共查询到16条相似文献，搜索用时 0 毫秒
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Robert Huber Marco Scheidle Barbara Dittrich Doris Klee Jochen Büchs 《Biotechnology and bioengineering》2009,103(6):1095-1102
An often underestimated problem when working with different clones in microtiter plates and shake flask screenings is the non‐parallel and non‐equal growth of batch cultures. These growth differences are caused by variances of individual clones regarding initial biomass concentration, lag‐phase or specific growth rate. Problems arising from unequal growth kinetics are different induction points in expression studies or uneven cultivation periods at the time of harvest. Screening for the best producing clones of a library under comparable conditions is thus often impractical or even impossible. A new approach to circumvent the problem of unequal growth kinetics of main cultures is the application of fed‐batch mode in precultures in microtiter plates and shake flasks. Fed‐batch operation in precultures is realized through a slow‐release system for glucose. After differently growing cultures turn to glucose‐limited growth, they all consume the same amount of glucose due to the fixed feed profile of glucose provided by the slow‐release system. This leads to equalized growth. Inherent advantages of this method are that it is easy to use and requires no additional equipment like pumps. This new technique for growth equalization in high‐throughput cultivations is simulated and verified experimentally. The growth of distinctly inoculated precultures in microtiter plates and shake flasks could be equalized for different microorganisms such as Escherichia coli and Hansenula polymorpha. Biotechnol. Bioeng. 2009;103: 1095–1102. © 2009 Wiley Periodicals, Inc. 相似文献
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Perovskite solar cells have delivered power conversion efficiency beyond 22% in less than seven years, implying the potential for the paradigm shift of low‐cost photovoltaics with high efficiency and low embedded energy. Besides the “perovskite fever,” the development of new hole transport materials (HTM), especially dopant‐free HTMs, is another research hotspot. This is because the currently used HTMs, such as spiro‐OMeTAD derivatives, require additional chemical doping process to ensure sufficient conductivity and proper ionic potential level for efficient hole transport and collection. However, the commonly used dopants are volatile and hygroscopic which not only increase the complexity and cost of device fabrication but also deteriorate the device stability. So far, there have been several reviews on new HTMs, but review or analysis on dopant‐free HTMs is scarce. In this review, all reported dopant‐free HTMs are categorized into four primary different types and lessons will be learned during the separate discussions. The stability test behavior of all the intrinsic HTMs will be evaluated directly. In the end, the correlations between the properties of the intrinsic HTMs and parameters of the devices will be plotted to shed light on the future direction of development of this field. 相似文献
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Jakob Michael Müller Simon Bruhn Erwin Flaschel Karl Friehs Joe Max Risse 《Biotechnology progress》2016,32(4):855-864
Streptavidin is a homotetrameric protein binding the vitamin biotin and peptide analogues with an extremely high affinity, which leads to a large variety of applications. The biotin‐auxotrophic yeast Pichia pastoris has recently been identified as a suitable host for the expression of the streptavidin gene, allowing both high product concentrations and productivities. However, so far only methanol‐based expression systems have been applied, bringing about increased oxygen demand, strong heat evolution and high requirements for process safety, causing increased cost. Moreover, common methanol‐based processes lead to large proportions of biotin‐blocked binding sites of streptavidin due to biotin‐supplemented media. Targeting these problems, this paper provides strategies for the methanol‐free production of highly bioactive core streptavidin by P. pastoris under control of the constitutive GAP promoter. Complex were superior to synthetic production media regarding the proportion of biotin‐blocked streptavidin. The optimized, easily scalable fed‐batch process led to a tetrameric product concentration of up to 4.16 ± 0.11 µM of biotin‐free streptavidin and a productivity of 57.8 nM h?1 based on constant glucose feeding and a successive shift of temperature and pH throughout the cultivation, surpassing the concentration in un‐optimized conditions by a factor of 3.4. Parameter estimation indicates that the optimized conditions caused a strongly increased accumulation of product at diminishing specific growth rates (μ ≈ D < 0.01 h?1), supporting the strategy of feeding. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:855–864, 2016 相似文献
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Zhongmin Zhou Xing Li Molang Cai Fengxian Xie Yongzhen Wu Zhang Lan Xudong Yang Yinghuai Qiang Ashraful Islam Liyuan Han 《Liver Transplantation》2017,7(22)
Low‐temperature‐processed perovskite solar cells (PSCs), which can be fabricated on rigid or flexible substrates, are attracting increasing attention because they have a wide range of potential applications. In this study, the stability of reduced graphene oxide and the ability of a poly(triarylamine) underlayer to improve the quality of overlying perovskite films to construct hole‐transport bilayer by means of a low‐temperature method are taken advantage of. The bilayer is used in both flexible and rigid inverted planar PSCs with the following configuration: substrate/indium tin oxide/reduced graphene oxide/polytriarylamine/CH3NH3PbI3/PCBM/bathocuproine/Ag (PCBM = [6,6]‐phenyl‐C61‐butyric acid methyl ester). The flexible and rigid PSCs show power conversion efficiencies of 15.7 and 17.2%, respectively, for the aperture area of 1.02 cm2. Moreover, the PSC based the bilayer shows outstanding light‐soaking stability, retaining ≈90% of its original efficiency after continuous illumination for 500 h at 100 mW cm?2. 相似文献
