Yung Chung Lo

Dark fermentative hydrogen production from enzymatic hydrolysate of xylan and pretreated rice straw by Clostridium butyricum CGS5

Xylan and rice straw were used to produce H(2) via a two-stage approach combining feedstock pretreatment/hydrolysis and dark H(2) fermentation. Acinetobacter junii F6-02 was used to produce cellulolytic enzymes (mainly xylanase) to hydrolyze xylan and pretreated rice straw. The hydrolysates were converted to H(2) by Clostridium butyricum CGS5 via dark fermentation. Investigation of kinetics of xylanase on xylan and NaOH-pretreated rice straw shows nu(max) values of 8.6 and 3.6g/L/h, and K(m) values of 10.6 and 26.9 g/L, respectively. A maximum hydrogen production rate of 62.5 and 26.8 ml/h/L was obtained from hydrolysate of xylan and pretreated rice straw, respectively, while the hydrogen yield was 0.70 and 0.76 mol H(2)/mol xylose, respectively. Simultaneous saccharification and BioH(2) fermentation from xylan was also conducted but giving a lower hydrogen production rate (35.3 ml/h/L) than that of the two-stage process.

Cellulosic hydrogen production with a sequencing bacterial hydrolysis and dark fermentation strategy

In this study, cellulose hydrolysis activity of two mixed bacterial consortia (NS and QS) was investigated. Combination of NS culture and BHM medium exhibited better hydrolytic activity under the optimal condition of 35 degrees C, initial pH 7.0, and 100rpm agitation. The NS culture could hydrolyze carboxymethyl cellulose (CMC), rice husk, bagasse and filter paper, among which CMC gave the best hydrolysis performance. The CMC hydrolysis efficiency increased with increasing CMC concentration from 5 to 50g/l. With a CMC concentration of 10g/l, the total reducing sugar (RS) production and the RS producing rate reached 5531.0mg/l and 92.9mg/l/h, respectively. Furthermore, seven H2-producing bacterial isolates (mainly Clostridium species) were used to convert the cellulose hydrolysate into H2 energy. With an initial RS concentration of 0.8g/l, the H2 production and yield was approximately 23.8ml/l and 1.21mmol H2/g RS (0.097mmol H2/g cellulose), respectively.

Simultaneous production of 2,3-butanediol, ethanol and hydrogen with a Klebsiella sp strain isolated from sewage sludge

A Klebsiella sp. HE1 strain isolated from hydrogen-producing sewage sludge was examined for its ability to produce H2 and other valuable soluble metabolites (e.g., ethanol and 2,3-butanediol) from sucrose-based medium. The effect of pH and carbon substrate concentration on the production of soluble and gaseous products was investigated. The major soluble metabolite produced from Klebsiella sp. HE1 was 2,3-butanediol, accounting for over 42-58% of soluble microbial products (SMP) and its production efficiency enhanced after increasing the initial culture pH to 7.3 (without pH control). The HE1 strain also produced ethanol (contributing to 29-42% of total SMP) and a small amount of lactic acid and acetic acid. The gaseous products consisted of H2 (25-36%) and CO2 (64-75%). The optimal cumulative hydrogen production (2.7 l) and hydrogen yield (0.92mol H2 mol sucrose(-1)) were obtained at an initial sucrose concentration of 30g CODl(-1) (i.e., 26.7gl(-1)), which also led to the highest production rate for H2 (3.26mmol h(-1)l(-1)), ethanol (6.75mmol h(-1)l(-1)) and 2,3-butanediol (7.14mmol h(-1)l(-1)). The highest yield for H2, ethanol and 2,3-butanediol was 0.92, 0.81 and 0.59molmol-sucrose(-1), respectively. As for the overall energy production performance, the highest energy generation rate was 27.7kJ h(-1)l(-1) and the best energy yield was 2.45kJmolsucrose(-1), which was obtained at a sucrose concentration of 30 and 20g CODl(-1), respectively.

