scholarly journals Isolated Microbial Single Cells and Resulting Micropopulations Grow Faster in Controlled Environments

2012 ◽  
Vol 78 (19) ◽  
pp. 7132-7136 ◽  
Author(s):  
Christian Dusny ◽  
Frederik Sven Ole Fritzsch ◽  
Oliver Frick ◽  
Andreas Schmid

ABSTRACTSingularized cells ofPichia pastoris,Hansenula polymorpha, andCorynebacterium glutamicumdisplayed specific growth rates under chemically and physically constant conditions that were consistently higher than those obtained in populations. This highlights the importance of single-cell analyses by uncoupling physiology and the extracellular environment, which is now possible using the Envirostat 2.0 concept.

2016 ◽  
Vol 82 (15) ◽  
pp. 4570-4583 ◽  
Author(s):  
Corinna Rebnegger ◽  
Tim Vos ◽  
Alexandra B. Graf ◽  
Minoska Valli ◽  
Jack T. Pronk ◽  
...  

ABSTRACTThe yeastPichia pastorisis a widely used host for recombinant protein production. Understanding its physiology at extremely low growth rates is a first step in the direction of decoupling product formation from cellular growth and therefore of biotechnological relevance. Retentostat cultivation is an excellent tool for studying microbes at extremely low specific growth rates but has so far not been implemented forP. pastoris. Retentostat feeding regimes were based on the maintenance energy requirement (mS) and maximum biomass yield on glucose (YX/Smax) estimated from steady-state glucose-limited chemostat cultures. Aerobic retentostat cultivation enabled reproducible, smooth transitions from a specific growth rate (μ) of 0.025 h−1to near-zero specific growth rates (μ < 0.001 h−1). At these near-zero specific growth rates, viability remained at least 97%. The value ofmSat near-zero growth rates was 3.1 ± 0.1 mg glucose per g biomass and h, which was 3-fold lower than themSestimated from faster-growing chemostat cultures. This difference indicated thatP. pastorisreduces its maintenance energy requirement at extremely low μ, a phenomenon not previously observed in eukaryotes. Intracellular levels of glycogen and trehalose increased, while μ progressively declined during retentostat cultivation. Transcriptional reprogramming toward zero growth included the upregulation of many transcription factors as well as stress-related genes and the downregulation of cell cycle genes. This study underlines the relevance of comparative analysis of maintenance energy metabolism, which has an important impact on large-scale industrial processes.IMPORTANCEThe yeastPichia pastorisnaturally lives on trees and can utilize different carbon sources, among them glucose, glycerol, and methanol. In biotechnology, it is widely used for the production of recombinant proteins. For both the understanding of life in its natural habitat and optimized production processes, a better understanding of cell physiology at an extremely low growth rate would be of extraordinary value. Therefore, we have grownP. pastorisin a retentostat, which allows the cultivation of metabolically active cells even at zero growth. Here we reached doubling times as long as 38 days and found thatP. pastorisdecreases its maintenance energy demand 3-fold during very slow growth, which enables it to survive with a much lower substrate supply than baker's yeast.


2012 ◽  
Vol 79 (5) ◽  
pp. 1646-1653 ◽  
Author(s):  
Dawn E. Holmes ◽  
Ludovic Giloteaux ◽  
Melissa Barlett ◽  
Milind A. Chavan ◽  
Jessica A. Smith ◽  
...  

ABSTRACTMolecular tools that can provide an estimate of thein situgrowth rate ofGeobacterspecies could improve understanding of dissimilatory metal reduction in a diversity of environments. Whole-genome microarray analyses of a subsurface isolate ofGeobacter uraniireducens, grown under a variety of conditions, identified a number of genes that are differentially expressed at different specific growth rates. Expression of two genes encoding ribosomal proteins,rpsCandrplL, was further evaluated with quantitative reverse transcription-PCR (qRT-PCR) in cells with doubling times ranging from 6.56 h to 89.28 h. Transcript abundance ofrpsCcorrelated best (r2= 0.90) with specific growth rates. Therefore, expression patterns ofrpsCwere used to estimate specific growth rates ofGeobacterspecies during anin situuranium bioremediation field experiment in which acetate was added to the groundwater to promote dissimilatory metal reduction. Initially, increased availability of acetate in the groundwater resulted in higher expression ofGeobacter rpsC, and the increase in the number ofGeobactercells estimated with fluorescentin situhybridization compared well with specific growth rates estimated from levels ofin situ rpsCexpression. However, in later phases, cell number increases were substantially lower than predicted fromrpsCtranscript abundance. This change coincided with a bloom of protozoa and increased attachment ofGeobacterspecies to solid phases. These results suggest that monitoringrpsCexpression may better reflect the actual rate thatGeobacterspecies are metabolizing and growing duringin situuranium bioremediation than changes in cell abundance.


