Improved hydrogen production with expression of hemH and lba genes in chloroplast of Chlamydomonas reinhardtii

Wu, Shuangxiu; Huang, Rui; Xu, Lili; Yang, Guangyu; Wang, Quanxi

doi:10.1016/j.jbiotec.2010.01.023

Cited by 39 publications

(20 citation statements)

References 26 publications

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“…However, most of the recombinant proteins produced, have been soluble proteins, and furthermore have generally been proteins that can be considered 'benign' in that they have no effect on chloroplast metabolism. To-date there has been only a few reported attempts to express membrane-associated proteins, or enzymes that introduce novel metabolic pathways (Blatti et al, 2012;Wu et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

et al. 2015

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The unicellular green alga Chlamydomonas reinhardtii has potential as a cell factory for the production of recombinant proteins and other novel compounds, but mainstream adoption has been hindered by a scarcity of genetic tools and a need to identify products that can be generated in a cost-effective manner. A promising strategy is to use algal chloroplasts as a site for synthesis of high value bioactive compounds such as diterpenoids since these are derived from metabolic building blocks that occur naturally within the organelle. However, synthesis of these complex plant metabolites requires the introduction of membraneassociated enzymes including cytochrome P450 enzymes (P450s). Here, we show that a gene (CYP79A1) encoding a model P450 can be introduced into the C. reinhardtii chloroplast genome using a simple transformation system. The gene is stably expressed and the P450 is efficiently targeted into chloroplast membranes, where it is active and accounts for 0.4% of total cell protein. These results provide proof of concept for the introduction of diterpenoid synthesis pathways into the chloroplast of Chlamydomonas reinhardtii.

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Section: Introductionmentioning

confidence: 99%

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

et al. 2015

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“…Because of their importance in ecology and industry, algae are now considered promising organisms for economical and industrial applications and are thus a target of genetic transformation (Walker et al, 2005;Hallmann, 2007;Blouin et al, 2011). To date, genetic transformation has succeeded in microalgae; thus, stably transformed microalgae are now employed to produce recombinant antibodies, vaccines, or bio-hydrogen as well as to analyze the gene functions targeted for engineering (Sun et al, 2003;Zorin et al, 2009;Specht et al, 2010;Wu et al, 2010). However, it has proven difficult to establish transgenic macroalgae, which has hampered understanding their gene functions in various physiological regulations and also their utilization in biotechnological applications.…”

Section: Introductionmentioning

confidence: 99%

Transient Transformation of Red Algal Cells: Breakthrough Toward Genetic Transformation of Marine Crop Porphyra Species

Mikami¹,

Hirata²,

Takahashi³

et al. 2011

Genetic Transformation

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“…By transferring the coding region of both the ferrochelatase gene, hemH, from Bradyrhizobium japonicum, and the leghemoglobin gene, lba, from Glycine max, into the chloroplast of C. reinhardtii, transgenic strains that accumulate leghemoglobine in the chloroplast were created. These strains more rapidly consumed O 2 and displayed increased H 2 production in S-free medium (Wu et al 2010). This result demonstrates the potential of local buffering of O 2 in the chloroplast in order to help overcoming the incompatibility of oxygenic photosynthesis and H 2 photoproduction and opens new possibilities for the development of more efficient photoautotrophic H 2 production protocols in S-deplete or even S-replete conditions.…”

Section: Elimination Of Electron Sinks Is a Prerequisite For Sustainementioning

confidence: 74%

“…However, higher respiration rates would increase mitochondrial consumption of reducing equivalents derived from C-metabolism and would therefore limit the amount of reducing equivalents available for supporting H 2 evolution through the indirect PSII-independent pathway. Wu et al (2010) showed that O 2 evolved from PSII activity can also be trapped without consumption of reducing equivalents. In this study, the dependence of the anoxic conditions on mitochondrial consumption could be partly circumvented by local buffering of the evolved O 2 with O 2 trapping proteins.…”

Section: Elimination Of Electron Sinks Is a Prerequisite For Sustainementioning

confidence: 99%

Hydrogen photo-evolution upon S deprivation stepwise: an illustration of microalgal photosynthetic and metabolic flexibility and a step stone for future biotechnological methods of renewable H2 production

Ghysels

Franck

2010

Photosynth Res

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The metabolic flexibility of some photosynthetic microalgae enables them to survive periods of anaerobiosis in the light by developing a particular photofermentative metabolism. The latter entails compounds of the photosynthetic electron transfer chain and an oxygen-sensitive hydrogenase in order to reoxidize reducing equivalents and to generate ATP for maintaining basal metabolic function. This pathway results in the photo-evolution of hydrogen gas by the algae. A decade ago, Melis and coworkers managed to reproduce such a condition in a laboratory context by depletion of sulfur in the algal culture media, making the photo-evolution by the algae sustainable for several days (Melis et al. in Plant Physiol 122:127-136, 2000). This observation boosted research in algal H(2) evolution. A feature, which due to its transient nature was long time considered as a curiosity of algal photosynthesis suddenly became a phenomenon with biotechnological potential. Although the Melis procedure has not been developed into a biotechnological process of renewable H(2) generation so far, it has been a useful tool for studying microalgal metabolic and photosynthetic flexibility and a possible step stone for future H(2) production procedures. Ten years later most of the critical steps and limitations of H(2) production by this protocol have been studied from different angles particularly with the model organism Chlamydomonas reinhardtii, by introducing various changes in culture conditions and making use of mutants issued from different screens or by reverse genomic approaches. A synthesis of these observations with the most important conclusions driven from recent studies will be presented in this review.

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Improved hydrogen production with expression of hemH and lba genes in chloroplast of Chlamydomonas reinhardtii

Cited by 39 publications

References 26 publications

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

Transient Transformation of Red Algal Cells: Breakthrough Toward Genetic Transformation of Marine Crop Porphyra Species

Hydrogen photo-evolution upon S deprivation stepwise: an illustration of microalgal photosynthetic and metabolic flexibility and a step stone for future biotechnological methods of renewable H2 production

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