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1.
McCabe BK Kuek C Gordon GL Phillips MW 《Journal of industrial microbiology & biotechnology》2003,30(4):205-209
Two anaerobic fungi, one a monocentric strain (Piromyces sp. KSX1) and the other a polycentric strain (Orpinomyces sp. 478P1), were immobilised in calcium alginate beads and cultured in sequential batches where spent medium (containing
0.25% cellobiose) was repeatedly drained and replaced. β-Glucosidase production with KSX1 was maintained for 45 days over
six repeated batch cultures yielding a maximum level of 107 mIU/ml. For 478P1, β-glucosidase production was maintained for
30 days over four repeated batches yielding a maximum level of 34 mIU/ml. Although repeat-batch cultures of KSX1 produced
more β-glucosidase than strain 478P1, the maximum specific β-glucosidase produced from these immobilised cultures was similar.
The immobilised polycentric strain proved to be operationally superior to strain KSX1, as strain 478P1 did not produce any
growth in the culture liquor.
Electronic Publication 相似文献
2.
Carl Brunius Lin Shi Rikard Landberg 《Metabolomics : Official journal of the Metabolomic Society》2016,12(11):173
Introduction
Liquid chromatography-mass spectrometry (LC-MS) is a commonly used technique in untargeted metabolomics owing to broad coverage of metabolites, high sensitivity and simple sample preparation. However, data generated from multiple batches are affected by measurement errors inherent to alterations in signal intensity, drift in mass accuracy and retention times between samples both within and between batches. These measurement errors reduce repeatability and reproducibility and may thus decrease the power to detect biological responses and obscure interpretation.Objective
Our aim was to develop procedures to address and correct for within- and between-batch variability in processing multiple-batch untargeted LC-MS metabolomics data to increase their quality.Methods
Algorithms were developed for: (i) alignment and merging of features that are systematically misaligned between batches, through aggregating feature presence/missingness on batch level and combining similar features orthogonally present between batches; and (ii) within-batch drift correction using a cluster-based approach that allows multiple drift patterns within batch. Furthermore, a heuristic criterion was developed for the feature-wise choice of reference-based or population-based between-batch normalisation.Results
In authentic data, between-batch alignment resulted in picking 15 % more features and deconvoluting 15 % of features previously erroneously aligned. Within-batch correction provided a decrease in median quality control feature coefficient of variation from 20.5 to 15.1 %. Algorithms are open source and available as an R package (‘batchCorr’).Conclusions
The developed procedures provide unbiased measures of improved data quality, with implications for improved data analysis. Although developed for LC-MS based metabolomics, these methods are generic and can be applied to other data suffering from similar limitations.3.
Proteomics analysis of a long‐term survival strain of Escherichia coli K‐12 exhibiting a growth advantage in stationary‐phase (GASP) phenotype
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Assunta Gagliardi Egidio Lamboglia Laura Bianchi Claudia Landi Alessandro Armini Silvia Ciolfi Luca Bini Laura Marri 《Proteomics》2016,16(6):963-972
The aim of this work was the functional and proteomic analysis of a mutant, W3110 Bgl+/10, isolated from a batch culture of an Escherichia coli K‐12 strain maintained at room temperature without addition of nutrients for 10 years. When the mutant was evaluated in competition experiments in co‐culture with the wild‐type, it exhibited the growth advantage in stationary phase (GASP) phenotype. Proteomes of the GASP mutant and its parental strain were compared by using a 2DE coupled with MS approach. Several differentially expressed proteins were detected and many of them were successful identified by mass spectrometry. Identified expression‐changing proteins were grouped into three functional categories: metabolism, protein synthesis, chaperone and stress responsive proteins. Among them, the prevalence was ascribable to the “metabolism” group (72%) for the GASP mutant, and to “chaperones and stress responsive proteins” group for the parental strain (48%). 相似文献
4.
Beauvericin (BEA) is a cyclic hexadepsipeptide mycotoxin with notable phytotoxic and insecticidal activities. Fusarium redolens Dzf2 is a highly BEA-producing fungus isolated from a medicinal plant. The aim of the current study was to develop a simple
and valid kinetic model for F. redolens Dzf2 mycelial growth and the optimal fed-batch operation for efficient BEA production. A modified Monod model with substrate
(glucose) and product (BEA) inhibition was constructed based on the culture characteristics of F. redolens Dzf2 mycelia in a liquid medium. Model parameters were derived by simulation of the experimental data from batch culture.
