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排序方式: 共有169条查询结果,搜索用时 15 毫秒
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Nikel PI Pettinari MJ Galvagno MA Méndez BS 《Applied microbiology and biotechnology》2008,77(6):1337-1343
Poly(3-hydroxybutyrate) (PHB) synthesis was analyzed under microaerobic conditions in a recombinant Escherichia coli arcA mutant using glycerol as the main carbon source. The effect of several additives was assessed in a semi-synthetic medium by the 'one-factor-at-a-time' technique. Casein amino acids (CAS) concentration was an important factor influencing both growth and PHB accumulation. Three factors exerting a statistically significant influence on PHB synthesis were selected by using a Plackett-Burman screening design [glycerol, CAS, and initial cell dry weight (CDW) concentrations] and then optimized through a Box-Wilson design. Under such optimized conditions (22.02 g l(-1) glycerol, 1.78 g l(-1) CAS, and 1.83 g l(-1) inoculum) microaerobic batch cultures gave rise to 8.37 g l(-1) CDW and 3.52 g l(-1) PHB in 48 h (PHB content of 42%) in a benchtop bioreactor. Further improvements in microaerobic PHB accumulation were obtained in fed-batch cultures, in which glycerol was added to maintain its concentration above 5 g l(-1). After 60 h, CDW and PHB concentration reached 21.17 and 10.81 g l(-1), respectively, which results in a PHB content of 51%. Microaerobic fed-batch cultures allowed a 2.57-fold increase in volumetric productivity when compared with batch cultures. 相似文献
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新乡市水源地土壤重金属含量特征及其对地下水的影响 总被引:3,自引:0,他引:3
对新乡市自来水水源地土壤和地下水重金属含量特征进行分析,结果表明:土壤中Pb含量达标,Zn、Cr、Ni和Cu含量超标,其平均含量分别为1213.80、115.22、189.56和60.39mg.kg-1;用化学连续浸提法对超标元素化学形态分布进行研究表明,Zn、Cr、Ni和Cu残渣态含量最高,所占比例分别为87.81%、78.38%、84.26%和59.33%,可交换态含量最低,所占比例分别为0.21%、2.56%、1.00%和3.51%,由于Zn、Cr、Ni、Cu主要以迁移性最弱的残渣态为主,未发现地下水中重金属含量超标。 相似文献
45.
Manuel S. Godoy Pablo I. Nikel José G. Cabrera Gomez M. Julia Pettinari 《Applied and environmental microbiology》2016,82(1):244-254
The CreBC (carbon source-responsive) two-component regulation system of Escherichia coli affects a number of functions, including intermediary carbon catabolism. The impacts of different creC mutations (a ΔcreC mutant and a mutant carrying the constitutive creC510 allele) on bacterial physiology were analyzed in glucose cultures under three oxygen availability conditions. Differences in the amounts of extracellular metabolites produced were observed in the null mutant compared to the wild-type strain and the mutant carrying creC510 and shown to be affected by oxygen availability. The ΔcreC strain secreted more formate, succinate, and acetate but less lactate under low aeration. These metabolic changes were associated with differences in AckA and LdhA activities, both of which were affected by CreC. Measurement of the NAD(P)H/NAD(P)+ ratios showed that the creC510 strain had a more reduced intracellular redox state, while the opposite was observed for the ΔcreC mutant, particularly under intermediate oxygen availability conditions, indicating that CreC affects redox balance. The null mutant formed more succinate than the wild-type strain under both low aeration and no aeration. Overexpression of the genes encoding phosphoenolpyruvate carboxylase from E. coli and a NADH-forming formate dehydrogenase from Candida boidinii in the ΔcreC mutant further increased the yield of succinate on glucose. Interestingly, the elimination of ackA and adhE did not significantly improve the production of succinate. The diverse metabolic effects of this regulator on the central biochemical network of E. coli make it a good candidate for metabolic-engineering manipulations to enhance the formation of bioproducts, such as succinate. 相似文献
46.
