Simulated Microgravity Affects Growth of Escherichia coli and Recombinant β-d-Glucuronidase Production

Xiang, Lei; Qi, Feng; Dai, Dongfang; Chun, Li; Jiang, Yue

doi:10.1007/s12010-009-8836-0

Cited by 26 publications

(36 citation statements)

References 23 publications

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“…29 RPMs are commercially available by Mitsubishi Heavy Industries (Kobe, Japan) and Dutch Space (Leiden, The Netherlands), while various academic groups developed similar systems of 3D clinostats and RPMs. [35][36][37][38][39][40] Rotating wall vessel…”

Section: Random-positioning Machinementioning

confidence: 99%

Growing Tissues in Real and Simulated Microgravity: New Methods for Tissue Engineering

Grimm

Wehland

Pietsch

et al. 2014

Tissue Engineering Part B: Reviews

122

137

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Tissue engineering in simulated (s-) and real microgravity (r-mg) is currently a topic in Space medicine contributing to biomedical sciences and their applications on Earth. The principal aim of this review is to highlight the advances and accomplishments in the field of tissue engineering that could be achieved by culturing cells in Space or by devices created to simulate microgravity on Earth. Understanding the biology of three-dimensional (3D) multicellular structures is very important for a more complete appreciation of in vivo tissue function and advancing in vitro tissue engineering efforts. Various cells exposed to r-mg in Space or to s-mg created by a random positioning machine, a 2D-clinostat, or a rotating wall vessel bioreactor grew in the form of 3D tissues. Hence, these methods represent a new strategy for tissue engineering of a variety of tissues, such as regenerated cartilage, artificial vessel constructs, and other organ tissues as well as multicellular cancer spheroids. These aggregates are used to study molecular mechanisms involved in angiogenesis, cancer development, and biology and for pharmacological testing of, for example, chemotherapeutic drugs or inhibitors of neoangiogenesis. Moreover, they are useful for studying multicellular responses in toxicology and radiation biology, or for performing coculture experiments. The future will show whether these tissue-engineered constructs can be used for medical transplantations. Unveiling the mechanisms of microgravity-dependent molecular and cellular changes is an up-to-date requirement for improving Space medicine and developing new treatment strategies that can be translated to in vivo models while reducing the use of laboratory animals.

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Section: Random-positioning Machinementioning

confidence: 99%

Growing Tissues in Real and Simulated Microgravity: New Methods for Tissue Engineering

Grimm

Wehland

Pietsch

et al. 2014

Tissue Engineering Part B: Reviews

122

137

View full text Add to dashboard Cite

show abstract

“…Changes in cellular functions include cell clumping, shortened lag phase and prolonged exponential phase in the gene expression, and protein folding to alter the productivities of the secondary metabolites in bacteria (Escherichia coli, Pseudomonas aeruginosa) and yeast (S. cerevisiae) (Benoit et al 2006;Baker et al 2004;Crabbé et al 2011;Sheehan et al 2007). Only a few studies have shown that microgravity can impact the heterologous proteins production, such as enhanced expression of the recombinant β-glucuronidase in E. coli, and β-galactosidase and glycodelin in human cells (Xiang et al 2010;Stephen 2007). In contrast, some other studies indicated that SMG could not always promote protein expression (Boyle et al 1995).…”

Section: Discussionmentioning

confidence: 99%

Novel helper factors influencing recombinant protein production in Pichia pastoris based on proteomic analysis under simulated microgravity

Huangfu

Liu

et al. 2014

Appl Microbiol Biotechnol

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Microgravity and simulated microgravity (SMG) have quite significant effects on numerous microbial cellular processes. The effects of SMG on the production of recombinant proteins and transcription profiling in prokaryotic and eukaryotic expression host have been investigated. The present study showed that SMG significantly enhanced the specific productivities and activities of the reporter enzymes PGUS and AtXYN that were expressed in recombinant Pichia pastoris. Proteomic profiling revealed that 21 proteins were significantly up-regulated and 35 proteins were drastically down-regulated at the stationary phase, when the recombinant P. pastoris responded to SMG. Six strongly up-regulated genes, TPX, FBA, PGAM, ENO, SBA1, and AKR-E, involved in the oxidative stress response, methanol metabolism, glycolytic pathway, and protein folding, were selected to analyze their impacts on recombinant protein production by co-overexpression in the shaker flask fermentation. The co-overexpressed strains, particularly TPX, FBA, and PGAM, demonstrated promising results with approximately 2.46-fold, 1.58-fold, and 1.33-fold increases in the specific yields of PGUS compared to the control after 48 h of methanol induction, respectively. In the meantime, the corresponding PGUS specific activities were increased by 2.33-fold, 2.09-fold, and 1.32-fold, respectively. Thiol peroxidase (TPX), which is involved in the oxidative stress response, significantly influenced the transcriptional levels of the reporter gene PGUS. The present study provides valuable information for further exploration of the molecular mechanism of P. pastoris response to SMG and facilitates simulated microgravity for finding novel helper factors to rationally engineer the strains in normal fermentation by using proteomic studies.

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“…PGUS from Penicillium purpurogenum Li-3 screened in our previous work [17] can directly biosynthesize glycyrrhetinic acid monoglucuronide (GAMG) from glycyrrhizin (GL) (Fig. 2) [17,18]. GAMG is useful in clinical treatment of inflammatory diseases and is safer, more effective, and absorbable than GL [19].…”

Section: Rotationmentioning

confidence: 99%

Effects of Low-Shear Modeled Microgravity on the Characterization of Recombinant β-D-Glucuronidase Expressed in Pichia pastoris

Dai

et al. 2010

Appl Biochem Biotechnol

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In this study, we used a high-aspect-ratio vessel (HARV), which could model environment of microgravity on ground to investigate for the first time the effects of low-shear modeled microgravity (LSMMG) on the characterization of recombinant β-D-glucuronidase expressed in Pichia pastoris. The β-D-glucuronidase gene (GenBank accession no. EU095019) derived from Penicillium purpurogenum Li-3 encoding β-D-glucuronidase (PGUS) was expressed in P. pastoris GS115 in two different environments of LSMMG and normal gravity (NG). Results manifested that both LSMMG and NG conditions had insignificant effects on temperature and pH activity (optimal temperature and pH were 55 and 5.0 °C, respectively) and characteristic stability of recombinant PGUS. However, the catalytic activity of recombinant PGUS expressed under LSMMG was less affected by metal ions and EDTA as compared with that of NG. Furthermore, K (m) value of the recombinant PGUS expressed under LSMMG was nearly one fifth of that under NG (1.72 vs. 7.72), whereas catalytic efficiency (k (cat)/K (m)) of PGUS expressed under LSMMG (13.55) was 3.7 times higher than that of NG (3.61). The results initially reveal the significant alterations in catalytic properties of recombinant enzyme in response to LSMMG environment and have potential application in bioprocessing and biocatalysis.

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Simulated Microgravity Affects Growth of Escherichia coli and Recombinant β-d-Glucuronidase Production

Cited by 26 publications

References 23 publications

Growing Tissues in Real and Simulated Microgravity: New Methods for Tissue Engineering

Growing Tissues in Real and Simulated Microgravity: New Methods for Tissue Engineering

Novel helper factors influencing recombinant protein production in Pichia pastoris based on proteomic analysis under simulated microgravity

Effects of Low-Shear Modeled Microgravity on the Characterization of Recombinant β-D-Glucuronidase Expressed in Pichia pastoris

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