Coexpression of redox partners increases the hydrocortisone (cortisol) production efficiency in CYP11B1 expressing fission yeast Schizosaccharomyces pombe

Hakki, Tarek; Zearo, Silvia; Drăgan, Călin-Aurel; Bureik, Matthias; Bernhardt, Rita

doi:10.1016/j.jbiotec.2007.06.022

Cited by 40 publications

(24 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The production of hydrocortisone in this recombinant fission yeast was considerably higher than in S. cerevisiae expressing bovine CYP11B1. Recently, the 11␤-hydroxylation of 11-deoxycortisol in fission yeast was further improved by coexpression of the natural redox partners of CYP11B1, adrenodoxin and its reductase (Hakki et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Discovery of a steroid 11α-hydroxylase from Rhizopus oryzae and its biotechnological application

Petrič

Hakki

Bernhardt

et al. 2010

Journal of Biotechnology

View full text Add to dashboard Cite

Section: Introductionmentioning

confidence: 99%

Discovery of a steroid 11α-hydroxylase from Rhizopus oryzae and its biotechnological application

Petrič

Hakki

Bernhardt

et al. 2010

Journal of Biotechnology

View full text Add to dashboard Cite

“…The same observation was made when the cells were incubated with the CYP11B1 substrate RSS (or solvent alone). Recently, it was found that AdR coexpression moderately increases steroid hydroxylation activity of fission yeast strains expressing CYP11B1 and Adx [28], but not of strains expressing CYP11B2 and Adx (Hakki, unpublished observations), indicating that both expression level and activity of arh1 (syn. SPBC3B8.01c, the S. pombe homologue of AdR) are sufficient for reduction of most of the overexpressed Adx.…”

Section: Resultsmentioning

confidence: 99%

“…Moreover, its resistance to ROS is still another property that renders fission yeast a particularly suited model organism for the recombinant expression of mammalian cytochromes P450 for biotechnological applications, like production of hydroxysteroids [27,28,33] or testing of cytochrome P450 inhibitors [34,35]. Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Expression of CYP11B1 in strain SZ1 and its mitochondrial localization have been described [27]. Plasmid pTH2 contains two separate expression cassettes for human AdR (including its original mitochondrial targeting sequence) and bovine Adx-WT (fused to the CYP11B1 localization sequence as above), respectively, both under control of the nmt1 promotor; mitochondrial localization of AdR expressed from this plasmid has been shown previously [28].…”

Section: Expression Vector and Cdnamentioning

confidence: 92%

See 1 more Smart Citation

ROS production by adrenodoxin does not cause apoptosis in fission yeast

et al. 2007

Self Cite

View full text Add to dashboard Cite

We previously showed that production of reactive oxygen species (ROS) caused by overexpression of the mitochondrial electron transfer protein adrenodoxin (Adx) induces apoptosis in mammalian cells. In the fission yeast Schizosaccharomyces pombe, ROS are also produced in cells that undergo an apoptotic-like cell death, but it is not yet clear whether they are actually causative for this phenomenon or whether they are merely produced as a by-product. Therefore, the purpose of this study was to trigger mitochondrial ROS production in fission yeast by overexpression of either wildtype Adx (Adx-WT) or of several activated Adx mutants and to investigate its consequences. It was found that strong expression of either Adx-WT or Adx-S112W did not produce any ROS, while Adx-D113Y caused a twofold and Adx1-108 a threefold increase in ROS formation as compared to basal levels. However, no typical apoptotic markers or decreased viability could be observed in these strains. Since we previously observed that an increase in mitochondrial ROS formation of about 60% above basal levels is sufficient to strongly induce apoptosis in mammalian cells, we conclude that S. pombe is either very robust to mitochondrial ROS production or does not undergo apoptotic cell death in response to mitochondrial ROS at all.

show abstract

“…The choice of heterologous redox partners is firstly determined by the class of P450 but has proven not to be that straight forward, and optimization of the electron transfer has been shown to be beneficial, e.g., by a modified ferredoxin ). Putidaredoxin reductase with putidaredoxin (Kim and Ortiz de Montellano 2009) and adrenodoxin reductase with adrenodoxin (Zehentgruber et al 2010a;Hollmann et al 2006;Ewen et al 2012;Hakki et al 2008) have been the most commonly used redox partners for heterologous expression. Non-enzymatic transfer of electrons to the heme-active site has also been explored, including direct chemical reduction or electrochemical reduction (Hlavica 2009) as well as light driven catalysis exemplified by hydroxylation of lauric acid by P450 BM3 (Tran et al 2013).…”

Section: Electron Transport and Coupling Efficiencymentioning

confidence: 99%

Guidelines for development and implementation of biocatalytic P450 processes

Lundemo

Woodley

2015

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Biocatalytic reactions performed by cytochrome P450 monooxygenases are interesting in pharmaceutical research since they are involved in human drug metabolism. Furthermore, they are potentially interesting as biocatalysts for synthetic chemistry because of the exquisite selectivity of the chemistry they undertake. For example, selective hydroxylation can be undertaken on a highly functionalized molecule without the need for functional group protection. Recent progress in the discovery of novel P450s as well as protein engineering of these enzymes strongly encourages further development of their application, including use in synthetic processes. The biological characteristics of P450s (e.g., cofactor dependence) motivate the use of whole-cell systems for synthetic processes, and those processes implemented in industry are so far dominated by growing cells and native host systems. However, for an economically feasible process, the expression of P450 systems in a heterologous host with sufficient biocatalyst yield (g/g cdw) for non-growing systems or space-time yield (g/L/h) for growing systems remains a major challenge. This review summarizes the opportunities to improve P450 whole-cell processes and strategies in order to apply and implement them in industrial processes, both from a biological and process perspective. Indeed, a combined approach of host selection and cell engineering, integrated with process engineering, is suggested as the most effective route to implementation.

show abstract

Coexpression of redox partners increases the hydrocortisone (cortisol) production efficiency in CYP11B1 expressing fission yeast Schizosaccharomyces pombe

Cited by 40 publications

References 26 publications

Discovery of a steroid 11α-hydroxylase from Rhizopus oryzae and its biotechnological application

Discovery of a steroid 11α-hydroxylase from Rhizopus oryzae and its biotechnological application

ROS production by adrenodoxin does not cause apoptosis in fission yeast

Guidelines for development and implementation of biocatalytic P450 processes

Contact Info

Product

Resources

About