无机碳源对小球藻自养产油脂的影响

旨在研究小球藻利用无机碳自养产油脂,考察了3种无机碳源 (Na2CO3、NaHCO3和CO2) 及其初始浓度对小球藻产油特性的影响。结果表明,小球藻能利用Na2CO3、NaHCO3和CO2产油;经Na2CO3、NaHCO3和CO2培养10 d后,随着每种无机碳源浓度的增加,小球藻产量均先增加后减少。小球藻经3种无机碳源培养后,其培养液pH值上升。最适宜的Na2CO3和NaHCO3添加量均为40 mmol/L,其生物量分别达到0.52 g/L和0.67 g/L,产油量分别达到0.19 g/L和0.22 g/L。在3种无机碳源中,CO2是最佳无机碳源,当CO2浓度为6％时,小球藻生长最快,生物量达2.42 g/L,产油量最高达0.72 g/L;当CO2浓度过低时,无机碳供应不足,油脂产量低;当CO2浓度过高时,培养液pH偏低,小球藻油脂积累受到抑制。Na2CO3和NaHCO3较CO2更有利于小球藻积累不饱和脂肪酸。     

盐碱土壤与非盐碱土壤微生物的抗盐碱特性

为了比较分析盐碱土壤与非盐碱土壤微生物资源抗盐碱性差异,本研究利用含有不同浓度Na2CO3、NaHCO3和pH的培养基对盐碱土壤和非盐碱土壤细菌进行培养计数.结果显示:非盐碱土壤出菌数量随Na2CO3、pH和NaHCO3浓度升高而下降,盐碱土壤细菌出菌数量随着Na2CO3、pH和NaHCO3浓度升高先是升高然后下降,最高值分别出现在200 mmol/L NaHCO3、50 mmol/L Na2CO3和pH 9.0的分离平板上.此外,高Na2CO3、pH和NaHCO3浓度的平板中盐碱土壤出菌数量远高于非盐碱土壤;以上结果可见,耐盐碱细菌资源主要集中分布在盐碱土壤中,在非盐碱土壤中虽有分布,但是仅占有很少一部分.     

长穗薄冰草（Thinopyrum ponticum）对盐碱胁迫的生理响应

实验采用不同浓度（以干土重0.3%、0.5%、0.7%和0.9%计）Na2SO4、NaCl、NaHCO3、Na2CO3,Na2SO4：NaCl=1：1、NaHCO3：Na2CO3=1：1和Na2SO4：NaCl：NaHCO3：Na2CO3=1：1：1：1对长穗薄冰草进行胁迫,研究长穗薄冰草对盐碱胁迫的生理响应.结果表明：根系活力、SOD活性随盐碱浓度的增加逐渐下降,脯氨酸含量呈上升趋势,叶绿素含量除Na2SO4处理组外,呈先上升后下降的趋势.其中碱性盐（NaHCO3、Na2CO3、NaHCO3：Na2CO3=1：1、Na2SO4：NaCl：NaHCO3：Na2CO3=1：1：1：1）对长穗薄冰草的胁变效应较中性盐（Na2SO4、NaCl、Na2SO4：NaCl=1：1）大,以Na2SO4处理组对长穗薄冰草的影响最小,0.7% Na2CO3是长穗薄冰草正常生长的临界浓度.     

