Guomin Shen

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.

A novel method using CFD to optimize the inner structure parameters of flat photobioreactors

The relationship between growth rates of algae and structure parameters of closed flat-panel photobioreactors was investigated. Ad/Ar (cross-section area of the downcomer/cross-section area of the riser), h0 (clearance from the upper edge of the baffles to the water level), and h1 (clearance from the lower edge of the baffles to the bottom of the reactor) were selected as the inner structure parameters. CFD (Computational Fluid Dynamics) was used to simulate the hydrodynamic parameter (TKE)d (turbulence kinetic energy of the downcomers) and the secondary parameters ε (ratio between td and tc), tc (cycle time), and DZ (dead zones), which were deduced from the hydrodynamic parameters mentioned above. The effects of (TKE)d, ε, tc, DZ, and the inner structure parameters on cell growth of Isochrysis galbana 3011 were analyzed using data collected in 15-L airlift flat-panel photobioreactors. A model was developed to predict algae cell growth based on these inner structure parameters, thereby providing a new method for photobioreactor optimization. Validity of the model was confirmed by experimental data of I. galbana 3011 cultured in 15-L and 300-L photobioreactors, respectively. Finally, the prospect of applying CFD to photobioreactor optimization was discussed.

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.

Changes of biomass, lipid content and fatty acids composition under a light-dark cyclic culture of Chlorella pyrenoidosa in response to different temperature.

For outdoor culture with light-dark cycle, the biomass and lipid losing at night resulted in lowering the biomass and lipid productivity. Previous studies focused on the contents of carbohydrate and protein in response to temperature for production of animal feed and nutritional supplements. In this study, the effects of temperature on the variations of biomass concentration, lipid content and fatty acids composition for production of biofuels were investigated under a light-dark cyclic culture. The results showed that 30 °C was the optimal daytime temperature for achieving high biomass and lipid; raising daytime temperature can lessen night biomass loss and stimulate lipid accumulation. Subsequently, outdoor culture strategy has been improved: keeping culture broth no less than 30 °C during the daytime. Consequently, the net biomass and lipid productivity were increased by 37.8% and 44.9% when compared to the former culture process in the same outdoor climatic conditions.

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.

Medium screening and optimization for photoautotrophic culture of Chlorella pyrenoidosa with high lipid productivity indoors and outdoors

Medium screening and optimization is one of the most important preconditions for photoautotrophic cultivation of microalgae. Although, it has been widely conducted indoors, little work performed outdoors. There are enormous differences between indoor and outdoor conditions, especially for light intensity, temperature and their diurnal or annual fluctuations, which would greatly influence microalgae growth. No data shows whether the differences would lead to different results on medium screening and optimization. In present study, medium screening for the photoautotrophic cultivation of Chlorella pyrenoidosa was carried out indoors and outdoors firstly, and then the selected medium was optimized. The results showed that F-Si medium is the optimum both under indoor and outdoor conditions. Based on F-Si medium, nutrients were optimized as follows: NaNO3 500mgl(-1), NaH2PO4·2H2O 7.7mgl(-1) and FeCl3·6H2O 6.30mgl(-1). With the optimized medium, the biomass, lipid content and productivity were all significantly higher both indoors and outdoors.

Safe antifungal lipopeptides derived from Bacillus marinus B-9987 against grey mold caused by Botrytis cinerea

Abstract Agricultural application studies, including field experiments and acute toxicity tests, were conducted for lipopeptides secreted by marine-derived Bacillus marinus B-9987. Benefiting from commercially available scaled-up lipopeptide purification, the sample of impurities (isolated from target lipopeptides), raw extracted sample (purity: 9.08%), partially purified sample (purity: 20.86%), and highly purified sample (purity: 87.51%) were prepared from B. marinus B-9987 fermentation broth, and used in lab-scale antagonism tests, field experiments, swarming motility tests, and acute toxicity tests. Operations and conditions in field experiments were consistent with the Pesticide-Guidelines for the Field Efficacy Trials (GB/T 17980.28-2000), and acute toxicity tests were executed according to Toxicological Test Methods of Pesticides for Registration (GB 15670-1995). In agar diffusion tests in vitro and pot tests in vivo, all lipopeptide samples with different purities significantly inhibited Botrytis cinerea; meanwhile the sample of impurities isolated from target lipopeptides were not effective against B. cinerea. Results of lab-scale tests showed that the target lipopeptides were effective substances against B. cinerea. Thus, partially purified and raw extracted samples were used in field experiments instead of the highly purified sample for cost saving. In the field experiments against rose grey mold, biological control efficacy of 500 mg L−1 lipopeptides reached 67.53%, slightly lower than 74.05% reached by the agrochemical pyrimethanil. However, pyrimethanil severely suppressed B. marinus B-9987, whereas the lipopeptides promoted swarming motility and biocontrol efficacy of Bacillus biomass. Lipopeptides at 87.51% purity were tested for systemic acute toxicity and confirmed as low-toxicity substances. In conclusion, low-toxicity lipopeptides were potential alternatives to agrochemicals, and they also performed good promotion when combined with homologous biological control microorganism. There were 2 breakthroughs in this research: (1) marine-derived bacterial lipopeptides inhibited grey mold caused by B. cinerea in field experiments; and (2) purified bacterial lipopeptides (sample purity: >87.51%) were determined to be low-toxicity substances by systemic acute toxicity tests, satisfying the strict requirement of pesticide registration in China (required purity: >85%). This study provides support for using extracellular Bacillus-derived lipopeptides commercially similar to Bacillus-based biological control agents.