目的:探索建立老年人跌倒防控信息管理系统,并进行应用及效果评价。方法:依托物联网技术和信息化手段,将老年人跌倒综合防控策略从线下导入线上,建立集风险评估、远程教育、反馈评价于一体的老年人跌倒防控信息管理系统。运用该系统对126名干休所老年人(年龄≥ 60岁)进行评估,量化跌倒风险等级,对筛选出的84名高危老年人进行远程连续综合干预,干预措施包括分发宣传画册,制作宣教幻灯片及视频进行远程播放与讲解。采用自身前后对照,分析评价干预6个月后跌倒相关情况。结果:高危人群跌倒发生率由21.43%降至4.76%(P<0.01);身体平衡能力和步态稳定性改善情况明显(P<0.01);能够自觉进行正确防控行为的比例显著提高(P<0.01);进行复杂行为时更有信心不会跌倒(P<0.01);居家环境安全性整体提高(P<0.01)。结论:老年人跌倒防控信息管理系统可通过全面评估筛选出高危人群,实施远程连续综合干预,降低跌倒发生率,可进一步推广应用。 相似文献
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三种不同泌氧能力的红树植物对铅、锌、铜的耐性研究 总被引:1,自引:0,他引:1
红树林是分布于热带、亚热带潮间带的典型滨海生态系统。近年来,随着城市化和工业化的发展,滨海生态系统受重金属(例如铅、锌、铜等)的污染越来越严重,我国南方一些红树林底泥中的重金属浓度已达到甚或超过重度污染标准。由于沉积作用,红树林底泥被认为是一个能积累由潮水和河流淡水携带来的重金属的"库"。然而,红树植物具有很强的耐性生长于重度污染的底泥, 但其中的机理目前尚不清楚。由于长期被水淹没,红树林底泥是一个具氧化还原势低,还原性毒性物质(如Fe2+、Mn2+、H2S、CH4等)积累多,营养物质缺乏等特征的厌氧环境。为了适应这种生境,红树植物也进化出了一套与其他湿地植物类似的适应机制,植物能通过通气组织将地上部分的氧气输送到地下,一部分满足根的呼吸作用需要,一部分则通过根释放到根际,这被称为根的泌氧。根的泌氧可以氧化根际环境并且氧化还原性的毒性物质,以保证红树植物根免受毒害而延长生长。因此,根际泌氧是红树植物能适应生境的一个重要机理。本研究试图揭示红树植物在长期适应生境的过程中已经进化和发展出一系列形态解剖和生理生化特征是否在耐浸水的土壤条件,解除Fe2+、Mn2+等元素的毒性同时,是否也能解除其他重金属(铅、锌、铜)的毒性。通过对木榄、桐花和白骨壤三种不同泌氧能力的植物进行8周室内砂培试验发现:(1)三种植物的生长都被铅、锌、铜所抑制;(2)三种植物对铅、锌、铜都具有一定的耐性,然而,泌氧能力较弱的木榄比泌氧能力较强的桐花和白骨壤对铅、锌、铜的耐性高。 相似文献
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50.
Pablo I. Nikel Andrea M. Giordano Alejandra de Almeida Manuel S. Godoy M. Julia Pettinari 《Applied and environmental microbiology》2010,76(22):7400-7406
The effect of eliminating d-lactate synthesis in poly(3-hydroxybutyrate) (PHB)-accumulating recombinant Escherichia coli (K24K) was analyzed using glycerol as a substrate. K24KL, an ldhA derivative, produced more biomass and had altered carbon partitioning among the metabolic products, probably due to the increased availability of carbon precursors and reducing power. This resulted in a significant increase of PHB and ethanol synthesis and a decrease in acetate production. Cofactor measurements revealed that cultures of K24K and K24KL had a high intracellular NADPH content and that the NADPH/NADP+ ratio was higher than the NADH/NAD+ ratio. The ldhA mutation affected cofactor distribution, resulting in a more reduced intracellular state, mainly due to a further increase in NADPH/NADP+. In 60-h fed-batch cultures, K24KL reached 41.9 g·liter−1 biomass and accumulated PHB up to 63% ± 1% (wt/wt), with a PHB yield on glycerol of 0.41 ± 0.03 g·g−1, the highest reported using this substrate.Poly(3-hydroxybutyrate) (PHB) is the best-known and most common polyhydroxyalkanoate (PHA). PHAs are polymers with thermoplastic properties that are totally biodegradable by microorganisms present in most environments and that can be produced from different renewable carbon sources (38). Accumulated as intracellular granules by many bacteria under unfavorable conditions (1, 21), PHAs are carbon and energy reserves and also act as electron sinks, enhancing the fitness and stress resistance of bacteria and contributing to redox balance (12, 30). Escherichia coli offers a well-defined physiological environment for the construction and manipulation of various metabolic pathways to produce different bioproducts, such as PHB, from cost-effective carbon sources.In recent years, a significant increase in the production of biodiesel has caused a sharp fall in the cost of glycerol, the main by-product of biodiesel synthesis. As a result, glycerol has become a very attractive substrate for bacterial fermentations (10), specially for reduced products, such as PHB (36). The E. coli strain used in this work, K24K, carries phaBAC, the structural genes responsible for PHB synthesis, from Azotobacter sp. strain FA8 (23) (Table (Table1).1). The pha genes in K24K are expressed from a chimeric promoter and consequently are not subject to the genetic regulatory systems present in natural PHA producers. Because of this, it can be assumed that regulation of PHA synthesis in the recombinants is restricted by enzyme activity levels, modulated principally by substrate availability. In most natural producers, and also in PHB-producing E. coli recombinants, PHB is synthesized through the condensation of two molecules of acetyl-coenzyme A (acetyl-CoA), catalyzed by an acetoacetyl-CoA transferase or 3-ketothiolase, resulting in acetoacetyl-CoA. This compound is subsequently reduced by an NAD(P)H-dependent acetoacetyl-CoA reductase to R-(−)-3-hydroxybutyryl-CoA, which is then polymerized by a specific PHA synthase (34).