Perspectives on cultivation strategies and photobioreactor designs for photo-fermentative hydrogen production

Photosynthetic bacteria have considerable biotechnological potential for biological hydrogen production due to higher substrate conversion efficiency and hydrogen yield. Phototrophic fermentation using photosynthetic bacteria has a major advantage of being able to further convert the byproducts originating from dark fermentation (e.g., volatile fatty acids) to hydrogen. Through the combination of dark and photo-fermentation processes, organic feedstock is fully converted into gaseous product (H2) at the highest possible H2 yield, with significant reduction of chemical oxygen demand (COD). The performance of photo-fermentation is highly dependent on the medium composition, culture conditions, and photobioreactor design. Therefore, this article provides a critical review of the effects of key factors affecting the photo-hydrogen production efficiency of photosynthetic bacteria, and also summarizes the strategies being applied in promoting the performance of photo-fermentation.

Strategies to enhance cell growth and achieve high-level oil production of a Chlorella vulgaris isolate.

The autotrophic growth of an oil‐rich indigenous microalgal isolate, identified as Chlorella vulgaris CC, was promoted by using engineering strategies to obtain the microalgal oil for biodiesel synthesis. Illumination with a light/dark cycle of 14/10 (i.e., 14 h light‐on and 10 h light‐off) resulted in a high overall oil production rate (voil) of 9.78 mg/L/day and a high electricity conversion efficiency (Ec) of 23.7 mg cell/kw h. When using a NaHCO3 concentration of 1,500 mg/L as carbon source, the voil and Ec were maximal at 100 mg/L/day and 128 mg/kw h, respectively. A Monod type model was used to describe the microalgal growth kinetics with an estimated maximum specific growth rate (μmax) of 0.605 day−1 and a half saturation coefficient (Ks) of 124.9 mg/L. An optimal nitrogen source (KNO3) concentration of 625 mg/L could further enhance the microalgal biomass and oil production, leading to a nearly 6.19 fold increase in voil value.

Cultivation of Chlorella vulgaris JSC-6 with swine wastewater for simultaneous nutrient/COD removal and carbohydrate production

Swine wastewater, containing a high concentration of COD and ammonia nitrogen, is suitable for the growth of microalgae, leading to simultaneous COD/nutrients removal from the wastewater. In this study, an isolated carbohydrate-rich microalga Chlorella vulgaris JSC-6 was adopted to perform swine wastewater treatment. Nearly 60-70% COD removal and 40-90% NH3-N removal was achieved in the mixotrophic and heterotrophic culture, depending on the dilution ratio of the wastewater, while the highest removal percentage was obtained with 20-fold diluted wastewater. Mixotrophic cultivation by using fivefold diluted wastewater resulted in the highest biomass concentration of 3.96 g/L. The carbohydrate content of the microalga grown on the wastewater can reach up to 58% (per dry weight). The results indicated that the microalgae-based wastewater treatment can efficiently reduce the nutrients and COD level, and the resulting microalgal biomass had high carbohydrate content, thereby having potential applications for the fermentative production of biofuels or chemicals.

Dark hydrogen fermentation from hydrolyzed starch treated with recombinant amylase originating from Caldimonas taiwanensis On1.

