Jianke Huang

Novel flat-plate photobioreactors for microalgae cultivation with special mixers to promote mixing along the light gradient

Novel flat-plate photobioreactors (PBRs) with special mixers (type-a, type-b, and type-c) were designed based on increased mixing degree along the light gradient. The hydrodynamic and light regime characteristic of the novel PBRs were investigated through computational fluid dynamics. Compared with the control reactor without mixer, the novel reactors can effectively increase liquid velocity along the light gradient, the frequency of light/dark (L/D) cycles, and the algal growth rates of Chlorella pyrenoidosa. The maximum biomass concentrations in type-a, type-b, and type-c reactors were 42.9% (1.3 g L(-1)), 31.9% (1.2 g L(-1)), and 20.9% (1.1 g L(-1)) higher than that in the control reactor (0.91 g L(-1)), respectively, at an aeration rate of 1.0 vvm. Correlation analysis of algal growth rate with the characteristics of mixing and light regime shows the key factors affecting algal photoautotrophic growth are liquid velocity along the light gradient and L/D cycles rather than the macro-mixing degree.

Enhanced lipid productivity of Chlorella pyrenoidosa through the culture strategy of semi-continuous cultivation with nitrogen limitation and pH control by CO2.

Microalgae cultivation with high lipid productivity has received much attention in recent years owing to the economic potential of biofuels production and CO2 emission reduction. Previous studies had reported that the ways of pH-regulation, nutrition-limitation and semi-continuous culture mode can either increase the cells growth rate or promote lipid accumulation. In this study, the novel culture strategy of integrating both nutrition limitation and pH-regulation by CO2 in a semi-continuous cultivation was investigated for enhancing the lipid productivity. In the batch culture, the best growth performance was achieved by controlling pH at 7; lipid contents of the cells can be increased under the nitrogen-limitation conditions. The maximum lipid productivity of 115 mg L(-1) d(-1) was achieved in the novel culture strategy of semi-continuous cultivation with nitrogen-limitation and pH-regulation by CO2, which was 3.64-fold higher than that in the batch culture without pH control and nitrogen limitation.

Genomic Foundation of Starch-to-Lipid Switch in Oleaginous Chlorella spp.

The versatile chlorophyta Chlorella pyrenoidosa provides genomic insights into the trophic diversity and metabolic dynamics. The ability to rapidly switch the intracellular energy storage form from starch to lipids is an advantageous trait for microalgae feedstock. To probe this mechanism, we sequenced the 56.8-Mbp genome of Chlorella pyrenoidosa FACHB-9, an industrial production strain for protein, starch, and lipids. The genome exhibits positive selection and gene family expansion in lipid and carbohydrate metabolism and genes related to cell cycle and stress response. Moreover, 10 lipid metabolism genes might be originated from bacteria via horizontal gene transfer. Transcriptomic dynamics tracked via messenger RNA sequencing over six time points during metabolic switch from starch-rich heterotrophy to lipid-rich photoautotrophy revealed that under heterotrophy, genes most strongly expressed were from the tricarboxylic acid cycle, respiratory chain, oxidative phosphorylation, gluconeogenesis, glyoxylate cycle, and amino acid metabolisms, whereas those most down-regulated were from fatty acid and oxidative pentose phosphate metabolism. The shift from heterotrophy into photoautotrophy highlights up-regulation of genes from carbon fixation, photosynthesis, fatty acid biosynthesis, the oxidative pentose phosphate pathway, and starch catabolism, which resulted in a marked redirection of metabolism, where the primary carbon source of glycine is no longer supplied to cell building blocks by the tricarboxylic acid cycle and gluconeogenesis, whereas carbon skeletons from photosynthesis and starch degradation may be directly channeled into fatty acid and protein biosynthesis. By establishing the first genetic transformation in industrial oleaginous C. pyrenoidosa, we further showed that overexpression of an NAD(H) kinase from Arabidopsis (Arabidopsis thaliana) increased cellular lipid content by 110.4%, yet without reducing growth rate. These findings provide a foundation for exploiting the metabolic switch in microalgae for improved photosynthetic production of food and fuels.

Enhancement of microalgal biomass and lipid productivities by a model of photoautotrophic culture with heterotrophic cells as seed.

For overcoming the long period of seed cultured photoautotrophically and inadequate cell supply for the inoculation of microalgae photoautotrophic cultivation, a model for the photoautotrophic culture of three Chlorella species with heterotrophic cells as seed was investigated. The model can not only take advantages of rapid cell growth in heterotrophic process for preparation of cells as seed but also increase the biomass and lipid productivities of the microalgae cultivated photoautotrophically. The results showed that biomass productivities of Chlorella pyrenoidosa, Chlorella ellipsoidea and Chlorella vulgaris cultured by heterotrophy were 20.9, 26.9 and 25.2 times higher than those by photoautotrophy in seed culturing period. In the subsequent photoautotrophic culture, the biomass and lipid productivities of C. pyrenoidosa, C. ellipsoidea and C. vulgaris with heterotrophic seed were 1.91, 1.51, 1.48 and 1.66, 1.37, 1.42 times higher than those with photoautotrophic seed. Furthermore, the culture model was also carried out successfully outdoor.

Improving performance of flat-plate photobioreactors by installation of novel internal mixers optimized with computational fluid dynamics

A novel mixer was developed to improve the performance of flat-plate photobioreactors (PBRs). The effects of mixer were theoretically evaluated using computational fluid dynamics (CFD) according to radial velocity of fluid and light/dark cycles within reactors. The structure parameters, including the riser width, top clearance, clearance between the baffles and walls, and number of the chambers were further optimized. The microalgae culture test aiming at validating the simulated results was conducted indoor. The results showed the maximum biomass concentrations in the optimized and archetype reactors were 32.8% (0.89 g L(-1)) and 19.4% (0.80 g L(-1)) higher than that in the control reactor (0.67 g L(-1)). Therefore, the novel mixer can significantly increase the fluid velocity along the light attenuation and light/dark cycles, thus further increased the maximum biomass concentration. The PBRs with novel mixers are greatly applicable for high-efficiency cultivation of microalgae.