Open in a separate windowaStrain obtained through the E. coli Genetic Stock Center, Yale University, New Haven, CT.bFor oligonucleotides, the ATG codon of ldhA is underlined and the sequences with homology to FRT-kan-FRT in the template plasmid pKD4 (11) are shown in boldface.Cells growing on glycerol are in a more reduced intracellular state than cells grown on glucose under similar conditions of oxygen availability. This has a significant effect on the intracellular redox state, which causes the cells to direct carbon flow toward the synthesis of more-reduced products when glycerol is used than when glucose is used in order to achieve redox balance (31). When metabolic product distribution was analyzed in bioreactor cultures of K24K using glucose or glycerol as the substrate, product distributions with the two substrates were found to be different, as glycerol-grown cultures produced smaller amounts of acetate, lactate, and formate and more ethanol than those grown on glucose. However, PHB production from glycerol was lower than that from glucose, except under conditions of low oxygen availability (13).Manipulations to enhance the synthesis of a metabolic product include several approaches to increase the availability of the substrates needed for its formation or to inhibit competing pathways. The effect of eliminating competing pathways on PHB production from glucose has been investigated through the inactivation of different genes, such as those encoding enzymes participating in the synthesis of acetate (ackA, pta, and poxB) or d-lactate (ldhA). A pta mutant, which produces very little acetate (6), and an frdA ldhA double mutant (40) had increased PHB accumulation from glucose. A recent report using an ackA pta poxB ldhA adhE mutant under microaerobic conditions attained similar results (17). The inactivation of ldhA has also been shown to have an important effect on the metabolic product distribution in recombinant E. coli with glycerol as the carbon source, promoting ethanol synthesis (28). In the present work we analyzed the effect of ldhA inactivation in strain K24K using glycerol as the carbon source, with special emphasis on changes in carbon distribution and in the intracellular redox state, determined through cofactor levels. 相似文献
TABLE 1.
E. coli strains, plasmids, and oligonucleotides used in this studyStrain, plasmid, or oligonucleotide | Relevant characteristicsb | Reference or source |
---|---|---|
E. coli strains | ||
K1060a | F−fadE62 lacI60 tyrT58(AS) fabB5 mel-1 | 29 |
K24 | Same as K1060, carrying pJP24; Apr | 23 |
K24K | Same as K1060, carrying pJP24K; Apr Kmr | 23 |
ALS786a | F− λ−rph-1 ΔldhA::kan; Kmr | 14 |
K24LT | Same as K1060 but ΔldhA::kan by K1060 × P1(ALS786), carrying pJP24; Apr Kmr | This work |
K24KL | Same as K1060 but ΔldhA by allelic replacement, carrying pJP24K; Kmr | This work |
TA3522a | F− λ− Δ(his-gnd)861 hisJo-701 | 2 |
TA3514a | Same as TA3522 but pta-200 | 19 |
TA3522L | Same as TA3522 but ΔldhA::kan by TA3522 × P1(ALS786); Kmr | This work |
TA3514L | Same as TA3514 but ΔldhA::kan by TA3514 × P1(ALS786); Kmr | This work |
Plasmids | ||
pQE32 | Expression vector, ColE1 ori; Apr | Qiagen GmbH, Hilden, Germany |
pJP24 | pQE32 derivative expressing a 4.3-kb BamHI-HindIII insert containing the phaBAC genes from Azotobacter sp. strain FA8 under the control of a T5 promoter/lac operator element; Apr | 23 |
pJP24K | pJP24 derivative; Apr Kmr | 23 |
pCP20 | Helper plasmid used for kan excision; Saccharomyces cerevisiae FLP λ cI857 λ PRrepA(Ts); Apr Cmr | 7 |
Oligonucleotides | ||
ΔldhA-F | 5′-TAT TTT TAG TAG CTT AAA TGT GAT TCA ACA TCA CTG GAG AAA GTC TTA TGG TGT AGG CTG GAG CTG CTT C-3′ | This work |
ΔldhA-R | 5′-CTC CCC TGG AAT GCA GGG GAG CGG CAA GAT TAA ACC AGT TCG TTC GGG CAC ATA TGA ATA TCC TCC TTA G-3′ | This work |