水体pH胁迫对异育银鲫皮质醇激素和非特异性免疫的影响

实验比较了不同pH胁迫对异育银鲫(Carassius auratus gibelio)皮质醇激素和非特异性免疫的影响。3个实验组水质的pH分别为6.0、8.5、9.5,对照组是pH为7.4的曝气自来水。分别在实验的第0、14和30天采血,血浆用于测定皮质醇含量和溶菌酶活性;分离的白细胞,采用流式细胞术测定吞噬活性和呼吸暴发。在实验的第4天多取1次血样,用于皮质醇含量测定。在实验的第31天,测量鱼的体重,并用柱状黄杆菌(Flavobacterium columnare)浸泡攻毒,7d后统计死亡情况。结果显示:在pH 6.0的胁迫下,白细胞的吞噬活性在第14、30天显著降低,而呼吸暴发和皮质醇含量没有显著变化,溶菌酶活性和生长与对照组没有显著差异,用黄杆菌攻毒也没有出现死亡。而在pH 8.5和9.5的胁迫下,皮质醇含量在第14、30天显著升高,白细胞的吞噬活性、呼吸暴发显著降低;在pH 9.5胁迫下的溶菌酶活性显著降低,用柱状黄杆菌攻毒两组都出现死亡。实验表明pH为8.5和9.5的胁迫严重影响了异育银鲫的非特异性免疫功能,流式细胞术能较客观测定白细胞的吞噬活性和呼吸暴发,可用于评价胁迫对鱼类健康的影响。     

Phenylphosphonate transport by Helicobacter pylori

BACKGROUND: Helicobacter pylori can utilize phenylphosphonate as a sole source of phosphorus, and it is able to transport the phosphonate N-phosphonoacetyl-L-aspartate. However, H. pylori does not have any genes homologous to those of the known pathways for phosphonate degradation in bacteria, indicating that it must have novel pathways for the transport and metabolism of phosphonates. METHODS: Phenylphosphonate transport by H. pylori was studied in strains LC20, J99 and N6 by the centrifugation through oil method using [(14)C]-labeled phenylphosphonate. RESULTS: The Michaelis constants of transport K(t) and V(max) for phenylphosphonate showed similar kinetics in the three strains. The Arrhenius plot for phenylphosphonate transport rates at permeant concentrations of 50 micromol/L was linear over the temperature range 10-40 degrees C with an activation energy of 3.5 kJ/mol, and a breakpoint between 5 and 10 degrees C. Transport rates increased with monovalent cation size. The effects of various inhibitors were investigated: iodoacetamide, amiloride, valinomycin, and nigericin reduced the rate of phenylphosphonate transport; sodium azide and sodium cyanide increased the transport rate; and monensin had no effect. CONCLUSIONS: The kinetics and properties of H. pylori phenylphosphonate transport were characterized, and the data suggested a carrier-mediated transport mechanism.     

Mass balance of heavy metal uptake by encapsulated cultures ofKlebsiella aerogenes

Dialysis was employed as a method of speciating heavy metals in cultures of an extracellular polymer forming strain ofKlebsiella aerogenes. A noncapsulated strain of the same bacterium was used as a control, and a mass balance of copper, cadmium, cobalt, nickel, and manganese in batch culture at pH 4.5 and pH 6.8 and in continuous culture at pH 6.8 was constructed. Copper and cadmium were accumulated by the cell during rapid proliferation whereas all 5 metals were bound nonspecifically by extracellular polymer produced during stationary phase and at low dilution rates. The presence of extracellular polymer appeared to inhibit cellular uptake of nickel. At the lower pH, metal uptake was considerably reduced. The results are discussed in the context of metal removal in the activated sludge process of waste water treatment.     

The effects of pH and salts on nucleic acid partitioning during phenol extraction

The partitioning of nucleic acids is sensitive to pH during phenol extraction. However, the exact effects of pH on phenol extraction had not been systematically investigated, and the mechanism of which were not fully elucidated. In this paper, we showed that the partitioning of nucleic acids was determined neither solely by the pH of the aqueous buffer being used, nor by the “pH of the phenol”; the latter is a completely wrong conception. We demonstrated that a key determinant for nucleic acid partitioning during phenol extraction was the equilibrated pH of the aqueous phase, which should be defined as the pH of phenol extraction. For example, when 50?mM NaAc-HAc buffer at pH of 3.47 was mixed with an equal volume of water-saturated phenol, the equilibrated pH of aqueous phase would be raised to ～3.84. At this pH, almost all of genomic DNA partitioned into the phenol phase, and genomic DNA-free total RNA was retained in the aqueous phase. Several salts were found affecting the partitioning of nucleic acids during phenol extraction in different manners. Based on these results, a low-cost and efficient method for genomic DNA-free total RNA extraction was developed.     

