤维二糖苷酶产量和木聚糖酶产量分别上升43.4%和164.7%,对硝基苯-β-D-半乳糖吡喃葡萄糖苷酶产量下降92.8%,但是,滤纸酶产量和羧甲基纤维素酶产量没有显著变化。然而,在诱导第4天时,所有纤维素酶产量和木聚糖酶产量上升100.4%~294.0%。实时荧光定量PCR检测表明POX05145在不同的时间不同程度的调控主要的纤维素酶基因和木聚糖酶基因的表达。序列分析表明POX05145含有一个GAL4类锌指结构的DNA结合功能域和一个保守的真菌特有的转录因子结构域(Fungal_TF_MHR)。     

Effects of Extremely Low-Frequency Magnetic Field on Growth and Differentiation of Human Mesenchymal Stem Cells

Human Mesenchymal Stem Cells (hMSCs) were exposed to a developed extremely low-frequency (ELF) magnetic fields (50?Hz ,20?mT ELF) system to evaluate whether exposure to (ELF) magnetic fields affects growth, metabolism, and differentiation of hMSCs. MTT method was used to determine the growth and metabolism of hMSCs following exposure to ELF magnetic fields. Na⁺/K⁺ concentration and osmolality of extracelluar were measured after exposured culture. Alkaline phosphatase (ALP) assay and Calcium assay, ALP staining, and Alizarin red staining were performed to evaluate the osteogenic differentiation of hMSCs under the ELF magnetic field exposure. In these experiments, the cells were exposed to ELF for up to 23 days. The results showed that exposure to ELF magnetic field could inhibit the growth and metabolism of hMSC, but have no significant effect on differentiation of hMSCs. These results suggested that ELF magnetic field may influence the early development of hMSCs related adult cells.     

Oxygen requirements for growth and citric acid production of Yarrowia lipolytica

During continuous cultivation of Yarrowia lipolytica N 1, oxygen requirements for growth and citric acid synthesis were found to depend on the iron concentration in the medium. A coupled effect of oxygen and iron concentrations on the functioning of the mitochondrial electron transport chain in Y. lipolytica N 1 was established. Based on the results obtained in continuous culture, conditions for citric acid production in a batch culture of Y. lipolytica N 1 were proposed. At relatively low pO(2) value and a high iron concentration, citric acid accumulation was as high as 120 g l(-1); the specific rate of citric acid synthesis reached 120 mg citric acid (g cells h)(-1). The mass yield coefficient was 0.87 and the energy yield coefficient was 0.31.     

Influence of nitrilotriacetic acid and 8-hydroxyquinoline on the production of citric acid from molasses using Aspergillus niger

The effect of nitriloacetic acid (NTA) and 8-hydroxyquinoline on the production of citric acid by Aspergillus niger was investigated. The complexing agents showed an effect only during inoculation of the microorganism. Subsequent addition after inoculation did not produce any significant increase in citric acid yield. When 200 ppm of NTA was added during inoculation, an increase of 10.9 g·dm⁻³ citric acid over that produced by the control culture was observed. 8-Hydroxyquinoline, on the other hand, produced a higher concentration of citric acid which was 34 g·dm⁻³ over that of the control culture. The use of 8-hydroxyquinoline is therefore suggested for the production of citric acid from molasses using Aspergillus niger.     

Monitoring the inhibitive effect of the static magnetic field on the activity of lysozyme with acoustic wave impedance analysis technique

The inhibitive effect of static magnetic field on the activity of lysozyme was studied using acoustic wave impedance analysis technique. Equivalent circuit parameters of piezoelectric quartz crystal (PQC) were obtained and discussed. The results showed that the activity of lysozyme was inhibited due to the effect of static magnetic field and the inhibitive effect becomes greater with an increase in magnetization time or magnetic field intensity. According to the response characteristics of motional resistance change (deltaR1), which is related to the change in the bacterial number, a quantitative response model reflecting the activity of lysozyme was theoretically derived. By fitting deltaR1 versus time curves under a specific magnetic field intensity but different magnetic time to the model, the relationship between K1 reflecting the activity of lysozyme and magnetic time t(m) was established. Based on the relationship, a new impedance response model that indicates the inhibitive influence of the magnetization time on the activity of lysozyme was derived as follows: deltaR1 = R0((K4(exp[K0exp(-0.26t(m))]t - 1) + 1)1/2 - 1). Similarly, another response model that indicates the effect of magnetic field intensity was derived as follows: deltaR1 = R0((K4(exp(K0 exp(- 5.17B)t) - 1) + 1)1/2 - 1).     

