Quantitative characterization of the regulation of the synthesis of alcohol oxidase and of the expression of recombinant avidin in a Pichia pastoris Mut+ strain

Jungo, Carmen; Rérat, Claude; Marison, I. W.; Stockar, Urs von

doi:10.1016/j.enzmictec.2006.01.027

Cited by 59 publications

(51 citation statements)

References 36 publications

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“…increasingly or decreasingly linear, hyperbolic or bell-shaped [e.g. (Bhatacharya et al, 2007, Curvers et al, 2001b, Graslund et al, 2008, Hang et al, 2008, Jungo et al, 2006, Kobayashi et al, 2000, Min et al, 2010, Potgieter et al, 2010, Schenk et al, 2008, Zhang et al, 2005]. Nevertheless, for strains containing the GAP-promoter, product formation was found to increase with specific growth rates almost up to µ max (Khasa et al, 2007, Maurer et al, 2006.…”

Section: Production Kineticsmentioning

confidence: 94%

“…Typically, production kinetics controlled by the AOX1-promotor, utilising methanol or mixtures of substrates with methanol, were investigated for µ < 0.08 h -1 (Curvers et al, 2001b, Zhang et al, 2005 or even µ < 0.03 h -1 (Hang et al, 2008, Kobayashi et al, 2000, Min et al, 2010, Potgieter et al, 2010. Only two kinetic-relationships with low q p -values (< 0.03 mg l -1 h -1 ) were investigated up to a µ of 0.14 h -1 (Jungo et al, 2006, Schenk et al, 2008. For processes in which production is controlled by the GAP-promoter, utilising glucose as a growth and energy substrate, specific growth rates of up to 0.2 h -1 were investigated (Khasa et al, 2007).…”

Section: Production Kineticsmentioning

confidence: 99%

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Cultivation strategies to enhance productivity of Pichia pastoris: A review

Looser

Brühlmann

Bumbak

et al. 2015

Biotechnology Advances

297

268

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Pichia pastoris, a methylotrophic yeast, is an established system for the production of heterologous proteins, particularly biopharmaceuticals and industrial enzymes. To maximise and optimise the production of recombinant products, recent molecular research has focused on numerous issues including the design of expression vectors, optimisation of gene copy number, co-expression of secretory proteins such as chaperones, engineering of glycosylation and secretory pathways, etc. However, the physiological effects of different cultivation strategies are often difficult to separate from the molecular effects of the gene construct (e.g., cellular stress through over-expression or incorrect post-translational processing). Hence, overall system optimisation is difficult, even though it is urgently required in order to describe and understand the behaviour of new molecular constructs. This review focuses on particular aspects of recombinant protein production related to variations in biomass growth and their implications for strain design and screening, as well as on the concept of rational comparisons between cultivation systems for the development of specific production processes in bioreactors. The relationship between specific formation rates of secreted recombinant proteins, qp, and specific growth rates, μ, has been analysed in a conceptual attempt to compare different systems, particularly those based on AOX1/methanol and GAP/glucose, and this has now evolved into a pivotal concept for bioprocess engineering of P. pastoris.

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Section: Production Kineticsmentioning

confidence: 94%

Section: Production Kineticsmentioning

confidence: 99%

Cultivation strategies to enhance productivity of Pichia pastoris: A review

Looser

Brühlmann

Bumbak

et al. 2015

Biotechnology Advances

297

268

View full text Add to dashboard Cite

show abstract

“…Steady or increasing biomass yields coupled with higher metabolic heat a Q T , total heat released by the microbial growth reaction as measured by the VP-ITC instrument (n ϭ 3); P max , maximum heat flow rate of the microbial growth reaction (n ϭ 3); t max , time to reach peak of power/time curve (n ϭ 3); k, metabolic rate constant (n ϭ 3); yield, carbon moles biomass divided by the carbon moles glucose consumed (n ϭ 3); Q T /C-mol biomass, total heat released per carbon mole biomass (cal/C-mol ϫ 10 output were also observed previously by von Stockar and Liu (47) when they analyzed the thermokinetic profiles of microbes growing on increasingly electron-poor substrates. The enthalpy of combustion and the elemental biomass composition changed when cells were grown on these substrates (9,22,47). Perhaps, the wild type and KT1 manufacture a proportionally larger amount of polysaccharides than protein under conditions of metal stress (especially KT1, which lacks CzcCBA1 expression), lowering the total amount of stored energy per carbon mole of biomass and freeing up more energy to be lost as heat (45).…”