Mannitol production by heterofermentative <Emphasis Type="BoldItalic">Lactobacillus reuteri</Emphasis> CRL 1101 and <Emphasis Type="BoldItalic">Lactobacillus fermentum</Emphasis> CRL 573 in free and controlled pH batch fermentations

Certain lactic acid bacteria, especially heterofermentative strains, are capable to produce mannitol under adequate culture conditions. In this study, mannitol production by Lactobacillus reuteri CRL 1101 and Lactobacillus fermentum CRL 573 in modified MRS medium containing a mixture of fructose and glucose in a 6.5:1.0 ratio was investigated during batch fermentations with free pH and constant pH 6.0 and 5.0. Mannitol production and yields were higher under constant pH conditions compared with fermentations with free pH, the increase being more pronounced in the case of the L. fermentum strain. Maximum mannitol production and yields from fructose for L. reuteri CRL 1101 (122 mM and 75.7 mol%, respectively) and L. fermentum CRL 573 (312 mM and 93.5 mol%, respectively) were found at pH 5.0. Interestingly, depending on the pH conditions, fructose was used only as an alternative external electron acceptor or as both electron acceptor and energy source in the case of the L. reuteri strain. In contrast, L. fermentum CRL 573 used fructose both as electron acceptor and carbon source simultaneously, independently of the pH value, which strongly affected mannitol production by this strain. Studies on the metabolism of these relevant mannitol-producing lactobacilli provide important knowledge to either produce mannitol to be used as food additive or to produce it in situ during fermented food production.     

Influence of Transglutaminase (TGase) Enzyme on Mechanical and Bioactive Properties of Crayfish Protein Gels

Functional and bioactive properties of crayfish meat convert their surpluses in an excellent alternative for the development of food products. Thus, protein dispersions were subjected to a thermal treatment, obtaining a protein-based gel. Rheological and antioxidant properties were studied at three different pH values (2.0, 6.5 and 8.0) when the TGase enzyme was used. The pH value exerted a strong influence on the gelation behaviour, as well as on the functional properties and the antioxidant activity of the final gels. The activity of the TGase enzyme is highly influenced by the pH of the protein dispersions. The highest antioxidant activity was obtained against ABTS and the lowest when FC reagent was used, whereas the activity against DPPH was also remarkable. TGase enzyme can be used during the thermal treatment to increase the mechanical properties, which were lost when hydrolysate systems were used.     

P NMR Observations on the Effect of the External pH on the Intracellular pH Values in Plant Cell Suspension Cultures

³¹P nuclear magnetic resonance (NMR) spectroscopy was used to monitor the response of oil palm (Elaeis guineensis) and carrot (Daucus carota) cell suspensions to changes in the external pH. An airlift system was used to oxygenate the cells during the NMR measurements and a protocol was developed to enable a constant external pH to be maintained in the suspension when required. Phosphonoacetic acid was used as an external pH marker and the intracellular pH values were measured from the chemical shifts of the cytoplasmic and vacuolar orthophosphate resonances. In contrast to earlier studies the cytoplasmic pH was independent of the external pH over the range 5.5 to 8.0 and it was only below pH 5.5 that the cytoplasmic pH varied, falling at a rate of 0.12 pH unit per external unit. Loss of pH control was observed in response to sudden increases in external pH with the response of the cells depending on the conditions imposed. A notable feature of the recovery from these treatments was the transient acidification of the cytoplasm that occurred in a fraction of the cells and overshoot phenomena of this kind provided direct evidence for the time dependence of the regulatory mechanisms.     

Bioreductive activation of mitomycin C by DT-diaphorase.