The model fitted closely with the experimental data over 20–50 g l−1 glucose concentration range in batch fermentation. The kinetic model together with the stoichiometric relationships for biomass,
substrate and product was applied to predict the optimal feeding scheme for fed-batch fermentation, leading to 54% higher
BEA yield (299 mg l−1) than in the batch culture (194 mg l−1). The modified Monod model incorporating substrate and product inhibition was proven adequate for describing the growth kinetics
of F. redolens Dzf2 mycelial culture at suitable but not excessive initial glucose levels in batch and fed-batch cultures. 相似文献
5.
A flexible process monitoring method was applied to industrial pilot plant cell culture data for the purpose of fault detection and diagnosis. Data from 23 batches, 20 normal operating conditions (NOC) and three abnormal, were available. A principal component analysis (PCA) model was constructed from 19 NOC batches, and the remaining NOC batch was used for model validation. Subsequently, the model was used to successfully detect (both offline and online) abnormal process conditions and to diagnose the root causes. This research demonstrates that data from a relatively small number of batches (approximately 20) can still be used to monitor for a wide range of process faults. 相似文献
6.
Fungal morphology and fragmentation behavior in a fed-batch Aspergillus oryzae fermentation at the production scale 总被引:1,自引:0,他引:1
Li ZJ Shukla V Fordyce AP Pedersen AG Wenger KS Marten MR 《Biotechnology and bioengineering》2000,70(3):300-312
It is well known that high-viscosity fermentation broth can lead to mixing and oxygen mass transfer limitations. The seemingly obvious solution for this problem is to increase agitation intensity. In some processes, this has been shown to damage mycelia, affect morphology, and decrease product expression. However, in other processes increased agitation shows no effect on productivity. While a number of studies discuss morphology and fragmentation at the laboratory and pilot scale, there are relatively few publications available for production-scale fungal fermentations. The goal of this study was to assess morphology and fragmentation behavior in large-scale, fed-batch, fungal fermentations used for the production of protein. To accomplish this, a recombinant strain of Aspergillus oryzae was grown in 80 m(3) fermentors at two different gassed, impeller power-levels (one 50% greater than the other). Impeller power is reported as energy dissipation/circulation function (EDCF) and was found to have average values of 29.3 +/- 1.0 and 22.0 +/- 0.3 kW m(-3) s(-1) at high and low power levels, respectively. In all batches, biomass concentration profiles were similar and specific growth rate was < 0.03 h(-1). Morphological data show hyphal fragmentation occurred by both shaving-off of external clump hyphae and breakage of free hyphae. The fragmentation rate constant (k(frag)), determined using a first-order model, was 5.90 and 5.80 h(-1) for high and low power batches, respectively. At the end of each batch, clumps accounted for only 25% of fungal biomass, most of which existed as small, sparsely branched, free hyphal elements. In all batches, fragmentation was found to dominate fungal growth and branching. We speculate that this behavior was due to slow growth of the culture during this fed-batch process. 相似文献
7.
Conversion of glucose‐xylose mixtures to pyruvate using a consortium of metabolically engineered Escherichia coli
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Two strains of Escherichia coli were engineered to accumulate pyruvic acid from two sugars found in lignocellulosic hydrolysates by knockouts in the aceE, ppsA, poxB, and ldhA genes. Additionally, since glucose and xylose are typically consumed sequentially due to carbon catabolite repression in E. coli, one strain (MEC590) was engineered to grow only on glucose while a second strain (MEC589) grew only on xylose. On a single substrate, each strain generated pyruvate at a yield of about 0.60 g/g in both continuous culture and batch culture. In a glucose‐xylose mixture under continuous culture, a consortium of both strains maintained a pyruvate yield greater than 0.60 g/g when three different concentrations of glucose and xylose were sequentially fed into the system. In a fed‐batch process, both sugars in a glucose‐xylose mixture were consumed simultaneously to accumulate 39 g/L pyruvate in less than 24 h at a yield of 0.59 g/g. 相似文献
8.
