LEDitSHAKE: a lighting system to optimize the secondary metabolite content of plant cell suspension cultures

Abstract

Plant secondary metabolites are widely used in the food, cosmetic and pharmaceutical industries. They can be extracted from sterile grown plant cell suspension cultures, but yields and quality strongly depend on the cultivation environment, including optimal illumination. Current shaking incubators do not allow different light wavelengths, intensities and photoperiods to be tested in parallel. We therefore developed LEDitSHAKE, a system for multiplexed customized illumination within a single shaking incubator. We used 3D printing to integrate light-emitting diode assemblies into flask housings, allowing 12 different lighting conditions (spectrum, intensity and photoperiod) to be tested simultaneously. We did a proof of principle of LEDitSHAKE using the system to optimize anthocyanin production in grapevine cell suspension cultures. The effect of 24 different light compositions on the total anthocyanin content of grapevine cell suspension cultures was determined using a Design of Experiments approach. We predicted the optimal lighting conditions for the upregulation and downregulation of 30 anthocyanins and found that short-wavelength light (blue, UV) maximized the concentration of most anthocyanins, whereas long-wavelength light (red) had the opposite effect. Therefore our results demonstrate proof of principle that the LEDitSHAKE system is suitable for the optimization of processes based on plant cell suspension cultures.

Reducing water uptake into BY‐2 cells by systematically optimizing the cultivation parameters increases product yields achieved by transient expression in plant cell packs

“…BY‐2 cells were cultivated at 26°C, shaking at 160 rpm and continuously exposed to light during the experiment (24 h d ‐1 ) using a LEDitSHAKE device. [ 25 ] In all experiments, BY‐2 cells were harvested after 168 h and PCPs were incubated for 72 h post‐infiltration. Conditions representing “white light” (dashed red line) were simulated as mixtures of blue, green, red, and far‐red light (equal intensity).…”

“…BY‐2 cells were cultivated in 5.0‐L stirred‐tank bioreactors (100 g wet biomass L ‐1 , packed cell volume 30% to 40% [v v ‐1 ]) as previously described, [ 15 ] in 0.25‐L glass flasks containing 0.05 L medium (routine cultivations), or in 0.10‐L glass flasks containing 0.02 L medium housed in a LEDitSHAKE device. [ 25 ] Cells were cultivated at 26°C, ∼ 50% relative humidity, shaking orbitally at 160 rpm with an excentricity of 50 mm in a Climo‐Shaker ISF1‐X (Kühner, Birsfelden, Germany). Shake‐flask cultures intended for expression studies were inoculated with a seeding cell density of 2 × 10 8 cells L ‐1 and were harvested after 5 to 8 days.…”

“…BY‐2 cells were also exposed to artificial white light (Osram Lumilux Cool White, HO 49W/840; Figure S1) and optionally to defined wavelengths (Table S3), including far‐red (peak = 730 nm), red (peak = 660 nm), green (peak = 505 nm) and blue (peak = 455 nm) light, as well as mixtures thereof, in the LEDitSHAKE device. [ 25 ] The total light intensity in these experiments was set to 20 to 40 µmol m ‐2 s ‐1 and cells were continuously exposed to light (24 h ‐1 d ‐1 ) for 3 weeks.…”

Trends in the Tissue Culture Techniques and the Synthesis of Bioactive Compounds in Eurycoma longifolia Jack—Current Status and Future Perspectives

“…Different LED monochromatic lights have been employed individually or in combinations to enhance the synthesis of crucial metabolites. LEDs have the ability to improve metabolite production in both callus [ 118 , 119 , 120 , 121 , 122 ] and cell suspension [ 123 , 124 ]. Moreover, several studies highlight the effectiveness of LEDs in enhancing the synthesis of bioactive compounds in various in vitro cultures [ 116 ].…”

Effect assessment for the interaction between shaking table and eccentric load

“…Electro-hydraulic shaking table is one of the most important pieces of equipment for replicating actual vibration conditions in various applications, including biomedical and geological researches 1 – 3 , engineering seismic research works 4 – 6 , aerospace vibration tests 7 , 8 , and vehicle road simulations 9 , 10 , etc. In the context of shaking table test, the control-structure interaction (CSI) 11 – 13 is the dynamic coupling between shaking table and load (that is the test structure model) (Fig.…”