The role of DT-diaphorase (DTD, EC 1.6.99.2) in the bioreductive activation of mitomycin C was examined using purified rat hepatic DTD. The formation of adducts with reduced glutathione (GSH), binding of [3H]mitomycin C to DNA, and mitomycin C-induced DNA interstrand cross-linking were used as indicators of bioactivation. Mitomycin C was metabolized by DTD in a pH-dependent manner with increasing amounts of metabolism observed as the pH was decreased from 7.8 to 5.8. The major metabolite observed during DTD-mediated reduction of mitomycin C was 2,7-diaminomitosene. GSH adduct formation, binding of [3H]mitomycin C and mitomycin C-induced DNA interstrand cross-linking were observed during DTD-mediated metabolism. In agreement with the pH dependence of metabolism, increased bioactivation was observed at lower pH values. Temporal studies and experiments using authentic material showed that 2,7-diaminomitosene could be further metabolized by DTD resulting in the formation of mitosene adducts with GSH. DNA cross-linking during either chemical (sodium borohydride) or enzymatic (DTD) mediated reduction of mitomycin C could be observed at pH 7.4, but it increased as the pH was decreased to 5.8, showing the critical role of pH in the cross-linking process. These data provide unequivocal evidence that the obligate two-electron reductase DTD can bioactivate mitomycin C to reactive species which can form adducts with GSH and DNA and induce DNA cross-linking. The use of mitomycin C may be a viable approach to the therapy of tumors high in DTD activity, particularly when combined with strategies to lower tumor pH.     

Characteristics of film based on protein isolate from red tilapia muscle with negligible yellow discoloration

The properties of film from fish protein isolate (FPI) prepared by prior washing followed by alkaline solubilization process (ASP) from red tilapia muscle were monitored during the storage of 20 days at 50% RH and 25°C, in comparison with those of films from washed mince. Lipid, heme iron and non-heme iron contents in FPI were decreased by 98.8, 36.8 and 91.9%, respectively in comparison with those of washed mince (p<0.05). Films from FPI had higher tensile strength (TS) and elongation at break (EAB) than those from washed mince for both pH (3 and 11) used for film preparation (p<0.05). Film from FPI at pH 3 showed the highest TS, while that from washed mince at pH 11 had the lowest TS (p<0.05). Nevertheless, films from FPI had higher WVP than those from washed mince for both pH used (p<0.05). At the same pH used for film preparation (3 or 11), films from FPI showed the lower TBARS values than those from washed mince (p<0.05). Nevertheless, films from both FPI and washed mince had the higher TBARS values when pH 3 was used for film preparation, compared with pH 11 (p<0.05). Among all films, those from FPI prepared at pH 3 had the highest transparency and no yellow discoloration was observed during the storage of 20 days, in comparison with other films (p<0.05). Conversely, film from washed mince prepared at pH 3 had the higher increase in b*-value and ΔE*-value than other films. Therefore, FPI could serve as a potential material for film preparation with lower contents of lipid and prooxidants, thereby preventing the yellow discoloration of the fish myofibrillar protein-based film during extended storage.     

Online recovery of nisin during fermentation and its effect on nisin production in biofilm reactor

An online removal of nisin by silicic acid coupled with a micro-filter module was proposed as an alternative to reduce detrimental effects caused by adsorption of nisin onto producer, enzymatic degradation by protease, and product inhibition during fermentation. In this study, silicic acid was successfully used to recover nisin from the fermentation broth of Lactococcus lactis subsp. lactis NIZO 22186. The effect of pH (at 6.8 and 3.0) during adsorption process and several eluents (deionized water, 20% ethanol, 1 M NaCl, and 1 M NaCl + 20% ethanol) for desorption were evaluated in a small batch scale. Higher nisin adsorption onto silicic acid was achieved when the adsorption was carried out at pH 6.8 (67% adsorption) than at pH 3.0 (54% adsorption). The maximum recovery was achieved (47% of nisin was harvested) when the adsorption was carried out at pH 6.8 and 1 M NaCl + 20% ethanol was used as an eluent for desorption. Most importantly, nisin production was significantly enhanced (7,445 IU/ml) when compared with the batch fermentation without the online recovery (1,897 IU/ml). This may possibly be attributed to preventing the loss of nisin due the detrimental effects and a higher biomass density achieved during online recovery process, which stimulated production of nisin during fermentation.     

Listeriosis in Sheep

Two hundred and ninety-one grass silage samples from 113 farms with recent outbreaks of listeriosis were examined for the presence of Listeria monocytogenes (Lm). The frequency of Lm isolations increased with increasing pH. Lm was isolated from 22 % of the samples with pH < 4, from 37 % with pH 4–5 and from 56 % with pH > 5. Formic acid had been used as additive. A similar investigation was carried out on 32 samples from a farm with no outbreak of listeriosis during the investigation period. Lm was isolated from 9 samples.     