Starch is one of the most abundant resources on earth and is suited to serve as a cost‐effective feedstock for biological hydrogen production. However, producing hydrogen from direct fermentation of starch is usually inefficient, as the starch hydrolysis is often the rate‐limiting step. Therefore, in the present work, enzymatic starch hydrolysis was conducted to enhance the feasibility of using starch feedstock for H2 production. The amylase (with a molecular weight of ca. 112 kDa) used for starch hydrolysis was produced from a recombinant E. coli harboring an amylase gene originating from Caldimonas taiwanensis On1. Using statistical experimental design, the optimal pH and temperature for starch hydrolysis with the recombinant amylase was pH 6.86 and 52.4 °C, respectively, at an initial starch concentration of 7 g/L. The hydrolyzed products contained mainly glucose, maltotriose, and maltotetrose, while a tiny amount of maltose was also detected. The enzymatically hydrolyzed products of soluble starch and cassava starch were used as the substrate for dark hydrogen fermentation using Clostridium butyricum CGS2 and Clostridium pasteurianum CH4. The highest H2 production rate (vH2) and yield (YH2) of C. butyricum CGS2 was 124.0 mL/h/L and 6.32 mmol H2/g COD, respectively, both obtained with the hydrolysate of cassava starch. The best H2 production rate (63.0 mL/h/L) of C. pasteurianum CH4 occurred when using hydrolyzed cassava starch as the substrate, whereas the highest yield (9.95 mmol H2/g COD) was obtained with the hydrolyzed soluble starch.

Improving protein production of indigenous microalga Chlorella vulgaris FSP-E by photobioreactor design and cultivation strategies

Fish meal is currently the major protein source for commercial aquaculture feed. Due to its unstable supply and increasing price, fish meal is becoming more expensive and its availability is expected to face significant challenges in the near future. Therefore, feasible alternatives to fish meal are urgently required. Microalgae have been recognized as the most promising candidates to replace fish meal because the protein composition of microalgae is similar to fish meal and the supply of microalgae-based proteins is sustainable. In this study, an indigenous microalga (Chlorella vulgaris FSP-E) with high protein content was selected, and its feasibility as an aquaculture protein source was explored. An innovative photobioreactor (PBR) utilizing cold cathode fluorescent lamps as an internal light source was designed to cultivate the FSP-E strain for protein production. This PBR could achieve a maximum biomass and protein productivity of 699 and 365 mg/L/day, respectively, under an optimum urea and iron concentration of 12.4 mM and 90 μM, respectively. In addition, amino acid analysis of the microalgal protein showed that up to 70% of the proteins in this microalgal strain consist of indispensable amino acids. Thus, C. vulgaris FSP-E appears to be a viable alternative protein source for the aquaculture industry.

Exploring the inhibitory characteristics of acid hydrolysates upon butanol fermentation: A toxicological assessment.

This study aimed to quantitatively evaluate the inhibitor tolerance of butanol-producing bacterium Clostridium acetobutylicum. The inhibitory effect of the inhibitors generated by acid pretreatment of biomass feedstock on butanol fermentation decreased in the order of formic acid>oxalic acid>furfural>5-HMF>Na2SO4. C. acetobutylicum has a small tolerance range for furfural (1.06-2.6g/L) and 5-HMF (1.99-2.3g/L). However, the inhibitory effect of Na2SO4 appears to have a wide range, with a chronic toxicity for C. acetobutylicum. All the results could explain, in quantitative manner, the instability of butanol fermentation with C. acetobutylicum from various acid-pretreated feedstocks caused by the fermentation inhibitors.

Recovery of gold from industrial wastewater by extracellular proteins obtained from a thermophilic bacterium Tepidimonas fonticaldi AT-A2

Biosorption has emerged as a promising alternative approach for treating wastewater with dilute metal contents in a green and cost effective way. In this study, extracellular proteins of an isolated thermophilic bacterium (Tepidimonas fonticaldi AT-A2) were used as biosorbent to recover precious metal (i.e., Au) from wastewater. The Au (III) adsorption capacity on the T. fonticaldi AT-A2 proteins was the highest when the pH was set at about 4.0-5.0. The adsorption capacity increased with increasing temperature from 15 to 70°C. Adsorption isotherm studies show that both Langmuir and Freundrich models could describe the adsorption equilibrium. The maximum adsorption capacity of Au (III) at 50°C and pH 5 could reach 9.7mg Au/mg protein. The protein-based biosorbent was also used for the recovery of Au from a wastewater containing 15mg/L of Au, achieving a high adsorption capacity of 1.45mg Au/mg protein and a removal efficiency of 71%.