Production of mouse interleukin‐12 is greater in tobacco hairy roots grown in a mist reactor than in an airlift reactor

Liu, Chun-Zhao; Towler, Melissa J.; Medrano, Giuliana; Cramer, Carole L.; Weathers, Pamela J.

doi:10.1002/bit.22154

Cited by 65 publications

(21 citation statements)

References 47 publications

(53 reference statements)

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“…This method has been normally used in a trickle bed bioreactor (Grzegorczyk and Wysokinska 2010). The various mist bioreactor configurations used earlier for A. annua hairy roots were acoustic window mist reactor (Souret et al 2003;Towler et al 2006, submerged ultrasonic bioreactor (Dilorio et al 1992;Liu et al 1999) and sonic nozzle in mist reactor (Liu et al 2009). The less biomass and artemisinin concentration obtained in a conventional nutrient mist (Table 1) could be attributed to starvation of hairy roots due to an inadequate supply of nutrients through mist.…”

Section: Discussionmentioning

confidence: 99%

Artemisinin production by plant hairy root cultures in gas- and liquid-phase bioreactors

Patra

Srivastava

2015

Plant Cell Rep

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Alternative biotechnological protocol for large-scale artemisinin production was established. It featured enhanced growth and artemisinin production by cultivation of hairy roots in nutrient mist bioreactor (NMB) coupled with novel cultivation strategies. Artemisinin is used for the treatment of cerebral malaria. Presently, its main source is from seasonal plant Artemisia annua. This study featured investigation of growth and artemisinin production by A. annua hairy roots (induced by Agrobacterium rhizogenes-mediated genetic transformation of explants) in three bioreactor configurations-bubble column reactor, NMB and modified NMB particularly to establish their suitability for commercial production. It was observed that cultivation of hairy roots in a non-stirred bubble column reactor exhibited a biomass accumulation of 5.68 g/l only while batch cultivation in a custom-made NMB exhibited a higher biomass concentration of 8.52 g/l but relatively lower artemisinin accumulation of 0.22 mg/g was observed in this reactor. A mixture of submerged liquid-phase growth (for 5 days) followed by gas-phase cultivation in nutrient mist reactor operation strategy (for next 15 days) was adopted for hairy root cultivation in this investigation. Reasonably, high (23.02 g/l) final dry weight along with the artemisinin accumulation (1.12 mg/g, equivalent to 25.78 mg/l artemisinin) was obtained in this bioreactor, which is the highest reported artemisinin yield in the gas-phase NMB cultivation.

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Section: Discussionmentioning

confidence: 99%

Artemisinin production by plant hairy root cultures in gas- and liquid-phase bioreactors

Patra

Srivastava

2015

Plant Cell Rep

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“…This may fairly help in filling the gap between capital cost and the benefits of technology at industrial scale (Stiles and Liu 2013). Recently, hairy root cultures are also used for bioremediation of toxic compounds and the production of recombinant proteins at large scale (Liu et al 2009;Sosa Alderete et al 2012). Thus, a combination of bioreactor technology with other hairy root-based research applications will definitely extract scientific and technological solutions to the important socioeconomic issues like environment and health.…”

Section: Bioreactor Technologymentioning

confidence: 98%

Hairy root biotechnology—indicative timeline to understand missing links and future outlook

et al. 2015

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Agrobacterium rhizogenes-mediated hairy roots (HR) were developed in the laboratory to mimic the natural phenomenon of bacterial gene transfer and occurrence of disease syndrome. The timeline analysis revealed that during 90 s, the research expanded to the hairy root-based secondary metabolite production and different yield enhancement strategies like media optimization, up-scaling, metabolic engineering etc. An outlook indicates that much emphasis has been given to the strategies that are helpful in making this technology more practical in terms of high productivity at low cost. However, a sequential analysis of literature shows that this technique is upgraded to a biotechnology platform where different intra- and interdisciplinary work areas were established, progressed, and diverged to provide scientific benefits of various hairy root-based applications like phytoremediation, molecular farming, biotransformation, etc. In the present scenario, this biotechnology research platform includes (a) elemental research like hairy root-mediated secondary metabolite production coupled with productivity enhancement strategies and (b) HR-based functional research. The latter comprised of hairy root-based applied aspects such as generation of agro-economical traits in plants, production of high value as well as less hazardous molecules through biotransformation/farming and remediation, respectively. This review presents an indicative timeline portrayal of hairy root research reflected by a chronology of research outputs. The timeline also reveals a progressive trend in the state-of-art global advances in hairy root biotechnology. Furthermore, the review also discusses ideas to explore missing links and to deal with the challenges in future progression and prospects of research in all related fields of this important area of plant biotechnology.

