Shih-Hsin Ho

Effect of light intensity and nitrogen starvation on CO2 fixation and lipid/carbohydrate production of an indigenous microalga Scenedesmus obliquus CNW-N

Engineering strategies were applied to improve the CO(2) fixation rate and carbohydrate/lipid production of a Scenedesmus obliquus CNW-N isolate. The light intensity that promotes cell growth, carbohydrate/lipid productivity, and CO(2) fixation efficiency was identified. Nitrogen starvation was also employed to trigger the accumulation of lipid and carbohydrate. The highest productivity of biomass, lipid, and carbohydrate was 840.57 mg L(-1)d(-1), 140.35 mg L(-1)d(-1). The highest lipid and carbohydrate content was 22.4% (5-day N-starvation) and 46.65% (1-day N-starvation), respectively. The optimal CO(2) consumption rate was 1420.6 mg L(-1)d(-1). This performance is better than that reported in most other studies. Under nitrogen starvation, the microalgal lipid was mainly composed of C16/C18 fatty acid (around 90%), which is suitable for biodiesel synthesis. The carbohydrate present in the biomass was mainly glucose, accounting for 77-80% of total carbohydrates. This carbohydrate composition is also suitable for fermentative biofuels production (e.g., bioethanol and biobutanol).

Bioethanol production using carbohydrate-rich microalgae biomass as feedstock.

This study aimed to evaluate the potential of using a carbohydrate-rich microalga Chlorella vulgaris FSP-E as feedstock for bioethanol production via various hydrolysis strategies and fermentation processes. Enzymatic hydrolysis of C. vulgaris FSP-E biomass (containing 51% carbohydrate per dry weight) gave a glucose yield of 90.4% (or 0.461 g (g biomass)(-1)). The SHF and SSF processes converted the enzymatic microalgae hydrolysate into ethanol with a 79.9% and 92.3% theoretical yield, respectively. Dilute acidic hydrolysis with 1% sulfuric acid was also very effective in saccharifying C. vulgaris FSP-E biomass, achieving a glucose yield of nearly 93.6% from the microalgal carbohydrates at a starting biomass concentration of 50 g L(-1). Using the acidic hydrolysate of C. vulgaris FSP-E biomass as feedstock, the SHF process produced ethanol at a concentration of 11.7 g L(-1) and an 87.6% theoretical yield. These findings indicate the feasibility of using carbohydrate-producing microalgae as feedstock for fermentative bioethanol production.

Microalgae-based biorefinery--from biofuels to natural products.

The potential for biodiesel production from microalgal lipids and for CO2 mitigation due to photoautotrophic growth of microalgae have recently been recognized. Microalgae biomass also has other valuable components, including carbohydrates, long chain fatty acids, pigments and proteins. The microalgae-based carbohydrates consist mainly of cellulose and starch without lignin; thus they can be ready carbon source for the fermentation industry. Some microalgae can produce long chain fatty acids (such as DHA and EPA) as valuable health food supplements. In addition, microalgal pigments and proteins have considerable potential for many medical applications. This review article presents comprehensive information on the current state of these commercial applications, as well as the utilization and characteristics of the microalgal components, in addition to the key factors and challenges that should be addressed during the production of these materials, and thus provides a useful report that can aid the development of an efficient microalgae-based biorefinery process.

Characterization and optimization of carbohydrate production from an indigenous microalga Chlorella vulgaris FSP-E

In this study, three indigenous microalgae isolates were examined for their ability to produce carbohydrates. Among them, Chlorella vulgaris FSP-E displayed relatively high cell growth rate and carbohydrate content. The carbohydrate productivity of C. vulgaris FSP-E was further improved by using engineering strategies. The results show that using an appropriate light intensity and inoculum size could effectively promote cell growth and carbohydrate productivity. Nitrogen starvation triggered the accumulation of carbohydrates in the microalga, achieving a carbohydrate content of 51.3% after 4-day starvation. Under the optimal conditions, the highest biomass and carbohydrate productivity were 1.437 and 0.631 g L(-1) d(-1), respectively. This performance is better than that reported in most related studies. Since glucose accounted for nearly 93% of the carbohydrates accumulated in C. vulgaris FSP-E, the microalga is an excellent feedstock for bioethanol fermentation.

Current progress and future prospect of microalgal biomass harvest using various flocculation technologies

Microalgae have been extensively studied for the production of various valuable products. Application of microalgae for the production of renewable energy has also received increasing attention in recent years. However, high cost of microalgal biomass harvesting is one of the bottlenecks for commercialization of microalgae-based industrial processes. Considering harvesting efficiency, operation economics and technological feasibility, flocculation is a superior method to harvest microalgae from mass culture. In this article, the latest progress of various microalgal cell harvesting methods via flocculation is reviewed with the emphasis on the current progress and prospect in environmentally friendly bio-based flocculation. Harvesting microalgae through bio-based flocculation is a promising component of the low-cost microalgal biomass production technology.