牛叠肚幼苗对盐碱胁迫的生理响应及其耐盐阈值

以盆栽牛叠肚组培苗为试材,比较研究了不同浓度中性盐(NaCl、Na2SO4)和碱性盐(NaHCO3、Na2CO3)胁迫对其生长和生理指标的影响。结果显示:(1)牛叠肚幼苗生长在碱性盐(NaHCO3、Na2CO3)处理下表现出"低促高抑"现象,而在中性盐(NaCl、Na2SO4)处理下均受到不同程度的抑制。(2)随着盐碱胁迫浓度的升高,牛叠肚叶片的相对电导率呈增加趋势,丙二醛(MDA)积累波动变化;Na2SO4和NaHCO3处理下二者之间的变化趋势相似,而NaCl和Na2CO3处理下二者之间变化趋势则不同。(3)牛叠肚叶片中超氧化物歧化酶(SOD)活性随胁迫浓度增加先升高后下降,而过氧化物酶(POD)活性呈先下降后升高趋势,说明牛叠肚主要通过SOD和POD的互补作用来降低氧化伤害。(4)以相对株高生长量下降50%为标准,求得牛叠肚幼苗对NaCl、Na2SO4、NaHCO3、Na2CO34种单盐的耐受阈值分别为85.18(0.50%,W/V)、40.77(0.58%,W/V)、171.00(1.44%,W/V)、114.20(1.21%,W/V)mmol·L-1。研究表明,各盐碱胁迫使牛叠肚幼苗的生长受到不同程度的抑制,但其在一定浓度范围内通过提高抗氧化酶(SOD、POD)活性来减轻盐碱伤害,维持植株的正常生理代谢;牛叠肚幼苗对碱性盐(NaHCO3、Na2CO3)的耐受能力强于中性盐(NaCl、Na2SO4)。     

米根霉发酵产富马酸的最适替代中和剂及pH调控策略研究

针对米根霉发酵产富马酸使用的不同中和剂(CaCO3,Na2CO3,NH3·H2O,NaOH)进行了研究,结果表明发酵过程中使用Na2CO3作为中和剂时富马酸产率和生产强度最接近传统中和剂CaCO3.此后考察了不同pH值(3.5,4.5,5.5和6.5)对Na2CO3作为中和剂的富马酸发酵过程的影响.基于对3个动力学参数的分析,提出了一个旨在同时获得富马酸高产物浓度、高产率和高生产强度的双阶段pH调控策略,在初始的24 h内pH控制在5.5,然后将pH调到4.5直至发酵结束.最终富马酸的终浓度达到40.5 g/L,产率为0.55 g/g,生产强度为0.61 g/L/h,比恒定pH时的最优结果分别提高了8.3％,10.0％和17.3％,其中生产强度甚至比使用CaCO3时还高了3.4％.故以Na2CO3作为中和剂,采用双阶段pH调控策略具有降低能耗和简化下游步骤的优势,可以成功取代CaCO3.     

紫杆柽柳MnSOD基因在非生物胁迫中的作用

构建了酵母表达载体pYES2-MnSOD,并将其转入酿酒酵母INVSc1,以转空载体pYES2的酵母INVSc1为对照,检测其抗NaCl、Na2CO3、NaHCO3和紫外线胁迫能力。结果表明,转MnSOD基因酵母在以上逆境因素胁迫下的存活率有明显提高,说明MnSOD基因具有较强的抗NaCl、Na2CO3、NaHCO3和紫外线胁迫能力。     

盐碱协同胁迫对向日葵抗氧化酶系统的影响

根据中国东北盐碱土壤特点,将4种盐NaCl、NaHCO3、Na2SO4和Na2CO3按不同比例混合,模拟出25种盐度和pH值各不相同的复杂盐碱条件(盐浓度为50～250 mmol/L,pH值为712～1046),并对向日葵苗进行盐碱混合胁迫处理,研究了向日葵超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)酶等抗氧化酶系统和丙二醛(MDA)的盐碱协同胁迫效应．结果表明, 向日葵抗氧化物酶活性强弱同时与盐度和碱度密切相关,3种抗氧化物酶活性对于盐浓度的反应相似,均为其含量随着盐浓度的升高开始逐渐升高然后下降,而对于pH的影响,不同酶反应结果不同．即随着pH值升高,SOD酶活性和CAT酶活性降低,而POD酶活性反应则是随着pH值升高活性也升高．双向方差分析(ANOVA)结果表明：盐碱效应对于3种酶活力的影响是显著的．其中,盐效应对POD和SOD活性的影响比pH值的影响大,而pH值对CAT活性的影响效应比盐效应大．除SOD外,盐碱效应的交互作用显著 (P<0001)．抗氧化酶系统和MDA含量两者间相关性和逐步回归分析表明,3种酶对MDA的影响效应随其强度不同呈现显著不同．其中SOD是1个主导因子,CAT 处于次位, 而POD的影响不大,甚至可以忽略．     