Section: Discussionmentioning

confidence: 99%

Use of Microcalorimetry To Determine the Costs and Benefits to Pseudomonas putida Strain KT2440 of Harboring Cadmium Efflux Genes

Gibbons

Feris

McGuirl

et al. 2011

Appl Environ Microbiol

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A novel microcalorimetric approach was used to analyze the responses of a metal-tolerant soil bacterium (Pseudomonas putida strain KT2440) to metal resistance gene deletions in cadmium-amended media. As hypothesized, under cadmium stress, the wild-type strain benefited from the resistance genes by entering the exponential growth phase earlier than two knockout strains. In the absence of cadmium, strain KT1, carrying a deletion in the main component (czcA1) of a Cd/Zn chemiosmotic efflux transporter (CzcCBA1), grew more efficiently than the wild type and released ϳ700 kJ (per mole of biomass carbon) less heat than the wild-type strain, showing the energetic cost of maintaining CzcCBA1 in the absence of cadmium. A second mutant strain (KT4) carrying a different gene deletion, ⌬cadA2, which encodes the main Cd/Pb efflux transporter (a P-type ATPase), did not survive beyond moderate cadmium concentrations and exhibited a decreased growth yield in the absence of cadmium. Therefore, CadA2 plays an essential role in cadmium resistance and perhaps serves an additional function. The results of this study provide direct evidence that heavy metal cation efflux mechanisms facilitate shorter lag phases in the presence of metals and that the maintenance and expression of tolerance genes carry quantifiable energetic costs and benefits.Human activities have contaminated ecosystems around the world with persistent heavy metals (30). Metal-associated stress alters microbial community structure and metabolism (14,15,40) and therefore affects higher trophic interactions. Metal efflux pumps exploit the cellular pool of energy (ATP and chemiosmotic gradients) to pump metal ions out of a cell. Consequently, they should encumber the host organism with the energetic cost of maintaining and expressing the genes coding for them (10,32,42). The overall cost of encoding these traits, while necessary for survival in a contaminated environment, should decrease fitness in environments with low concentrations of metals (43). The energetic costs of maintaining metal resistance genes in the absence of bioavailable heavy metals in bacteria are generally assumed but apparently have never been directly quantified.The main toxic effect of cadmium on bacterial cells is to bind to and denature protein (33, 34). Cadmium-protein binding induces a cascade of other putative changes in the cell, such as the release of redox-active metals bound to proteins and the general disruption of many protein-mediated processes (20,44). Efflux pumps are a way of directly and efficiently dealing with cadmium, by transporting it out of the cell. Other cellular responses to cadmium involve the large-scale synthesis of molecules, such as glutathione, polysaccharides, and chaperone proteins, to bind cadmium and cope with cadmium-induced protein denaturation (13,16,20,33).Most bacteria nonspecifically import cadmium into the cell along with other divalent metal ions (31). The result can be seen in the evolution of multiple cadmium resistance mechanisms in bacteria (44). Mode...

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“…Recombinant protein production is growth associated (Jungo et al 2006;Hohenblum et al 2003;Zhang et al 2000). This can be seen in IDShr and LRO production (Figs.…”

Section: Resultsmentioning

confidence: 90%

“…The production of recombinant protein I is considered associated to the viable biomass growth (Jungo et al 2006;Cunha et al 2004;Hohenblum et al 2003;Zhang et al 2000).…”

Section: Viable Biomassmentioning

confidence: 99%

A simple structured model for recombinant IDShr protein production in Pichia pastoris

et al. 2008

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A simple structured model is proposed for the methanol production phase of the iduronate 2-sulphate sulfatase recombinant enzyme (IDShr) in Pichia patoris Mut(+). The model is mainly focused in oxidative stress phenomenon due to methanol consumption and based on extracellular experimental information and the basic knowledge of methanol metabolism in Pichia pastoris yeast (P. pastoris). The model's prediction shows a reasonable accuracy as compared with the experimental data. Likewise, it was proved that this model is able to simulate the production of other recombinant protein in P. pastoris.

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Quantitative characterization of the regulation of the synthesis of alcohol oxidase and of the expression of recombinant avidin in a Pichia pastoris Mut+ strain

Cited by 59 publications

References 36 publications

Cultivation strategies to enhance productivity of Pichia pastoris: A review

Cultivation strategies to enhance productivity of Pichia pastoris: A review

Use of Microcalorimetry To Determine the Costs and Benefits to Pseudomonas putida Strain KT2440 of Harboring Cadmium Efflux Genes

A simple structured model for recombinant IDShr protein production in Pichia pastoris

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