Process engineering of biopharmaceutical production in moss bioreactors via model-based description and evaluation of phytohormone impact

The moss Physcomitrella is an interesting production host for recombinant biopharmaceuticals. Here we produced MFHR1, a synthetic complement regulator which has been proposed for the treatment of diseases associated to the complement system as part of human innate immunity. We studied the impact of different operation modes for the production process in 5 L stirred-tank photobioreactors. The total amount of recombinant protein was doubled by using fed-batch or batch compared to semi-continuous operation, although the maximum specific productivity (mg MFHR1/g FW) increased just by 35%. We proposed an unstructured kinetic model which fits accurately with the experimental data in batch and semi-continuous operation under autotrophic conditions with 2% CO2 enrichment. The model is able to predict recombinant protein production, nitrate uptake and biomass growth, which is useful for process control and optimization. We investigated strategies to further increase MFHR1 production. While mixotrophic and heterotrophic conditions decreased the MFHR1-specific productivity compared to autotrophic conditions, addition of the phytohormone auxin (NAA, 10 μM) to the medium enhanced it by 470% in shaken flasks and up to 230% and 260%, in batch and fed-batch bioreactors, respectively. Supporting this finding, the auxin-synthesis inhibitor L-Kynurenine (100 μM) decreased MFHR1 production significantly by 110% and 580% at day 7 and 18, respectively. Expression analysis revealed that the MFHR1 transgene, driven by the Physcomitrella actin5 (PpAct5) promoter, was upregulated 16 hours after NAA addition and remained enhanced over the whole process, whereas the auxin-responsive gene PpIAA1A was upregulated within the first two hours, indicating that the effect of auxin on PpAct5 promoter-driven expression is indirect. Furthermore, the day of NAA supplementation was crucial, leading to an up to 8-fold increase of MFHR1-specific productivity (0.82 mg MFHR1/ g fresh weight, 150 mg accumulated over 7 days) compared to the productivity reported previously. Our findings are likely to be applicable to other plant-based expression systems to increase biopharmaceutical production and yields.

Light-Mediated Reduction in Photosynthesis in Closed Greenhouses Can Be Compensated for by CO2 Enrichment in Tomato Production

Concepts of semi-closed greenhouses can be used to save energy, whereas their technical equipment often causes a decrease in the light received by the plants. Nevertheless, higher yields are achieved, which are presumably triggered by a higher CO2 concentration in the greenhouse and associated higher photosynthesis because of the technical cooling and the longer period of closed ventilation. Therefore, we examined the effects of photosynthetic photon flux density (PPFD) and CO2 concentration on plant photosynthesis and transpiration in tomato using a multiple cuvette gas exchange system. In a growth chamber experiment, we demonstrated that a light-mediated reduction in photosynthesis can be compensated or even overcompensated for by rising CO2 concentration. Increasing the CO2 concentration from 400 to 1000 µmol mol−1 within the PPFD range from 303 to 653 µmol m−2 s−1 resulted in an increase in net photosynthesis of 51%, a decrease in transpiration of 5 to 8%, and an increase in photosynthetic water use efficiency of 60%. Estimations showed that light reductions of 10% can be compensated for via increasing the CO2 concentration by about 100 µmol mol−1 and overcompensated for by about 40% if CO2 concentration is kept at 1000 instead of 400 µmol mol−1.

Preparation and Properties of CO2 Micro-Nanobubble Water Based on Response Surface Methodology

Carbon dioxide (CO2) enrichment in an agricultural environment has been shown to enhance the efficiency of crop photosynthesis, increasing crop yield and product quality. There is a problem of the excessive use of CO2 gas when the CO2 is enriched for crops, such as soybean and other field crops. Given the application of micro-nanobubbles (MNBs) in agricultural production, this research takes CO2 as the gas source to prepare the micro-nanobubble water by the dissolved gas release method, and the response surface methodology is used to optimize the preparation process. The results show that the optimum parameters, which are the gas–liquid ratio, generator running time, and inlet water temperature for the preparation of CO2 micro-nanobubble water, are 2.87%, 28.47 min, and 25.52 °C, respectively. The CO2 content in the MNB water prepared under the optimum parameters is 7.64 mg/L, and the pH is 4.08. Furthermore, the particle size of the bubbles is mostly 255.5 nm. With the extension of the storage time, some of the bubbles polymerize and spill out, but there is still a certain amount of nanoscale bubbles during a certain period. This research provides a new idea for using MNB technology to increase the content and lifespan of CO2 in water, which will slow the release and increase the utilization of CO2 when using CO2 enrichment in agriculture.

Integrative Assessment of Sediments Affected by CO2 Enrichment: A Case Study in the Bay of Santos—SP, Brazil

CO2 enrichment in the marine environment caused by leakages from carbon capture and storage technologies may occur over operational procedures. An integrated approach using weight-of-evidence was applied to assess the environmental risk associated with the acidification caused by CO2 enrichment in coastal sediments from Santos (Brazil). Chemical analyses (metal(loid)s and organic contaminant (e.g., hydrocarbons), toxicity tests (amphipods mortality, sea-urchin embryo-larval development) and macro-benthic community structure alteration assessment were performed with different acidified scenarios (pH 8.0–6.0) for two stations with different contamination degrees. These lines of evidence were statistically analyzed and integrated (multivariate analysis and ANOVA). Results of toxicity showed significant chronic effects starting at pH 7.0 while acute effects were observed starting at pH 6.5. The macro-benthic community integrity showed significant differences for all treatments at the Piaçaguera channel station, considered to be moderately contaminated. Results from the multivariate analysis correlated toxic effects and increase in the mobility of some elements with acidification. Also, the biological indexes were correlated with concentrations of dissolved Zn in seawater. The pH of 6.0 was extremely toxic for marine life due to its high acidification and metal bioavailability. The approach herein identified and discriminated the origin of the degradation caused by the acidification related to the enrichment of CO2.