混合盐碱胁迫对芹菜种子萌发的影响

NaCl、Na2SO4、NaHCO3和Na2CO3按不同比例混合,模拟盐度和pH变化规律与天然盐碱地相似的15种盐碱条件,探讨混合盐碱胁迫对芹菜(Apium graveolens)种子萌发的影响.结果表明:芹菜种子的发芽率、发芽势、发芽指数和活力指数均随盐浓度的升高,pH的增大呈下降趋势.芹菜种子的萌发主要受盐浓度的影响,不同盐浓度间的影响差异大;当盐浓度为200 mmol/L时,基本不萌发.     

NaHS对NaHCO_3胁迫下黑籽南瓜种子萌发及生理特性的影响

以黑籽南瓜(Cucurbita ficifolia)种子为试材,研究了外施不同浓度的NaHS对NaHCO3胁迫下种子萌发及生理特性的影响。结果表明,NaHCO3胁迫显著抑制了黑籽南瓜种子的发芽率、胚轴长和胚根长,降低了种子萌发过程中的可溶性糖含量,抑制了α-淀粉酶、β-淀粉酶、SOD及POD活性。而外施不同浓度的NaHS显著促进了NaHCO3胁迫下黑籽南瓜萌发种子胚轴和胚根的生长,提高了可溶性糖含量及α-淀粉酶、β-淀粉酶、SOD和POD活性,降低了MDA含量;外施其它盐类(Na2S、Na2SO4、NaHSO4和NaHSO3)及不同pH值(pH5.8–7.8)的Na2HPO4-NaH2PO4缓冲液对NaHCO3胁迫下黑籽南瓜种子的萌发则无影响。外施NaHS可有效缓解NaHCO3胁迫对黑籽南瓜种子萌发的抑制作用,其缓解效应可能与其释放的H2S有关。     

Fuchsin staining with naoh clearing for lignified elements of whole plants or plants organs

Three methods that are adapted to the various consistencies of plants are as follows: 1. Samples are placed for 10-14 hr at 60° C in a 1% aqueous solution of basic fuchsin, to which 10 gm of solid NaOH per 100 ml are added. 2. Samples when taken out of 95% alcohol are placed in a 1% solution of basic fuchsin in 95% alcohol for 24 hr; after washing in water, they are placed in a 15% solution of NaOH at 60° C until cleared. 3. Samples are placed in a 15% aqueous solution of NaOH at 60° C until cleared, then for 24 hr at 60° C in 15% NaOH containing basic fuchsin. After being stained and cleared by one of these three methods, the samples are rinsed in water, dehydrated and then passed into a mixture of absolute alcohol and concentrated HC1 (3:1) for 1-15 min, rinsed in absolute alcohol, cleared in xylene and mounted in Canada balsam. The lignified tissues appear red; the others, transparent.     

Determination of aconitine,hypaconitine and mesaconitine in urine using hollow fiber liquid-phase microextraction combined with high-performance liquid chromatography