Seed pretreatment with magnetic field alters the storage proteins and lipid profiles in harvested soybean seeds

The increase in crop productivity is an urgent need of the time to reduce scarcity of food in underdeveloped countries. Several biological, chemical and physical methods have been applied to promote crop yield. Application of magnetic field (MF) is an emerging physical method used to increase plant growth and yield. The reports on MF pretreatment-induced nutritional changes in harvested seeds are scarce. We previously identified the optimal frequency of MF to improve plant growth and yield as 1500 nT at 10.0 Hz. This study was aimed to investigate the effect of MF treatment on storage proteins and fatty acids in harvested soybean seeds. The results showed that MF triggered globulin production and suppressed prolamin production. However, lipid content in seeds increased, because MF exposure caused an elevation of several fatty acids including caprylic acid, palmitic acid, heptadecanoic acid, linoleic acid, lignoceric acid and eicosapentaenoic acid. This is the first report to reveal the seed pretreated MF on nutritional values of harvested seeds. This study suggests that MF treatment improves seed quality by regulating the metabolism of storage proteins and fatty acids.     

Activity of acid and alkali phosphatase in guinea pigs exposed to the static magnetic fields

The influence of static homogeneous magnetic field on alkali and acid phosphatase in guinea pigs (regarding the twenty four hours rhythm) was studied. The increase of acid phosphatase activity was determined by the time of exposure to magnetic field. No changes in alkali phosphatase activity were observed.     

Simultaneous saccharification and fermentation of cellulose: Effect of ethanol and cellulases on particular stages

The effects of ethanol and Trichoderma reesei cellulase on the saccharification and fermentation processes as well as the tolerance of the cellulase complex for ethanol have been investigated. The studies were conducted with respect to their usefulness in the process of simulataneous saccharification and fermentation of cellulose to ethanol. The following results were obtained. (1) Fermentative activity of Kluyveromyces fragilis yeasts was gradually depressed with increasing intial ethanol concentrations and temperature of fermentation between 35–46°C. (2) Crude cellulase preparation introduced to the culture broth to a final enzyme activity of 0.5 to 2.0 FPU/ml had not distinct effect on the biomass production, ethanol yield, and glucose uptake by yeasts in 48 h fermentation at 43°C. On the other hand, only a negligible decrease in the cellulase complex activity was observed during fermentation process. (3) Saccharification of wheat straw was inhibited by at least 1% w/v ethanol. (4) The enzymes of the cellulase system showed a high stability to exposure to ethanol for 48 h at 43°C.     

Modulation of collagen production in cultured fibroblasts by a low-frequency, pulsed magnetic field

Primary cultures of chicken tendon fibroblasts have been exposed for various periods to a low-frequency, pulsed magnetic field, and the effects on protein and collagen synthesis have been examined by radioisotopic incorporation. Total protein synthesis was increased in confluent cells treated with a pulsed magnetic field for the last 24 h of culture as well as in cells treated for a total of 6 days. However, in 6 day-treated cultures, collagen accumulation was specifically enhanced as compared to total protein, whereas after short-term exposure, collagen production was increased only to the same extent as total protein. Levels of cyclic AMP were significantly decreased after 6-day pulsed magnetic field treatment, probably as a consequence of diminished adenylate cyclase activity. Exposure to pulsed magnetic field had no effect on cell proliferation or collagen phenotype. These results indicate that a pulsed magnetic field can specifically increase production of collagen, the major differentiated function of fibroblasts, possibly by altering cyclic-AMP metabolism.     

Metabolic effects of static magnetic fields on Streptococcus pyogenes

This study aimed to develop a simple experimental system utilising bacterial cells to investigate the dose responses resulting from exposures to static magnetic flux densities ranging from 0.05 to 0.5 T on viability, bacterial metabolism and levels of DNA damage in Streptococcus pyogenes. Exposure of S. pyogenes to a field of 0.3 T at 24 degrees C under anaerobic conditions resulted in a significant (P < 0.05) decrease in growth rate, with an increased mean generation time of 199 +/- 6 min compared to the control cells at 165 +/- 6 min (P < 0.05). Conversely, exposure to magnetic fields of 0.5 T significantly accelerated the growth rate at 24 degrees C compared to control cells, with a decreased mean generation time of 147 +/- 4 min (P < 0.05). The patterns of metabolite release from cells incubated in phosphate buffered saline (PBS) at 24 degrees C and exposed to different magnetic flux densities (0.05-0.5 T) were significantly (P < 0.05) altered, compared to non-exposed controls. Concentrations of metabolites, with the exception of aspartic acid (r = 0.44), were not linearly correlated with magnetic flux density, with all other r < 0.20. Instead, "window" effects were observed, with 0.25-0.3 T eliciting the maximal release of the majority of metabolites, suggesting that magnetic fields of these strengths had significant impacts on metabolic homeostasis in S. pyogenes. The exposure of cells to 0.3 T was also found to significantly reduce the yield of 8-hydroxyguanine in extracted DNA compared to controls, suggesting some possible anti-oxidant protection to S. pyogenes at this field strength.     