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Zavrel M Schmidt T Michalik C Ansorge-Schumacher M Marquardt W Büchs J Spiess AC 《Biotechnology and bioengineering》2008,101(1):27-38
For reactions using thiamine diphosphate (ThDP)-dependent enzymes many empirically-derived kinetic models exist. However, there is a lack of mechanistic kinetic models. This is especially true for the synthesis of symmetric 2-hydroxy ketones from two identical aldehydes, with one substrate acting as the donor and the other as the acceptor. In this contribution, a systematic approach for deriving such a kinetic model for thiamine diphosphate (ThDP)-dependent enzymes is presented. The derived mechanistic kinetic model takes this donor-acceptor principle into account by containing two K(m)-values even for identical substrate molecules. As example the stereoselective carbon-carbon coupling of two 3,5-dimethoxy-benzaldehyde molecules to (R)-3,3',5,5'-tetramethoxy-benzoin using benzaldehyde lyase (EC 4.1.2.38) from Pseudomonas fluorescens is studied. The model is derived using a model-based experimental analysis method which includes parameter estimation, model analysis, optimal experimental design, in silico experiments, sensitivity analysis and model revision. It is shown that this approach leads to a robust kinetic model with accurate parameter estimates and an excellent prediction capability. 相似文献
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Florine Dupeux Martin Rwer Gael Seroul Delphine Blot Jos A. Mrquez 《Acta Crystallographica. Section D, Structural Biology》2011,67(11):915-919
The identification of crystallization conditions for biological molecules largely relies on a trial‐and‐error process in which a number of parameters are explored in large screening experiments. Currently, construct design and sample formulation are recognized as critical variables in this process and often a number of protein variants are assayed for crystallization either sequentially or in parallel, which adds complexity to the screening process. Significant effort is dedicated to sample characterization and quality‐control experiments in order to identify at an early stage and prioritize those samples which would be more likely to crystallize. However, large‐scale studies relating crystallization success to sample properties are generally lacking. In this study, the thermal stability of 657 samples was estimated using a simplified Thermofluor assay. These samples were also subjected to automated vapour‐diffusion crystallization screening under a constant protocol. Analysis of the data shows that samples with an apparent melting temperature (Tm) of 318 K or higher crystallized in 49% of cases, while the crystallization success rate decreased rapidly for samples with lower Tm. Only 23% of samples with a Tm below 316 K produced crystals. Based on this analysis, a simple method for estimation of the crystallization likelihood of biological samples is proposed. This method is easy, rapid and consumes very small amounts of sample. The results of this assay can be used to determine optimal incubation temperatures for crystallization experiments or to prioritize certain constructs. More generally, this work provides an objective test that can contribute to making decisions in both focused and structural genomics crystallography projects. 相似文献
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Climate change has been shown to cause poleward range shifts of species. These shifts are typically demonstrated using presence–absence data, which can mask the potential changes in the abundance of species. Moreover, changes in the mean centre of weighted density of species are seldom examined, and comparisons between these two methods are even rarer. Here, we studied the change in the mean weighted latitude of density (MWLD) of 94 bird species in Finland, northern Europe, using data covering a north–south gradient of over 1000 km from the 1970s to the 2010s. The MWLD shifted northward on average 1.26 km yr?1, and this shift was significantly stronger in northern species compared to southern species. These shifts can be related to climate warming during the study period, because the annual temperature had increased more in northern Finland (by 1.7 °C) than in southern Finland (by 1.4 °C), although direct causal links cannot be shown. Density shifts of species distributed over the whole country did not differ from shifts in species situated on the edge of the species range in southern and northern species. This means that density shifts occur both in the core and on the edge of species distribution. The species‐specific comparison of MWLD values with corresponding changes in the mean weighted latitude using presence–absence atlas data (MWL) revealed that the MWLD moved more slowly than the MWL in the atlas data in the southern species examined, but more rapidly in the northern species. Our findings highlight that population densities are also moving rapidly towards the poles and the use of presence–absence data can mask the shift of population densities. We encourage use of abundance data in studies considering the effects of climate change on biodiversity. 相似文献
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Marcelo Sternberg Carly Golodets Mario Gutman Avi Perevolotsky Eugene D. Ungar Jaime Kigel Zalmen Henkin 《Global Change Biology》2015,21(5):1939-1950