Hollow fiber liquid-phase microextraction (HF-LPME) coupled with high-performance liquid chromatography was used to simultaneously determine three Aconitum alkaloids, including aconitine (AC), hypaconitine (HA) and mesaconitine (MA) in human urine sample. Analytes were extracted from 5 mL urine sample containing 1.0 mmol/L NaOH into 1-octanol membrane phase impregnated in the pores of hollow fiber wall, and then back extracted into acidified aqueous solution in the lumen of the hollow fiber. After extraction, 10 μL of the acceptor phase was analyzed directly by HPLC. In this method, some important extraction parameters, such as organic solvent, extraction time, stirring rate, pH of donor phase and acceptor phase, temperature, and the volume of acceptor phase were optimized. This method provided 98- to 288-fold enrichment factors within 60 min of extraction and good repeatability with RSDs of 0.99–7.22%. The calibration curves were linear over the ranges of 16.0–128.0 μg/L for AC, 11.0–88.0 μg/L for HA and 8.1–64.8 μg/L for MA in human urine sample, with correlation coefficients of 0.9949, 0.9969 and 0.9904, respectively. Limits of detection were from 0.7 to 1.5 μg/L, and recoveries from spiked urine sample varied from 84.4% to 106.2% for AC, 77.3% to 85.6% for HA and 90.1% to 100.8% for MA.     

Acute renal response to rapid onset respiratory acidosis

Renal strong ion compensation to chronic respiratory acidosis has been established, but the nature of the response to acute respiratory acidosis is not well defined. We hypothesized that the response to acute respiratory acidosis in sheep is a rapid increase in the difference in renal fractional excretions of chloride and sodium (Fe(Cl) - Fe(Na)). Inspired CO(2) concentrations were increased for 1 h to significantly alter P(a)CO(2) and pH(a) from 32 ± 1 mm Hg and 7.52 ± 0.02 to 74 ± 2 mm Hg and 7.22 ± 0.02, respectively. Fe(Cl) - Fe(Na) increased significantly from 0.372 ± 0.206 to 1.240 ± 0.217% and returned to baseline at 2 h when P(a)CO(2) and pH(a) were 37 ± 0.6 mm Hg and 7.49 ± 0.01, respectively. Arterial pH and Fe(Cl) - Fe(Na) were significantly correlated. We conclude that the kidney responds rapidly to acute respiratory acidosis, within 30 min of onset, by differential reabsorption of sodium and chloride.     

碳源对粉核油球藻生长和脂肪酸组成特性的影响

研究了不同碳源类型（CO2、NaHCO3和葡萄糖）及其浓度对粉核油球藻（Pinguiococcus pyrenoidosus CCMP 2078）生长及脂肪酸组成的影响。结果表明：（1）培养液中适量添加碳源促进了粉核油球藻的生长,三种碳源的适宜添加浓度分别是0.5% CO2,5mmol/L NaHCO3和20g/L葡萄糖,对数生长末期的细胞密度分别是对照的3.10倍、1.47倍和2.78倍;（2）除了低浓度葡萄糖外,其他碳源类型和浓度均降低了TPUFA和EPA占总脂肪酸的比例,提高了TSFA的比例,胞内EPA和TSFA含量均下降;（3）低浓度碳源提高了TSFA和EPA产量。通入0.5% CO2培养的EPA和TSFA产量分别是对照的2.30倍和2.69倍,5mmol/L NaHCO3培养的TSFA产量是对照的1.85倍,5g/L和10g/L葡萄糖培养的EPA和TSFA产量最高分别可达对照的2.11倍和1.58倍。因此,通入低浓度CO2最有利于粉核油球藻的生长以及EPA和饱和脂肪酸的生产,EPA和饱和脂肪酸含量的提高主要是通过生物量的增大来实现的。     