Nowadays, bioprocesses are developed or optimized on small scale. Also, vinegar industry is motivated to reinvestigate the established repeated batch fermentation process. As yet, there is no small‐scale culture system for optimizing fermentation conditions for repeated batch bioprocesses. Thus, the aim of this study is to propose a new shaken culture system for parallel repeated batch vinegar fermentation. A new operation mode — the flushing repeated batch — was developed. Parallel repeated batch vinegar production could be established in shaken overflow vessels in a completely automated operation with only one pump per vessel. This flushing repeated batch was first theoretically investigated and then empirically tested. The ethanol concentration was online monitored during repeated batch fermentation by semiconductor gas sensors. It was shown that the switch from one ethanol substrate quality to different ethanol substrate qualities resulted in prolonged lag phases and durations of the first batches. In the subsequent batches the length of the fermentations decreased considerably. This decrease in the respective lag phases indicates an adaptation of the acetic acid bacteria mixed culture to the specific ethanol substrate quality. Consequently, flushing repeated batch fermentations on small scale are valuable for screening fermentation conditions and, thereby, improving industrial‐scale bioprocesses such as vinegar production in terms of process robustness, stability, and productivity. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1158–1168, 2013 相似文献
9.
Fujino Y Kikuchi K Nakamura Y Kobayashi H Yonemura I Suzuki M Misumi K Nagai T 《Theriogenology》2007,67(2):413-422
The viability or developmental ability of porcine embryos after slow-freezing and thawing differs depending on the embryonic stage or the batch, which is defined as a group of embryos obtained from one donor at one time. We froze porcine blastocysts in batches and assessed their cryotolerance by using two expanded blastocysts (EBs) as samples to predict the developmental potential of other blastocysts from the same batch at different stages. Two EBs from the same batch that had been separately frozen were thawed and cultured in vitro for 48 h to examine their in vitro ability to develop to the hatched blastocyst stage. Thereafter, each batch was assigned to Grade A, B, or C according to the viability of the two EBs, i.e., 100% viability (2/2: number of hatched blastocysts/number of cultured EBs) was Grade A; 50% (1/2) was Grade B; and 0% (0/2) was Grade C. The viability of EBs after freeze-thawing and in vitro culture varied depending on the batch and was lower (31.0+/-10.2%, mean+/-S.E.M.; P<0.01) than that of unfrozen controls (96.8+/-2.3%). The viability of frozen-thawed hatched blastocysts (HBs) did not differ among the graded batches, but the blastocyst diameter decreased (from 409 to 326 microm) as the batch grade decreased (from A to C). When both EBs and HBs from batches of the same grade were transferred to recipients (average 11.7 EBs and 16.0 HBs per recipient), the rate of pregnancy and farrowing in recipients decreased (from 77.8% to 0%) and the number of piglets obtained decreased (from 15.3 to 0) as the batch grade decreased. However, when not only frozen-thawed EBs from Grade B or C batches, but also four helper embryos at the morula to early blastocyst stage (which were expected to support the pregnancy) were transferred, the number of piglets generated was higher from EBs from Grade B batches (16.0) than from EBs from Grade C batches (0.0). When frozen-thawed HBs and helper embryos were transferred, the number of piglets generated was higher from HBs from Grade B batches (12.7) than that from HBs from Grade C batches (1.9). After slow-freezing of porcine blastocysts, their rate of survival to the piglet stage differs batchwise, and in vitro viability assessment of sample EBs after freezing and thawing may help in assessing the post-freezing and post-thawing developmental potential of other blastocysts at different stages from the same batch. 相似文献
10.
Summary The isolation of silver-resistant, silver-accumulating bacteria is reported. Following the screening of a number of environmental sources, silver-resistant Enterobacteriaceae were isolated from both sewage and photographic processing effluent. The level of resistance to silver and other heavy metals was determined for a selection of these isolates and, together with preliminary accumulation data derived from batch culture studies, one isolate, a strain of Citrobacter intermedius, was selected for further examination. The effect of silver concentration on batch culture growth of this organism was also investigated. 相似文献
11.
Repeated-batch production of kojic acid in a cell-retention fermenter using Aspergillus oryzae M3B9 总被引:1,自引:0,他引:1
Wan HM Chen CC Giridhar R Chang TS Wu WT 《Journal of industrial microbiology & biotechnology》2005,32(6):227-233
A cell-retention fermenter was used for the pilot-scale production of kojic acid using an improved strain of Aspergillus oryzae in repeated-batch fermentations. Among the various carbon and nitrogen sources used, sucrose and yeast extract promoted pellet morphology of fungi and higher kojic acid production. Repeated-batch culture using a medium replacement ratio of 75% gave a productivity of 5.3 g L–1 day–1 after 11.5 days of cultivation. While batch culture in shake-flasks resulted in a productivity of 5.1 g L–1 day–1, a productivity of 5 g L–1 day–1 was obtained in a pilot-scale fermenter. By converting the batch culture into repeated batches, the non-productive downtime of cleaning, filling and sterilizing the fermenter between each batch were eliminated, thereby increasing the kojic acid productivity. 相似文献
12.