改善啤酒酿造过程酶活力方法的研究

以国外大麦Gairdner为原料,分剐用麦芽厂所用的空白水、pH值4．0盐酸溶液、pH值4．0硫酸溶液、100mg／L钙离子溶液、1μg／L赤霉素溶液使大麦浸渍、发芽来达到降低绿麦芽内部pH值的目的,实验结果表明．虽然硫酸、盐酸2种强酸物质具备降低大麦发芽内部pH值的能力,但其食品安全性差、对设备腐蚀大,而用钙离子和赤霉素溶液培养大麦发芽,虽然在大麦发芽过程中对pH值的变化影响不是很稳定,但均能使大麦发芽结束点即绿麦芽的pH值降低,所以对于后期啤酒的酿造均能够使淀粉酶、蛋白酶等活力近于最适pH值状态,为在啤酒酿造过程中减少外源添加酸的使用量,或为温和型弱酸或酸性中草药等物质的添加提供条件。     

Characterization of Trametes versicolor laccase for the transformation of aqueous phenol

Laccase (oxygen oxidoreductase, EC 1.10.3.2) from Trametes versicolor was thoroughly characterized in terms of its catalytic stability and its effectiveness as a biocatalyst under various reaction conditions when using phenol as a model substrate. This enzyme demonstrated high or moderate degrees of stability at pHs from 5 to 8 at 25 degrees C and at temperatures from 10 to 30 degrees C at pH 6. Exponential decay expressions were successfully used to model laccase inactivation when incubated under various conditions of pH and temperature. Phenol transformation was optimum at pH 6, but significant transformation was observed over a pH range of 4-7, provided that sufficient laccase was present in the reacting solution. Partial inactivation of laccase was observed during the oxidation of phenol, even under conditions of optimal stability (pH 6 and 25 degrees C).     

Intragastric pH regulates conversion from net acid to net alkaline secretion by the rat stomach

Our previous report showed gastric mucosal surface pH was determined by alkali secretion at intragastric luminal pH 3 but by acid secretion at intragastric pH 5. Here, we question whether regulation of mucosal surface pH is due to the effect of luminal pH on net acid/base secretions of the whole stomach. Anesthetized rats with a gastric cannula were used, the stomach lumen was perfused with weakly buffered saline, and gastric secretion was detected in the gastric effluent with 1) a flow-through pH electrode and 2) a fluorescent pH-sensitive dye (Cl-NERF). During pH 5 luminal perfusion, both pH sensors reported the gastric effluent was acidic (pH 4.79). After perfusion was stopped transiently (stop-flow), net acid accumulation was observed in the effluent when perfusion was restarted (peak change to pH 4.1-4.3). During pH 3 luminal perfusion, both pH sensors reported gastric effluent was close to perfusate pH (3.0-3.1), but net alkali accumulation was detected at both pH sensors after stop-flow (peak pH 3.3). Buffering capacity of gastric effluents was used to calculate net acid/alkaline secretions. Omeprazole blocked acid secretion during pH 5 perfusion and amplified net alkali secretion during pH 3 perfusion. Pentagastrin elicited net acid secretion under both luminal pH conditions, an effect antagonized by somatostatin. We conclude that in the basal condition, the rat stomach was acid secretory at luminal pH 5 but alkaline secretory at luminal pH 3.     

Porous silicon based potentiometric triglyceride biosensor

A novel method for estimating triglycerides is reported. Porous silicon, prepared from p-type (100) crystalline silicon was thermally oxidized and used to immobilise lipase, an enzyme, which hydrolyses triglycerides resulting in the formation of fatty acids. This causes a change in the pH of the solution. Enzyme solution-oxidized porous silicon-crystalline silicon structure was used to detect changes in pH during the hydrolysis of tributyrin as a shift in the capacitance-voltage (C-V) characteristics. Detailed calibration of the sensor is included.     