Rat kidney mitochondrial carbonic anhydrase

Mitochondrial carbonic anhydrase has previously been quantitated in liver mitochondria; it was not detected in guinea pig kidney cortical mitochondria. Evidence of this enzyme in rat kidney cortical mitochondria is reported. Electron microscopy showed that intact mitochondria were free of other intracellular organelles. When intact kidney mitochondria were added to isotonic 3'-(N'-morpholino) propanesulfonic acid buffer with 25 mM KHCO3 (1% labeled with 18O) the rate of disappearance of C18O16O was biphasic; this indicates that there is carbonic anhydrase within the inner mitochondrial membrane. Intact rat kidney mitochondria were assayed for carbonic anhydrase activity at 4 degrees C by the changing pH technique. The rate of CO2 hydration in the presence and absence of intact mitochondria was identical; this rate increased when Triton X-100 was added which indicates that all carbonic anhydrase is inside the inner mitochondrial membrane. Carbonic anhydrase activity was quantitated as kenz (units, ml.s-1 mg-1 mitochondrial protein) at 37 degrees C, pH 7.4, in 25 mM NaHCO3 (1% labeled with 18O) by following the rate of disappearance of C18O16O from solutions before and after addition of disrupted mitochondria. Values of Kenz for liver and kidney mitochondria from rats given free access to normal rat chow and water at neutral pH were 0.06 and 0.08 (respectively). Values of kenz for liver and kidney mitochondria from rats fed as above and with free access to water adjusted to pH 2.5 with HCl were 0.04 and 0.16, respectively. Values of kenz for rats starved for 48 h were 0.06 and 0.12 (respectively). The values of kenz remained 0.11-0.14 in liver mitochondria from guinea pigs fed normally, given dilute acid, or starved and the value was always at zero in guinea pig kidney mitochondria. Values of Kenz were measured with disrupted mitochondria by the 18O technique as a function of pH at 25 degrees C, 25 to 75 mM NaHCO3, ionic strength 0.3. From pH 7.0 to 8.0 kenz increased threefold for mitochondria from rat liver, fed rat kidney, and acid rat kidney, and increased eightfold for mitochondria from guinea pig liver. kenz was decreased similarly by increasing HCO3- in mitochondria from rat liver, fed kidney, and acid kidney; it is concluded that carbonic anhydrase in rat liver mitochondria is probably the same isozyme as in rat kidney mitochondria. The published observation that rat kidney cortices are up to 10 times as gluconeogenic from pyruvate as guinea pig kidney cortices can be explained by the presence of mitochondrial carbonic anhydrase in rat but not guinea pig mitochondria.     

Factors modulating the blood feeding behavior and the electrophysiological responses of labral apical chemoreceptors to adenine nucleotides in the mosquito Aedes aegypti (Culicidae)

The feeding of Aedes aegypti (L.) on blood is induced by the presence of phagostimulants: adenine nucleotides. Three chemoreceptive cells in the labral apical sensilla can distinguish the presence of adenine nucleotides depending on the other stimulus components. This work aims at correlating the sensory information arising from the labral apical sensilla with the feeding behavior in response to the same stimuli. The saline stimulating solution, containing adenine nucleotides, is modulated by changing one of the following components: salt concentration, buffer or pH. Cell 3 that responds to NaCl in a dose dependent manner seems to have another unique modality. The response of this cell is unaffected by ATP when the stimulating solution is NaCl buffered by NaHCO(3). It responds at a higher spike frequency to the presence of ATP in a NaCl solution without NaHCO(3). Thus in the presence of ATP Cell 3 detects whether the NaCl solution is buffered by NaHCO(3). Both the blood feeding response and the sensory information from Cell 2 (which responds at high spike frequencies to the presence of ATP) are modulated by pH in a similar way. Both responses present a bi-modal response, with a major peak at pH 4.0 and a moderate peak at the most alkaline pH value tested.     

Micelles of cerebroside sulphate

The critical micelle concentration of cerebroside sulphate in water is 0-01 mM: it increases with increasing concentrations of buffer to 0-07 mM in 0-1 M sodium acetate and formate buffers, pH 5-6 and 4-5 respectively. The partial specific volume of the micelles is about 0-94. The behaviour of the micelles in the ultracentrifuge and on Sephadex G-200 shows them to be grossly heterogeneous with respect to size. In 0-1 M buffer s20,w is about 26 S; in water or 0-01 M buffer smaller micelles with an s20,w of about 6 S are also present. In 0-01 M formate, pH 4-5, the smallest species detectable by equilibrium ultracentrifugation had a micellar weight of about 180,000 corresponding to an aggregation number of about 180. Much larger aggregates were also present. It is suggested that the smallest micelles are the substrate for sulphatase A when this is acting as a cerebroside sulphatase in buffers of low ionic strength.     