植酸钠对黑曲霉柠檬酸发酵产酸的促进效应

研究了植酸钠对黑曲霉柠檬酸发酵产酸的促进效应。在葡萄糖全合成培养基中添加１％的植酸钠,可使产酸比对照提高２．４倍;在薯粉、玉米粉等天然培养基中添加１％植酸钠,柠檬酸产量分别提高１．７倍和１．３倍。酶活性测定分析表明,植酸钠对柠檬酸代谢途径中的几种关键酶的活性有影响。     

Cellulase production in continuous culture by Trichoderma reesei on xylose-based media

Trichoderma reesei (QM 9414) produced cellulase in continuous culture, on media containing xylose (1%) supplemented with sorbose (0.3%) to induce cellulase production. Maximum cell mass of 4.54 kg/m(3) occurred at pH 4.0 and a dilution rate of 0.0391 h(-1) where residual substrate was 0.43 kg/m(3), but no cellulase was produced. Maximum cellulase production of 0.69 FPU occurred at pH 3.5 and a dilution rate of 0.0110 h(-1), where cell mass production was 2.56 kg/m(3) and residual substrate was 0.15 kg/m(3). Monod kinetic constants, corrected for endogenous metabolism, were 0.091 h(-1), 0.469 kg/m(3), 0.00923 h(-1), and 0.470 kg cells/kg xylose at pH 3.5, for the maximum specific growth rate, Michaelis-Menten coefficient, endogenous metabolism coefficient, and yield coefficient, respectively. Specific growth rate fitted a maturation time model, which predicted decreasing maturation time with increasing pH.     

Chronic exposure to a 2 mT static magnetic field affects the morphology,the metabolism and the function of in vitro cultured swine granulosa cells

In recent years, the exposure of organisms to static magnetic fields (SMFs) is continuously increasing. Thus, we investigated the effect of chronic exposure to a 2 mT SMF on in vitro cultured swine granulosa cells (GCs). In particular, the culture expansion (cell viability and doubling time), the cell phenotype (cell morphology and orientation, actin and α-tubulin cytoskeleton), the cell metabolism (intracellular Ca²⁺ concentration [Ca²⁺]_i and mitochondrial activity) and the cell function (endocrine activity) were assessed. It has been found that the exposure to the field did not affect the cell viability, but the doubling time was significantly reduced (p < 0.05) in exposed samples after 72 h of culture. At the same time, the cell length and thickness significantly changed (p < 0.05), while the cell orientation was unaffected. Evident modifications were induced on actin and α-tubulin cytoskeleton after 3 days of exposure and, simultaneously, a change in [Ca²⁺]_i and mitochondrial activity started to become evident. Finally, the SMF exposure of GCs longer than 72 h determined a significant alteration of progesterone and estrogen production (p < 0.05). In conclusion, our results demonstrate that the chronic exposure of swine GCs to a 2 mT SMF exerts a negative effect on cell proliferation, morphology, biochemistry and endocrine function in an in vitro model.     

Biosynthesis of citric acid by Yarrowia lipolytica repeat-batch culture on ethanol

After analysis of batch culture and identification of the ways for prolongation of citric acid active synthesis by yeast, repeat-batch (RB) cultivation was suggested. Yarrowia lipolytica strain RB cultivation was studied and optimal conditions for cultivation selected. It was shown that when applying RB cultivation, better results were obtained than for batch cultivation. The activity of the culture remained stable after cultivation for more than 700 h. Comparative analysis of enzyme activities confirmed the regularity of the effect described, as the activity of practically of all the enzymes participating in ethanol oxidation and citric acid biosynthesis remained stable over time during RB cultivation. Advantages of RB cultivation for the production of citric acid by yeast are discussed. Received: 1 March 1999 / Received revision: 28 June 1999 / Accepted: 5 July 1999     

Retinoic acid blockade of imidazole-induced tyrosinase expression in B16 melanoma cultures: similar effects of the active retinoid and triiodothyronine

The effect of retinoic acid on the induction of tyrosinase (EC 1:14.18.1) by imidazole was determined in cultured B16/C3 melanoma cells. Retinoic acid could block the induction of enzyme activity within 3 hours of addition to the culture medium at a physiological concentration (10nM). The blockade was similar to that of 3,3',5-L-triiodothyronine (T3) already reported. mRNA hybridizable to a tyrosinase DNA probe was induced by imidazole while retinoic acid and T3 blocked that increase. These observations suggest that retinoic acid can mimic the action of T3 in B16 melanoma cells in culture.     

Effect of magnetic field on the accumulation of polyhydroxyalkanoates (PHAs) by microorganism in activated sludge

The effect of static magnetic field on polyhydroxyalkanoates (PHAs) syntheses by activated sludge under aerobic dynamic feeding (ADF) was evaluated in sequence batch reactors (SBR), with magnetic field intensities of 42, 21, 11 and 7 millitesla (mT) exposure in the feast, feast-famine and famine periods, respectively, and one control group without magnetic field exposure. Under each level of magnetic field intensity, the effect of magnetic field exposed in the famine period to PHAs syntheses was most significant in comparison with that in the feast or feast-famine period. Maximal hydroxybutyrate (HB) and (HV) yield occurred at 21 and 11 mT, respectively, and the minimal yield occurred at 42 mT during exposure in the famine period. The maximum biodegradable rate constant of PHA was noted at 11 mT during exposure in the famine period.