Effects of ionic speciation of lysine on its adsorption and desorption through a sulfone-type ion-exchange column

Lysine produced during microbial fermentation is usually recovered by an ion-exchange process, in which lysine is first converted to the cationic form (by lowering the pH to less than 2.0 with sulfuric acid) and then fed to a cation-exchange column containing an exchanger that has a sulfone group with a weak counterion such as NH4+. Ammonia water with a pH above 11 is then supplied to the column to displace the purified lysine from the column and allow its recovery. To enhance the adsorption capacity and for a possible reduction in chemical consumption, monovalent lysine fed at pH 4 was investigated in comparison with conventional divalent lysine fed at pH 1.5. The adsorption capacity increased by more than 70% on a mass basis using pH 4 feeding compared with pH 1.5 feeding. Lysine adsorbed at pH 4 started to elute earlier than that adsorbed at pH 1.5 when ammonia water was used as the eluant solution, and the extent of early elution became more notable at lower concentrations of ammonia. Moreover, the elution of monovalent lysine fed at pH 4 displayed a stiffer front boundary and higher peak concentration. However, when the ammonium concentration was greater than 2.0 N, complete saturation of the bed was delayed during adsorption and the percent recovery yield from elution was lowered, both drawbacks that were considered inevitable features originating from the increased adsorption of monovalent lysine.     

Hydrogen production from sewage sludge using mixed microflora inoculum: effect of pH and enzymatic pretreatment

Hydrogen was successfully produced by fermenting primary sewage sludge which had been both heat treated and digested with a commercially available enzyme preparation. When either heat treatment or enzymatic digestion were not used, no hydrogen was produced during fermentation. Heat treated mesophilic anaerobic sludge was used as an inoculum rather than a pure microbial culture. Fermentation was conducted at pH levels ranging from of 4.5 to 7.0. When fermentation took place at pH 5.5 a peak hydrogen production rate of 3.75 ml min(-1) was observed. At this pH the hydrogen yield was 0.37 mol H(2)mol(-1) carbohydrate, equivalent to 18.14L H(2)kg(-1) dry solids.     

Effect of hydraulic retention time on anaerobic hydrogenesis in CSTR

The objective of this work was to evaluate the production of hydrogen in a continuous system as a function of hydraulic retention time (HRT). The intermediates accumulated and other parameters of pH, oxidation-reduction potential were quantified. The heat treatment (103 degrees C for 24 h) of the compost from a cattle dung composting facility was able to select H2-producing spores; this product was used as a seed for continuous systems. The brewery waste was used as substrate. For the eight runs with combinations of five HRTs and four pHs, the results indicate that at pH=5.5, a maximum H2 production of 47% H2 concentration, 43 ml H2/g COD(added), and 3.1 l H2/l reactor d was achieved at HRT=18 h. Nevertheless, at HRT=18 h, pH 5.5 was also the optimum pH for the maximum H2 production among four pHs evaluated from 5 to 6.5. There was a significant accumulation of volatile acid and alcohols during the entire study.     

The effect of chemical crosslinking of invertase with dimethyl suberimidate on its pH stability

The effect of chemical crosslinking of invertase with a homo-bifunctional bisimidoester on its pH stability was studied. Dimethyl suberimidate (DMS) was used as the crosslinker and pH inactivation was investigated in the pH range of 2.5–10. The inactivation mechanisms of both native and DMS crosslinked invertases were observed to follow first-order kinetics during prolonged incubation. Although DMS crosslinking of invertase increased the pH stability of the enzyme over the complete pH range, it was especially effective over the acidic range. The half-lives of invertase increased almost twofold, on average, by crosslinking at the neutral to the acidic range. The effect of crosslinking was especially pronounced at pH 5 since both the half-life of the native invertase and the stabilization factor were very high. Higher activation free energies of inactivation were obtained for DMS crosslinked invertases over the whole pH range which also indicates the stabilization of invertase by crosslinking.     

Characterization of proteolytic activities in embryos of Xenopus laevis

1. Proteolytic activities in early embryos of Xenopus laevis exhibited maximum levels at pH 3.2, 5.6 and 7.2 when 3H-BSA was used as substrate, and the maximum proteolytic activity at pH 3.2 was several thousand-fold higher during the tail bud stage than in the unfertilized egg. 2. The proteolytic activity at pH 3.2 was separated into two fractions by gel chromatography. One fraction corresponded to a mol. wt of about 40,000 and its activity was inhibited by thiol protease inhibitors. The other appeared to be a protease of much higher mol. wt. 3. The maximum activities at pH 5.6 and 7.2 appear to correspond to proteins of mol. wt greater than 1,000,000.