2013 ◽  
Vol 79 (8) ◽  
pp. 2503-2511 ◽  
Author(s):  
Rudolf Schneebeli ◽  
Thomas Egli

ABSTRACTMembers of the genusListeriaare fastidious bacteria with respect to their nutritional requirements, and several minimal media described in the literature fail to support growth of allListeriaspp. Furthermore, strict limitation by a single nutrient, e.g., the carbon source, has not been demonstrated for any of the published minimal media. This is an important prerequisite for defined studies of growth and physiology, including “omics.” Based on a theoretical analysis of previously published mineral media forListeria, an improved, well-balanced growth medium was designed. It supports the growth, not only of all testedListeria monocytogenesstrains, but of all otherListeriaspecies, with the exception ofL. ivanovii. The growth performance ofL. monocytogenesstrain Scott A was tested in the newly designed medium; glucose served as the only carbon and energy source for growth, whereas neither the supplied amino acids nor the buffering and complexing components (MOPS [morpholinepropanesulfonic acid] and EDTA) supported growth. Omission of amino acids, trace elements, or vitamins, alone or in combination, resulted in considerably reduced biomass yields. Furthermore, we monitored the specific growth rates of variousListeriastrains cultivated in the designed mineral medium and compared them to growth in complex medium (brain heart infusion broth [BHI]). The novel mineral medium was optimized for the commonly used strainL. monocytogenesScott A to achieve optimum cell yields and maximum specific growth rates. This mineral medium is the first published synthetic medium forListeriathat has been shown to be strictly carbon (glucose) limited.


2001 ◽  
Vol 58 (2) ◽  
pp. 386-393 ◽  
Author(s):  
John A Sweka ◽  
Kyle J Hartman

Brook trout (Salvelinus fontinalis) were held in an artificial stream to observe the influence of turbidity on mean daily consumption and specific growth rates. Treatment turbidity levels ranged from clear (<3.0 nephelometric turbidity units (NTU)) to very turbid water (> 40 NTU). Observed mean daily specific consumption rates were standardized to the mean weight of all brook trout tested. Turbidity had no significant effect on mean daily consumption, but specific growth rates decreased significantly as turbidity increased. Brook trout in turbid water became more active and switched foraging strategies from drift feeding to active searching. This switch was energetically costly and resulted in lower specific growth rates in turbid water as compared with clear water. Bioenergetics simulations were run to compare observed growth with that predicted by the model. Observed growth values fell below those predicted by the model and the difference increased as turbidity increased. Abiotic factors, such as turbidity, which bring about changes in the activity rates of fish, can have implications for the accuracy of predicted growth by bioenergetics models.


Copeia ◽  
1992 ◽  
Vol 1992 (4) ◽  
pp. 1098 ◽  
Author(s):  
Alan B. Bolten ◽  
Karen A. Bjorndal ◽  
Janice S. Grumbles ◽  
David W. Owens

2014 ◽  
Vol 17 (2) ◽  
pp. 346-363 ◽  
Author(s):  
Wout Overkamp ◽  
Onur Ercan ◽  
Martijn Herber ◽  
Antonius J. A. van Maris ◽  
Michiel Kleerebezem ◽  
...  

2021 ◽  
Author(s):  
Sevtap Tırınk ◽  
Alper Nuhoğlu ◽  
Sinan Kul

Abstract This study encompasses investigation of treatment of pistachio processing industry wastewaters in a batch reactor under aerobic conditions, calculation of kinetic parameters and comparison of different inhibition models. The mixed microorganism culture used in the study was adapted to pistachio processing industry wastewaters for nearly one month and then concentrations from 50-1000 mg L− 1 of pistachio processing industry wastewaters were added to the medium and treatment was investigated in batch experiments. The Andrews, Han-Levenspiel, Luong and Aiba biokinetic equations were chosen for the correlations between the concentration of pistachio processing industry wastewaters and specific growth rates, and the kinetic parameters in these biokinetic equations were calculated. The µmax, Ks and Ki parameters, included in the Aiba biokinetic equation providing best fit among the other equations, had values calculated as 0.25 h− 1, 19 mg L− 1, and 516 mg L− 1, respectively.


2013 ◽  
Vol 10 (8) ◽  
pp. 5267-5280 ◽  
Author(s):  
F. H. Chang ◽  
E. C. Marquis ◽  
C. W. Chang ◽  
G. C. Gong ◽  
C. H. Hsieh

Abstract. Allometric scaling of body size versus growth rate and mortality has been suggested to be a universal macroecological pattern, as described by the metabolic theory of ecology (MTE). However, whether such scaling generally holds in natural assemblages remains debated. Here, we test the hypothesis that the size-specific growth rate and grazing mortality scale with the body size with an exponent of −1/4 after temperature correction, as MTE predicts. To do so, we couple a dilution experiment with the FlowCAM imaging system to obtain size-specific growth rates and grazing mortality of natural microphytoplankton assemblages in the East China Sea. This novel approach allows us to achieve highly resolved size-specific measurements that would be very difficult to obtain in traditional size-fractionated measurements using filters. Our results do not support the MTE prediction. On average, the size-specific growth rates and grazing mortality scale almost isometrically with body size (with scaling exponent ∼0.1). However, this finding contains high uncertainty, as the size-scaling exponent varies substantially among assemblages. The fact that size-scaling exponent varies among assemblages prompts us to further investigate how the variation of size-specific growth rate and grazing mortality can interact to determine the microphytoplankton size structure, described by normalized biomass size spectrum (NBSS), among assemblages. We test whether the variation of microphytoplankton NBSS slopes is determined by (1) differential grazing mortality of small versus large individuals, (2) differential growth rate of small versus large individuals, or (3) combinations of these scenarios. Our results indicate that the ratio of the grazing mortality of the large size category to that of the small size category best explains the variation of NBSS slopes across environments, suggesting that higher grazing mortality of large microphytoplankton may release the small phytoplankton from grazing, which in turn leads to a steeper NBSS slope. This study contributes to understanding the relative importance of bottom-up versus top-down control in shaping microphytoplankton size structure.


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