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“…They also offer advantages for production of engineered proteins (Guillon et al, 2006; Liu et al, 2009; Shanks and Morgan, 1999; Zhang et al, 2005). A real bottleneck, however, exists in the production of useful compounds from hairy roots, especially small molecules, for two main reasons: (1) many of the relevant biochemical pathways and their regulation are inadequately understood; and (2) cost-effective scalable production systems are not yet well developed.…”

Section: Introductionmentioning

confidence: 99%

“…This poses a challenge to reactor scale-up because the inoculum levels required to create a dense bed in a reactor quickly become impractical as the reactor size increases. To address this challenge we incorporated a flexible wall growth chamber (a constricted plastic bag) into the mist reactor in order to increase the apparent density of the inoculum root bed early in the growth of the culture to facilitate nutrient droplet capture (Liu et al, 2009). As the roots grow, they can expand the bag and thus maintain a reasonable bed density.…”

Section: Introductionmentioning

confidence: 99%

“…Although the misting head cannot be autoclaved, it can be sterilized using ethylene oxide. We have also developed several methods for sterilization that can be used to prevent contamination in the reactor (Liu et al, 2009; Sharaf-Eldin and Weathers, 2006). The plastic bag growth chamber and the durable high-throughput sonic mister now make the mist reactor quite scalable, easy to use, and when scaled to 20 L, low in capital costs.…”

Section: Introductionmentioning

confidence: 99%

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Biomass production of hairy roots of Artemisia annua and Arachis hypogaea in a scaled‐up mist bioreactor

Sivakumar¹,

Liu²,

Towler³

et al. 2010

Biotech & Bioengineering

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Hairy roots have the potential to produce a variety of valuable small and large molecules. The mist reactor is a gas phase bioreactor that has shown promise for low-cost culture of hairy roots. Using a newer, disposable culture bag, mist reactor performance was studied with two species, Artemisia annua L. and Arachis hypogaea (peanut), at scales from 1 to 20 L. Both species of hairy roots when grown at 1 L in the mist reactor showed growth rates that surpassed that in shake flasks. From the information gleaned at 1 L, Arachis was scaled further to 4 and then 20 L. Misting duty cycle, culture medium flow rate, and timing of when flow rate was increased were varied. In a mist reactor increasing the misting cycle or increasing the medium flow rate are the two alternatives for increased delivery of liquid nutrients to the root bed. Longer misting cycles beyond 2–3 min were generally deemed detrimental to growth. On the other hand, increasing the medium flow rate to the sonic nozzle especially during the exponential phase of root growth (weeks 2–3) was the most important factor for increasing growth rates and biomass yields in the 20 L reactors. A. hypogaea growth in 1 L reactors was μ = 0.173 day−1 with biomass yield of 12.75 g DWL−1. This exceeded that in shake flasks at μ = 0.166 day−1 and 11.10 g DWL−1. Best growth rate and biomass yield at 20 L was μ = 0.147 and 7.77 g DWL−1, which was mainly achieved when medium flow rate delivery was increased. The mist deposition model was further evaluated using this newer reactor design and when the apparent thickness of roots (+hairs) was taken into account, the empirical data correlated with model predictions. Together these results establish the most important conditions to explore for future optimization of the mist bioreactor for culture of hairy roots.

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Production of mouse interleukin‐12 is greater in tobacco hairy roots grown in a mist reactor than in an airlift reactor

Cited by 65 publications

References 47 publications

Artemisinin production by plant hairy root cultures in gas- and liquid-phase bioreactors

Artemisinin production by plant hairy root cultures in gas- and liquid-phase bioreactors

Hairy root biotechnology—indicative timeline to understand missing links and future outlook

Biomass production of hairy roots of Artemisia annua and Arachis hypogaea in a scaled‐up mist bioreactor

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