Low cost production of perdeuterated biomass using methylotrophic yeasts

Haon, Stéphane; Augé, Sophie; Tropis, Marielle; Milon, Alain; Lindley, Nic D.

doi:10.1002/jlcr.2580331108

Cited by 16 publications

(9 citation statements)

References 25 publications

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“…Expression of 8-and ~-opioid receptors which are highly homologous to ~t-opioid receptors will certainly follow. These results pave the way for large-scale production of/a-opioid receptors, stable isotope labelling [7] for NMR or neutron diffraction experiments, and for the determination of the conformations of peptidic ligands bound to the receptor and eventually of the receptor itself.…”

Section: Resultsmentioning

confidence: 89%

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Expression and pharmacological characterization of the human μ‐opioid receptor in the methylotrophic yeast Pichia pastoris

et al. 1996

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Section: Resultsmentioning

confidence: 89%

“…Produced proteins are usually properly folded because P. pastoris provides an intracellular folding machinery similar to that of mammalian cells. Finally, P. pastoris can be grown in minimal medium to obtain 2H-, 15N-and laC-labelled proteins [7].…”

Section: Introductionmentioning

confidence: 99%

Expression and pharmacological characterization of the human μ‐opioid receptor in the methylotrophic yeast Pichia pastoris

et al. 1996

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“…D 2 O is much less toxic to microorganisms than to multicellular creatures. After a period of adaptation, a number of bacteria and algae can grow in pure D 2 O, although usually more slowly than in H 2 O (Thomson 1963;Katz 1960Katz , 1965Crespi 1982;Unno et al 1989;Vanatulu et al 1993;Haon et al 1993;Kushner et al 1997). More complex cells, such as protozoa, can grow in 70% or higher D 2 O (Thomson 1963).…”

Section: Effects Of D 2 O On Proteins Cells and Tissuesmentioning

confidence: 99%

Pharmacological uses and perspectives of heavy water and deuterated compounds

Kushner¹,

Baker²,

Dunstall³

1999

Can. J. Physiol. Pharmacol.

253

177

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Since the discovery of D20 (heavy water) and its use as a moderator in nuclear reactors, its biological effects have been extensively, although seldom deeply, studied. This article reviews these effects on whole animals, animal cells, and microorganisms. Both "solvent isotope effects," those due to the special properties of D20 as a solvent, and "deuterium isotope effects" (DIE), which result when D replaces H in many biological molecules, are considered. The low toxicity of D20 toward mammals is reflected in its widespread use for measuring water spaces in humans and other animals. Higher concentrations (usually >20% of body weight) can be toxic to animals and animal cells. Effects on the nervous system and the liver and on formation of different blood cells have been noted. At the cellular level, D20 may affect mitosis and membrane function. Protozoa are able to withstand up to 70% D20. Algae and bacteria can adapt to grow in 100% D2O and can serve as sources of a large number of deuterated molecules. D2O increases heat stability of macromolecules but may decrease cellular heat stability, possibly as a result of inhibition of chaperonin formation. High D2O concentrations can reduce salt- and ethanol-induced hypertension in rats and protect mice from gamma irradation. Such concentrations are also used in boron neutron capture therapy to increase neutron penetration to boron compounds bound to malignant cells. D2O is more toxic to malignant than normal animal cells, but at concentrations too high for regular therapeutic use. D2O and deuterated drugs are widely used in studies of metabolism of drugs and toxic substances in humans and other animals. The deuterated forms of drugs often have different actions than the protonated forms. Some deuterated drugs show different transport processes. Most are more resistant to metabolic changes, especially those changes mediated by cytochrome P450 systems. Deuteration may also change the pathway of drug metabolism (metabolic switching). Changed metabolism may lead to increased duration of action and lower toxicity. It may also lead to lower activity, if the drug is normally changed to the active form in vivo. Deuteration can also lower the genotoxicity of the anticancer drug tamoxifen and other compounds. Deuteration increases effectiveness of long-chain fatty acids and fluoro-D-phenylalanine by preventing their breakdown by target microorganisms. A few deuterated antibiotics have been prepared, and their antimicrobial activity was found to be little changed. Their action on resistant bacteria has not been studied, but there is no reason to believe that they would be more effective against such bacteria. Insect resistance to insecticides is very often due to insecticide destruction through the cytochrome P450 system. Deuterated insecticides might well be more effective against resistant insects, but this potentially valuable possibility has not yet been studied.

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“…This is of significant importance as it means that there are no changes in the key metabolic pathway products. Eukaryotic microorganisms do not suffer the same level of deuterium inhibition as prokaryotes although adaptation may still be necessary to achieve effective growth (Haon et al 1993;Morgan et al 2000). Kushner et al (1997) stated 'mechanisms involved in such adaptation have hardly been studied' and the situation appears to be largely unchanged at the present time.…”

Section: Discussionmentioning

confidence: 99%

Directed microbial biosynthesis of deuterated biosurfactants and potential future application to other bioactive molecules

Smyth

Perfumo

Marchant

et al. 2010

Appl Microbiol Biotechnol

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Deuterated rhamnolipids were produced using strain AD7 of Pseudomonas aeruginosa, which was progressively adapted to increasing levels of deuterium in D(2)O and carbon substrates. Fourteen different deuterated rhamnolipid structures, including structural isomers, were produced which is similar to normal protonated structures. There were two main products monorhamnolipid Rha-C(10)-C(10) and dirhamnolipid Rha(2)-C(10)-C(10). The levels of deuteration varied from 16% with 25% D(2)O + h-glycerol to 90% with 100% D(2)O + d-glycerol. When d-tetradecane was used with H(2)O, virtually all the deuterium appeared in the lipid chains while using h-tetradecane + D(2)O led to the majority of deuterium in the sugars. The adaptation to growth in deuterium appeared to be metabolic since no genetic changes could be found in the key rhamnolipid biosynthetic genes, the rhamnosyl transferases RhlB and RhlC. Deuterated sophorolipids were similarly produced using Candida bombicola and Candida apicola although in this case, no adaptation process was necessary. Up to 40 different sophorolipids were produced by these yeasts. However, unlike the rhamnolipids, use of D(2)O did not lead to any deuteration of the lipid chains, but direct incorporation into the lipid was achieved using d-isostearic acid. The results from these experiments show the feasibility of producing deuterated bioactive compounds from microorganisms coupled with the possibility of manipulating the pattern of labelling through judicious use of different deuterated substrates.

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Low cost production of perdeuterated biomass using methylotrophic yeasts

Cited by 16 publications

References 25 publications

Expression and pharmacological characterization of the human μ‐opioid receptor in the methylotrophic yeast Pichia pastoris

Expression and pharmacological characterization of the human μ‐opioid receptor in the methylotrophic yeast Pichia pastoris

Pharmacological uses and perspectives of heavy water and deuterated compounds

Directed microbial biosynthesis of deuterated biosurfactants and potential future application to other bioactive molecules

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