Reference strains (2) and 29 isolates ofAeromonas spp. from clinical material and environmental specimens were characterised in traditional biochemical tests, and in pyrolysis mass spectrometry, which gives data reflecting whole-cell composition. Numerical taxonomic analyses of the data sets were compared with conventional identification at species level, and pathogenic potential, as inferred from the origin of the isolates. Clustering with conventional test reaction patterns showed, for each of the species represented, a clearly defined core group of typical isolates, surrounded by a halo of aberrant strains. One further cluster comprised strains intermediate betweenA. caviae andA. hydrophila, and one strain was grossly atypical in both analyses. Clustering from pyrolysis data corresponded less well with species identification. Broadly, the biochemical division between core and halo strains was supported in pyrolysis forA. caviae andA. sobria, but the main group ofA. hydrophila in pyrolysis comprised strains clustering in the core and halo groups of this species, and three strains intermediate betweenA. hydrophila andA. caviae in biochemical tests. Two further pyrolysis clusters comprised core and halo strains ofA. hydrophila. However, pyrolysis clustering correlated well with inferred pathogenicity, showing four clusters of probable pathogens, six clusters of probable nonpathogens, and one two member cluster of doubtful status. Most strains that clustered in the species haloes, or in species-intermediate groups in biochemical tests, were non-human isolates, or were isolated in the absence of symptomatic infection. The correlation of inferred pathogenicity with biochemical clustering was poorer than that with pyrolysis clustering.Abbreviations CTRP
conventional test reaction pattern
- PyMS
pyrolysis mass spectrometry 相似文献
13.
Summary The recycled solid-state surface fermentation (SSF) culture ofAspergillus niger KCU520 was used for repeated batch production of citric acid from sugarcane molasses. The rate of citric acid production was doubled, reducing the fermentation time to half, compared to the normal single cycle batch submerged or surface fermentation process. About 80% sugar was converted to citric acid in five-day batch fermentation and three batches were carried out with the same fungal mat without any significant loss of productivity. 相似文献
14.
This study addressed the problem of maximizing cell mass and monoclonal antibody production from a fed-batch hybridoma cell culture. We hypothesized that inaccuracies in the process model limited the mathematical optimization. On the basis of shaker flask data, we established a simple phenomenological model with cell mass and lactate production as the controlled variables. We then formulated an optimal control algorithm, which calculated the process-model mismatch at each sampling time, updated the model parameters, and re-optimized the substrate concentrations dynamically throughout the time course of the batch. Manipulated variables were feed rates of glucose and glutamine. Dynamic parameter adjustment was done using a fuzzy logic technique, while a heuristic random optimizer (HRO) optimized the feed rates. The parameters selected for updating were specific growth rate and the yield coefficient of lactate from glucose. These were chosen by a sensitivity analysis. The cell mass produced using dynamic optimization was compared to the cell mass produced for an unoptimized case, and for a one-time optimization at the beginning of the batch. Substantial improvements in reactor productivity resulted from dynamic re-optimization and parameter adjustment. We demonstrated first that a single offline optimization of substrate concentration at the start of the batch significantly increased the yield of cell mass by 27% over an unoptimized fermentation. Periodic optimization online increased yield of cell mass per batch by 44% over the single offline optimization. Concomitantly, the yield of monoclonal antibody increased by 31% over the off-line optimization case. For batch and fed-batch processes, this appears to be a suitable arrangement to account for inaccuracies in process models. This suggests that implementation of advanced yet inexpensive techniques can improve performance of fed-batch reactors employed in hybridoma cell culture. 相似文献
15.
Park WS Park SJ Han SJ Lee J Kim DS Kim JH Kim BW Lee J Sim SJ 《Journal of microbiology and biotechnology》2007,17(7):1208-1212
Production of extracellular epothilone B, one of the potent anticancer agents, by free and immobilized Sorangium cellulosum was studied using the repeated batch culture process. The concentration of alginate used in immobilization was directly related to the mass transfer rate of nutrients, mechanical stability, and the epothilone B production yield. With the optimized 3% (w/v) calcium alginate carrier, a prolonged repeated batch culture was investigated for the 5 repeated batches for 24 days. The maximum productivity of epothilone B obtained from the alginate-immobilized cells was 5.03 mg/l/day, which is 3 times higher than that of free cells (1.68 mg/l/day). 相似文献
16.