The effective photoinduction of Haematococcus pluvialis for accumulating astaxanthin with attached cultivation

As the optimal source of astaxanthin, Haematococcus pluvialis was cultured for commercial production of astaxanthin through two continuous phases: cell growth and astaxanthin induction. In this study, the efficiency of an attached system for producing astaxanthin from H. pluvialis was investigated and compared to that of the suspended system (bubble column bioreactor) under various conditions. Results showed that this attached system is more suitable for photoinduction of H. pluvialis than the suspended bioreactor. Under the optimal conditions, the astaxanthin productivity of the attached system was 65.8 mg m(-2)d(-1) and 2.4-fold of that in the suspended system. This attached approach also offers other advantages over suspended systems, such as, producing astaxanthin under a wide range of light intensities and temperatures, saving water, ease to harvest cells, resisting contamination. Therefore, the attached approach can be considered an economical, environmentally friendly and highly-efficient technology for producing astaxanthin from H. pluvialis.

Suppression subtractive hybridization reveals transcript profiling of Chlorella under heterotrophy to photoautotrophy transition.

Background Microalgae have been extensively investigated and exploited because of their competitive nutritive bioproducts and biofuel production ability. Chlorella are green algae that can grow well heterotrophically and photoautotrophically. Previous studies proved that shifting from heterotrophy to photoautotrophy in light-induced environments causes photooxidative damage as well as distinct physiologic features that lead to dynamic changes in Chlorella intracellular components, which have great potential in algal health food and biofuel production. However, the molecular mechanisms underlying the trophic transition remain unclear. Methodology/Principal Findings In this study, suppression subtractive hybridization strategy was employed to screen and characterize genes that are differentially expressed in response to the light-induced shift from heterotrophy to photoautotrophy. Expressed sequence tags (ESTs) were obtained from 770 and 803 randomly selected clones among the forward and reverse libraries, respectively. Sequence analysis identified 544 unique genes in the two libraries. The functional annotation of the assembled unigenes demonstrated that 164 (63.1%) from the forward library and 62 (21.8%) from the reverse showed significant similarities with the sequences in the NCBI non-redundant database. The time-course expression patterns of 38 selected differentially expressed genes further confirmed their responsiveness to a diverse trophic status. The majority of the genes enriched in the subtracted libraries were associated with energy metabolism, amino acid metabolism, protein synthesis, carbohydrate metabolism, and stress defense. Conclusions/Significance The data presented here offer the first insights into the molecular foundation underlying the diverse microalgal trophic niche. In addition, the results can be used as a reference for unraveling candidate genes associated with the transition of Chlorella from heterotrophy to photoautotrophy, which holds great potential for further improving its lipid and nutrient production.

A new technology of CO2 supplementary for microalgae cultivation on large scale - A spraying absorption tower coupled with an outdoor open runway pond.

An effective CO2 supply system of a spraying absorption tower combined with an outdoor ORWP (open raceway pond) for microalgae photoautotrophic cultivation is developed in this paper. The microalgae yield, productivity and CO2 fixation efficiency were investigated, and compared with those of bubbling method. The maximum yield and productivity of biomass were achieved 0.927gL(-1) and 0.114gL(-1)day(-1), respectively. The fixation efficiency of CO2 by microalgae with the spraying tower reached 50%, whereas only 11.17% for bubbling method. Pure CO2 can be used in the spraying absorption tower, and the flow rate was only about one third of the bubbling cultivation. It shows that this new method of quantifiable control CO2 supply can meet the requirements of the growth of microalgae cultivation on large-scale.

Investigation on novel raceway pond with inclined paddle wheels through simulation and microalgae culture experiments

The open raceway ponds are nowadays the most used large-scale reactors for microalgae culture. To avoid the stacking of microalgae, the paddle wheels are the most widely used to circulate and mix the culture medium. In this paper, a numerical simulation using computational fluid dynamics (CFD) was used to investigate the hydrodynamic characteristics of open raceway ponds with different types of paddle wheels (the traditional paddle wheels and the novel paddle wheels with specially inclined angle of the blades). The particle image velocimetry (PIV) was used to validate the reliability of the CFD model. The CFD simulation results showed that the novel raceway pond with 15° inclined angle of the blades had the best mixing efficiency under the same power consumption. Lastly, the results of microalgae culture experiments showed that the growth rates of Chlorella pyrenoidosa in the novel raceway pond with 15° inclined angle of the blades were higher than those in the traditional reactor. The results of the culture experiments and CFD simulations were identical with each other. Therefore, a novel paddle wheel with 15° inclined angle of the blades was obtained for better microalgae cultivation.

Development of a novel multi-column airlift photobioreactor with easy scalability by means of computational fluid dynamics simulations and experiments.

Aiming to culture algae with high efficiency, a novel vertical multi-column airlift photobioreactor (VMAPBR) has been developed. It was constructed with a series of vertically arranged parallel columns with easy scalability. The hydrodynamic, irradiation and shear stress characteristics of the photobioreactor were studied by computational fluid dynamics (CFD). Accordingly, the optimal aeration manner and aeration rate were determined. When the novel airlift PBR was alternately aerated with aeration rate of 0.2vvm, the biomass concentration of Chlorella pyrenoidosa under outdoor condition reached 1.30gL-1 within the prototype PBR and was further increased to 1.56gL-1 within the optimized PBR. The result of cultivation experiment had good agreement with that of CFD prediction.