A synthesis of a long‐term (19 years) study assessing the effects of cattle grazing on the structure and composition of a Mediterranean grassland in north‐eastern Israel is presented, with new insights into the response of the vegetation to grazing management and rainfall. We hypothesized that the plant community studied would be resistant to high grazing intensities and rainfall variability considering the combined long history of land‐use and unpredictable climatic conditions where this community evolved. Treatments included manipulations of stocking densities (moderate, heavy, and very heavy) and of grazing regimes (continuous vs. seasonal), in a factorial design. The effect of interannual rainfall variation on the expression of grazing impacts on the plant community was minor. The main effects of grazing on relative cover of plant functional groups were related to early vs. late seasonal grazing. Species diversity and equitability were remarkably stable across all grazing treatments. A reduction in tall grass cover at higher stocking densities was correlated with increased cover of less palatable groups such as annual and perennial thistles, as well as shorter and prostrate groups such as short annual grasses. This long‐term study shows that interannual fluctuations in plant functional group composition could be partly accounted for by grazing pressure and timing, but not by the measured rainfall variables. Grazing affected the dominance of tall annual grasses. However, the persistence of tall grasses and more palatable species over time, despite large differences in grazing pressure and timing, supports the idea that Mediterranean grasslands are highly resistant to prolonged grazing. Indeed, even under the most extreme grazing conditions applied, there were no signs of deterioration or collapse of the ecosystem. This high resistance to grazing intensity and interannual fluctuation in climatic conditions should favor the persistence of the plant community under forecasted increasing unpredictability due to climate change. 相似文献
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HARALD PAULI MICHAEL GOTTFRIED KARL REITER CHRISTIAN KLETTNER GEORG GRABHERR 《Global Change Biology》2007,13(1):147-156
High mountain ecosystems are defined by low temperatures and are therefore considered to react sensitively to climate warming. Responding to observed changes in plant species richness on high peaks of the European Alps, an extensive setup of 1 m × 1 m permanent plots was established at the alpine‐nival ecotone (between 2900 and 3450 m) on Mount Schrankogel, a GLORIA master site in the central Tyrolean Alps, Austria, in 1994. Recording was repeated in a representative selection of 362 quadrats in 2004. Ten years after the first recording, we observed an average change in vascular plant species richness from 11.4 to 12.7 species per plot, an increase of 11.8% (or of at least 10.6% at a 95% confidence level). The increase in species richness involved 23 species (about 43% of all taxa found at the ecotone), comprising both alpine and nival species and was pronouncedly higher in plots with subnival/nival vegetation than in plots with alpine grassland vegetation. Only three species showed a decrease in plot occupancy: one was an annual species, one was rare, and one a common nival plant that decreased in one part of the area but increased in the uppermost part. Species cover changed in relation to altitudinal preferences of species, showing significant declines of all subnival to nival plants, whereas alpine pioneer species increased in cover. Recent climate warming in the Alps, which has been twice as high as the global average, is considered to be the primary driver of the observed differential changes in species cover. Our results indicate an ongoing range contraction of subnival to nival species at their rear (i.e. lower) edge and a concurrent expansion of alpine pioneer species at their leading edge. Although this was expected from predictive distribution models and different temperature‐related habitat preferences of alpine and nival species, we provide first evidence on – most likely – warming‐induced species declines in the high European Alps. The projected acceleration of climate warming raises concerns that this phenomenon could become the major threat to biodiversity in high mountains. 相似文献
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Bessler C Schmitt J Maurer KH Schmid RD 《Protein science : a publication of the Protein Society》2003,12(10):2141-2149
alpha-Amylases, in particular, microbial alpha-amylases, are widely used in industrial processes such as starch liquefaction and pulp processes, and more recently in detergency. Due to the need for alpha-amylases with high specific activity and activity at alkaline pH, which are critical parameters, for example, for the use in detergents, we have enhanced the alpha-amylase from Bacillus amyloliquefaciens (BAA). The genes coding for the wild-type BAA and the mutants BAA S201N and BAA N297D were subjected to error-prone PCR and gene shuffling. For the screening of mutants we developed a novel, reliable assay suitable for high throughput screening based on the Phadebas assay. One mutant (BAA 42) has an optimal activity at pH 7, corresponding to a shift of one pH unit compared to the wild type. BAA 42 is active over a broader pH range than the wild type, resulting in a 5-fold higher activity at pH 10. In addition, the activity in periplasmic extracts and the specific activity increased 4- and 1.5-fold, respectively. Another mutant (BAA 29) possesses a wild-type-like pH profile but possesses a 40-fold higher activity in periplasmic extracts and a 9-fold higher specific activity. The comparison of the amino acid sequences of these two mutants with other homologous microbial alpha-amylases revealed the mutation of the highly conserved residues W194R, S197P, and A230V. In addition, three further mutations were found K406R, N414S, and E356D, the latter being present in other bacterial alpha-amylases. 相似文献
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