Engineering strategies for improving the CO2 fixation and carbohydrate productivity of Scenedesmus obliquus CNW-N used for bioethanol fermentation

Engineering strategies were applied to improve the cell growth, CO2 fixation ability, and carbohydrate productivity of a Scenedesmus obliquus CNW-N isolate. The resulting carbohydrate-rich microalgal biomass was subsequently utilized as feedstock for ethanol fermentation. The microalga was cultivated with 2.5% CO2 in a photobioreactor on different operation modes. Semi-batch operations with 50% replacement of culture medium resulted in the highest CO2 fixation rate (1546.7 mg L(-1) d(-1)), carbohydrate productivity (467.6 mg L(-1) d(-1)), and bioethanol yield (0.202 g/g biomass). This performance is better than most reported values in the literature. The microalgal biomass can accumulate nearly 50% carbohydrates, as glucose accounted for nearly 80% of the total carbohydrate content. This glucose-predominant carbohydrate composition of the microalga is well suited for fermentative bioethanol production. Therefore, using the proposed carbohydrate-rich microalgal biomass both as the carbon sink and as the feedstock provides a feasible alternative to current carbon-reduction and bioethanol-production strategies.

Bioprocess development on microalgae-based CO2 fixation and bioethanol production using Scenedesmus obliquus CNW-N

A two-stage cultivation strategy was applied to achieve greater CO2 fixation and carbohydrate productivity with an indigenous microalga Scenedesmus obliquus CNW-N, which was first cultivated using a nutrient-rich medium to promote cell growth, and was then switched to a nutrient-deficient condition to trigger carbohydrate accumulation. The optimal biomass productivity, carbohydrate productivity, and CO2 fixation rate were 681.4, 352.9, and 1192.5 mg L(-1) d(-1), respectively, with a 51.8% carbohydrate content (based on dry weight). This performance is better than the results in most related studies. The microalgal carbohydrate was mainly composed of glucose, which accounts for nearly 80% of total sugars. Dilute acid hydrolysis with 2% H2SO4 can saccharify the wet microalgal biomass effectively, achieving a glucose yield of 96-98%. Using the acidic hydrolysate of the microalga as feedstock, the separate hydrolysis and fermentation (SHF) process gave an ethanol concentration of 8.55 g L(-1), representing a theoretical yield of nearly 99.8%.

Characterization of flocculating agent from the self-flocculating microalga Scenedesmus obliquus AS-6-1 for efficient biomass harvest

In the present work, the extracellular biopolymers from the self-flocculating microalga Scenedesmus obliquus AS-6-1 were studied. It was revealed that the self-flocculation of the microalgal cells was mediated by cell wall-associated polysaccharides with a molecular weight of 127.9 kDa. Sugar compositions analysis indicated that the monomers consist of glucose, mannose, galatose, rhamnose and fructose with the molar ratio of 8:5:3:2:1. Addition of 0.6 mg/L purified flocculating agent resulted in the fast flocculation of freely suspended cells of S. obliquus and Chlorella vulgaris. The flocculating activity is stable between pH 6 and 8 and at 20-60°C.

Phototrophic cultivation of a thermo-tolerant Desmodesmus sp. for lutein production: Effects of nitrate concentration, light intensity and fed-batch operation

Four indigenous thermo-tolerant Desmodesmus sp. strains were examined for their ability to produce lutein. Among them, Desmodesmus sp. F51 was the best strain for this purpose. The medium composition, nitrate concentration and light intensity were manipulated to improve the phototrophic growth and lutein production of Desmodesmus sp. F51. It was found that a nitrogen-sufficient condition was required for lutein accumulation, while a high light intensity enhanced cell growth but caused a decrease in the lutein content. The best cell growth and lutein production occurred when the light intensity and initial nitrate concentration were 600 μmol/m(2)/s and 8.8 mM, respectively. The fed-batch cultivation strategy was shown to further improve lutein production. The highest lutein productivity (3.56±0.10 mg/L/d) and content (5.05±0.20 mg/g) were obtained when pulse-feeding of 2.2 mM nitrate was employed. This study demonstrated the potential of using Desmodesmus sp. F51 as a lutein producer in practical applications.

Establishment of an efficient genetic transformation system in Scenedesmus obliquus

Scenedesmus obliquus belongs to green microalgae, which is attracting attention as a feedstock for biofuels production and biorefinery as well as in bioremediation of environmental pollutants, making its genetic modifications for more efficient growth and accumulation of aimed metabolites significant. However, the genetic transformation system of S. obliquus is still not well established. In the current work, S. obliquus was transformed via electroporation using a plasmid containing chloramphenicol resistance gene (CAT) as a selectable marker and the green fluorescent protein gene (gfp) as a reporter. Using the optimized transformation conditions, the transformation efficiency was 494±48 positive transgenic clones per 10(6) recipient cells, which is more efficient comparing with those reported in other microalgal transformation studies. Green fluorescence was observed after six months of cultivation, and CAT-specific products were also detected in the transformants by PCR, Southern blot and RT-PCR analysis. This is the first report on establishing such an efficient and stable transformation system for S. obliquus, a prerequisite for both functional genomic studies and strain improvement for other biotechnology applications of this important microalgal species.