Kinetics of enzymatic solid-to-solid peptide synthesis: synthesis of Z-aspartame and control of acid-base conditions by using inorganic salts

Enzymatic peptide synthesis can be carried out efficiently in solid-to-solid reaction mixtures with 10% (w/w) water added to a mixture of substrates. The final reaction mass contains >/=80% (by weight) of product. This article deals with acid-base effects in such reaction mixtures and the consequences for the enzyme. In the Thermoase-catalyzed synthesis of Z-Asp-Phe-OMe, the reaction rate is strongly dependent on the amount of basic salts added to the system. The rate increases 20 times, as the KHCO(3) or K(2)CO(3) added is raised 2.25-fold from an amount equimolar to the Phe-OMe. HCL starting material. With further increases in KHCO(3) addition, the initial rate remains at the maximum, but with K(2)CO(3) it drops sharply. Addition of NaHCO(3) is less effective, but rates are faster if more water is used. With >1.5 equivalents of basic salt, the final yield of the reaction decreases. Similar effects are observed when thermolysin catalyzes the same reaction, or Z-Gln-Leu-NH(2) synthesis. These effects can be rationalized using a model estimating the pH of these systems, taking into account the possible formation of up to ten different solid phases.     

产油微藻的分离、筛选及自养培养氮源、碳源的优化

从云南滇池的水样中分离筛选得到一株自养产油小球藻(Chlorella vulgaris,C.vulgaris),其油脂产率可达28.6mg/(L·d),进一步考察了不同氮源、氮源浓度和添加无机碳源对其自养生长和油脂积累的影响。结果表明,硝酸钠为优化氮源,氮元素的优化浓度为123mg/L,油脂含量随氮元素浓度升高而降低;添加NaHCO3显著提高了C.vulgaris生物量产率和油脂产率,其优化浓度为800mg/L。在氮源和碳源的优化浓度下,C.vulgaris的最大生物量产率和油脂产率可达332.8mg/(L·d)和100.2mg/(L·d),分别是对照组的3.6和3.4倍。     

Evaluation of clastogenicity of formic acid, acetic acid and lactic acid on cultured mammalian cells

Using Chinese hamster ovary K1 cells, chromosomal aberration tests were carried out with formic acid, acetic acid and lactic acid, and the relationship between the pH of the medium and the clastogenic activity was examined. The medium used was Ham's F12 supplemented with 17 mM NaHCO3 and 10% fetal calf serum. All of these acids induced chromosomal aberrations at the initial pH of ca. 6.0 or below (about 10-14 mM of each acid) both with and without S9 mix. Exposure of cells to about pH 5.7 or below (about 12-16 mM of each acid) was found to be toxic. When the culture medium was first acidified with each of these acids and then neutralized to pH 6.4 or pH 7.2 with NaOH, no clastogenic activity was observed. Using F12 medium supplemented with 34 mM NaHCO3 as a buffer, no clastogenic activity was observed at doses up to 25 mM of these acids (initial pH 5.8-6.0). However, it was found that about 10% of the cells had aberrations at pH 5.7 or below (27.5-32.5 mM of each acid). Furthermore, when 30 mM HEPES was used as a buffer, chromosomal aberrations were not induced at doses up to 20 mM formic acid and acetic acid (initial pH 7.0-7.1), and at doses up to 30 mM lactic acid (initial pH 6.6). In the initial pH range of 6.4-6.7 (25-32.5 mM of each acid), chromosomal aberrations were observed. The above results show that these acids themselves are non-clastogenic, and the pseudo-positive reactions attributable to non-physiological pH could be eliminated by either neutralization of the treatment medium or enhancement of the buffering ability.