Nallet S Fornelli L Schmitt S Parra J Baldi L Tsybin YO Wurm FM 《New biotechnology》2012,29(4):471-476
In this study, a recombinant monoclonal IgG antibody was produced by transient gene expression (TGE) in suspension-adapted HEK-293E cells. The objective of the study was to determine the variation in recombinant IgG yield and glycosylation in ten independent transfections. In a ten-day batch process, the variation in transient IgG yield in the ten batches was less than 30% with the specific productivity averaging 20.2 ± 2.6 pg/cell/day. We characterized the N-glycosylation profile of each batch of affinity-purified IgG by intact protein and bottom-up mass spectrometry. Four major glycans were identified at Asn(297) in the ten batches with the maximum relative deviation for a single glycoform being 2.5%. In addition, within any single transfection there was little variation in glycoforms over the ten-day culture. Our experimental data indicate that with TGE, the production of recombinant IgG with little batch-to-batch variation in volumetric yield and protein glycosylation is feasible, even in a non-instrumented cultivation system as described here. 相似文献
17.
BIO - 《Biologicals》2002,30(4):277
In Europe, the target animal safety test (TAST) is stipulated by 52 European Pharmacopoeia monographs, by three European Union (EU) Directives and a number of EU guidelines as a routine test for veterinary immunologicals, to be carried out on the finished product. TAST data from seven European Official Member States Control Laboratories (OMCLs) and 14 manufacturers were retrospectively analysed. During 1994–1997, 11 185 vaccine batches had been submitted for batch release, and the OMCLs had tested 670 batches in the TAST (665 passed, 4 passed after retesting, 1 failed). In total, 82 of these batches were not released; however, in only one case this was due to failure in the TAST. The data received from the 14 manufacturers covered the years from 1997 to 1999. 11 386 batches were tested in the TAST, of which 215 passed after retesting and 7 failed. Although only 30% of the OMCLs provided data and the data of the manufacturers are not complete they clearly indicate that the TAST does not contribute to the safety of veterinary vaccines and should therefore not be required as a routine batch test. In cases, where it appears to be necessary, detailed guidance on the test design and evaluation should be given. Copyright 2002 Published by Elsevier Science Ltd on behalf of The International Association for Biologicals. 相似文献
18.
A miniaturized reactor system with on‐line measurement of respiration rates by membrane inlet mass spectrometry was applied for the on‐line metabolic flux analysis at different phases of a 1.2 L batch culture of lysine‐producing Corynebacterium glutamicum. For this purpose, cells taken from the batch culture were transferred into the 12 mL mini reactor, and incubated for 15 min with [1‐18O]glucose. Quantification of oxygen uptake rate and CO2 mass isotopomer production rates in combination with a simple metabolic model allowed the estimation of the flux partitioning ratio between the pentose phosphate pathway and glycolysis during the process. The relative flux into the pentose pathway increased during growth, and reached maxima at 11 and 17 h cultivation time coinciding with maxima of the differential lysine yield. The developed system is a promising tool for determination of metabolic flux dynamics in industrially relevant batch and fed‐batch cultures. 相似文献
19.
The total protein content and cell size distribution of recombinantSaccharomyces cerevisiae cells were analyzed by flow cytometry. The recombinant strain containing a regulatableSUC2 promoter and the host strain were compared when grown under similar conditions in a batch culture. Recombinant and host cells
maintained similar size and total protein content while cloned-gene expression was repressed by glucose levels greater than
0.2% (w/v). Following derepression, recombinant cells demonstrated a mean total protein content and mean cell size 1.5–2 times
greater than that of the host cells. In addition, these simple flow cytometric measurements of the changes in cell size and
total protein content were found to closely follow diauxic growth ofSaccharomyces cerevisiae in batch culture. 相似文献
20.
P. Fouchet C. Manin H. Richard G. Frelat J. N. Barbotin 《Applied microbiology and biotechnology》1994,41(5):584-590
Flow cytometry has been used to study the contents of macromolecular compounds and light-scatter parameters in batch and continuous cultures of a recombinant Escherichia coli strain that forms protein inclusion bodies. Changes in relative DNA and RNA contents and cell mass as estimated by forward-angle light scatter were detected and tightly correlated in batch culture. In addition, heterogeneity of wide-angle light scatter (WALS), which we related to the presence of cellular inclusion bodies, was observed. In contrast, the relative RNA content and cell mass did not change during continuous culture, and homogeneity of WALS was found. In addition, unexpected changes in relative DNA content were observed after 67 h of culture, indicating a change in bacterial physiology. 相似文献