Soybean-BioCro: A semi-mechanistic model of soybean growth

Soybean is a major global source of protein and oil. Understanding how soybean crops will respond to the changing climate and identifying the responsible molecular machinery, are important for facilitating bioengineering and breeding to meet the growing global food demand. The BioCro family of crop models are semi-mechanistic models scaling from biochemistry to whole crop growth and yield. BioCro was previously parameterized and proved effective for the biomass crops miscanthus, coppice willow, and Brazilian sugarcane. Here, we present Soybean-BioCro, the first food crop to be parameterized for BioCro. Two new module sets were incorporated into the BioCro framework describing the rate of soybean development and carbon partitioning and senescence. The model was parameterized using field measurements collected over the 2002 and 2005 growing seasons at the open air [CO2] enrichment (SoyFACE) facility under ambient atmospheric [CO2]. We demonstrate that Soybean-BioCro successfully predicted how elevated [CO2] impacted field-grown soybean growth without a need for re-parameterization, by predicting soybean growth under elevated atmospheric [CO2] during the 2002 and 2005 growing seasons, and under both ambient and elevated [CO2] for the 2004 and 2006 growing seasons. Soybean-BioCro provides a useful foundational framework for incorporating additional primary and secondary metabolic processes or gene regulatory mechanisms that can further aid our understanding of how future soybean growth will be impacted by climate change.

Effect of elevated CO2 and elevated temperature on growth and biomass accumulation in Valeriana jatamansi Jones. under different nutrient status in the western Himalaya

Valeriana jatamansi is an important medicinal and aromatic plant used as sedative in modern and traditional medicines butthere is dearth of literature regarding how elevated CO2 and temperature affect on this plant. Therefore,an experiment was conducted to study the effect of elevated CO2 (550±50 µmol mol-1) and elevated temperature (2.5±0.5°C above ambient) and vermicompost on growth, phenology and biomass accumulation in V. jatamansi under Free Air CO2 Enrichment (FACE) and Free Air TemperatureIncrement (FATI) facilities at Palampur, India, during 2013-2015. Growth parameters and biomass accumulation into different parts were observed at 4, 12 and 16 months after exposure (MAE). Plant height, total dry biomass and leaf area plant -1 increased in elevated CO2 treatment applied with vermicompost as compared to the other treatments. Elevated CO2 significantly enhanced leaf area (3.5-23.5%), leaf biomass (12.7-33.2%), stem (15.3-15.6%), root (3.2-72.5%), rhizome (2.1-42.2%) and total biomass (7.7-52.7%), whereas elevated temperature increased aboveground biomass (15.0-45.3%), belowground biomass (11.6-55.5%) and total biomass (12.4-7.9%), respectively, as compared to ambient. Phenological stages were advanced by 1.2-3.9 days under FACE and FATI as compared to ambient. The results indicate that aboveground, belowground and total biomass increased under elevated CO2 and elevated temperature as compared to ambient.

Effect of Elevated CO2 on Biomolecules’ Accumulation in Caraway (Carum carvi L.) Plants at Different Developmental Stages

Caraway plants have been known as a rich source of phytochemicals, such as flavonoids, monoterpenoid glucosides and alkaloids. In this regard, the application of elevated CO2 (eCO2) as a bio-enhancer for increasing plant growth and phytochemical content has been the focus of many studies; however, the interaction between eCO2 and plants at different developmental stages has not been extensively explored. Thus, the present study aimed at investigating the changes in growth, photosynthesis and phytochemicals of caraway plants at two developmental stages (sprouts and mature tissues) under control and increased CO2 conditions (ambient CO2 (a CO2, 400 ± 27 μmol CO2 mol−1 air) and eCO2, 620 ± 42 μmol CO2 mol−1 air ppm). Moreover, we evaluated the impact of eCO2-induced changes in plant metabolites on the antioxidant and antibacterial activities of caraway sprouts and mature plants. CO2 enrichment increased photosynthesis and biomass accumulation of both caraway stages. Regarding their phytochemical contents, caraway plants interacted differently with eCO2, depending on their developmental stages. High levels of CO2 enhanced the production of total nutrients, i.e., carbohydrates, proteins, fats and crude fibers, as well as organic and amino acids, in an equal pattern in both caraway sprouts and mature plants. Interestingly, the eCO2-induced effect on minerals, vitamins and phenolics was more pronounced in caraway sprouts than the mature tissues. Furthermore, the antioxidant and antibacterial activities of caraway plants were enhanced under eCO2 treatment, particularly at the mature stage. Overall, eCO2 provoked changes in the phytochemical contents of caraway plants, particularly at the sprouting stage and, hence, improved their nutritive and health-promoting properties.