Effect of Solid-State Fermented Brown Rice Extracts on 3T3-L1 Adipocyte Differentiation.

Aspergillus oryzae KCCM 11372 was used to enhance the production of ß-glucan using humidity control strategies. Under conditions of 60% humidity, solid-state fermentation (SSF) increased the yields of enzymes (amylase and protease), fungal biomass (ergosterol), and ß-glucan. The maximum concentrations obtained were 14800.58 U/g at 72 h, 1068.14 U/g at 120 h, 1.42 mg/g at 72 h, and 12.0% (w/w) at 72 h, respectively. Moreover, the ß-glucan containing fermented brown rice (ß-glucan-FBR) extracts at concentrations of 25-300 µg/ml was considered noncytotoxic to 3T3-L1 preadipocytes. We then studied the inhibitory effects of the extracts on fat droplet formation in 3T3-L1 cells. As a result, 300 µg/ml of ß-glucan-FBR extracts showed a high inhibition of 38.88% in lipid accumulation. Further, these extracts inhibited adipogenesis in the 3T3-L1 adipocytes by decreasing the expression of C/EBPα, PPARγ, aP2, and GLUT4 genes.

Synbiotic Fermentation of Undaria pinnatifida and Lactobacillus brevis to Produce Prebiotics and Probiotics.

It has been optimized thermal acid hydrolytic pretreatment and enzymatic saccharification (Es) in flask culture of Undaria pinnatifida seaweed, which is a prebiotic. The optimal hydrolytic conditions were a slurry content of 8% (w/v), 180 mM H2SO4, and 121°C for 30 min. Es using Celluclast 1.5 L at 8 U/mL produced 2.7 g/L glucose with an efficiency of 96.2%. The concentration of fucose (a prebiotic) was 0.48 g/L after pretreatment and saccharification. The fucose concentration decreased slightly during fermentation. Monosodium glutamate (MSG) (3%, w/v) and pyridoxal 5'-phosphate (PLP) (30 µM) were added to enhance gamma-aminobutyric acid (GABA) production. To further improve the consumption of mixed monosaccharides, adaptation of Lactobacillus brevis KCL010 to high concentrations of mannitol improved the synbiotic fermentation efficiency of U. pinnatifida hydrolysates.

Comparison of Liquid and Solid-State Fermentation Processes for the Production of Enzymes and Beta-Glucan from Hulled Barley.

Solid-state fermentation using hulled barley was carried out to produce enzymes and ß-glucan. The one-factor-at-a-time experiments were carried out to determine the optimal composition of the basal medium. The modified synthetic medium composition in liquid-state fermentation was determined to be 70 g/l hulled barley, 0 g/l rice bran, 5 g/l soytone, and 6 g/l ascorbic acid. Optimal pretreatment conditions of hulled barley by solid-state fermentation were evaluated in terms of maximum production of fungal biomass, amylase, protease, and ß-glucan, which were 1.26 mg/g, 31310.34 U/g, 2614.95 U/g, and 14.6% (w/w), respectively, at 60 min of pretreatment condition. Thus, the solid-state fermentation process was found to enhance the overall fermentation yields of hulled barley to produce high amounts of enzymes and ß-glucan.

Coproduction of Enzymes and Beta-Glucan by Aspergillus oryzae Using Solid-State Fermentation of Brown Rice.

The effect of medium composition on enzyme and ß-glucan production by Aspergillus oryzae KCCM 12698 was investigated. Brown rice, rice bran, nitrogen, and ascorbic acid are key components of the synthetic medium used in liquid-state fermentation. To determine the optimal concentrations of these components for enzyme and ß-glucan production, we conducted one factor at a time experiments, which showed that the optimal concentrations were 30 g/l brown rice, 30 g/l rice bran, 10 g/l soytone, and 3 g/l ascorbic acid. Pretreatment of brown rice for 60 min prior to inoculation enhanced fungal biomass, while increasing the production of enzymes and ß-glucan using solidstate fermentation. Maximum fungal biomass of 0.76 mg/g, amylase (26,551.03 U/g), protease (1,340.50 U/g), and ß-glucan at 9.34% (w/w) were obtained during fermentation. Therefore, solidstate fermentation of brown rice is a process that could enhance yield and overall production of enzymes and ß-glucan for use in various applications.

Evaluation of gamma-aminobutyric acid (GABA) production by Lactobacillus plantarum using two-step fermentation.

Lactic acid bacteria (Lactobacillus plantarum KCTC 3103) were fermented to produce gamma-aminobutyric acid (GABA). The conditions of the modified synthetic medium were optimized as 5 g/L glucose, 10 g/L yeast extract, 100 g/L rice bran extract, and 1.0 g/L ascorbic acid for GABA production. Single-step fermentation of cell growth and GABA production with a modified synthetic medium was higher than those with an MRS medium. Two-step fermentation was evaluated by separating the cell growth and GABA production under a modified synthetic medium. The cell concentration of 1.65 g dcw/L produced by the modified synthetic medium was higher than that of 1.0 g dcw/L produced by the MRS medium at 36 h from the first step of two-step fermentation. The highest GABA production of L. plantarum KCTC 3103 was 0.67 g/L with monosodium glutamate addition at 60 h in the second step of fermentation. Two-step fermentation with the modified synthetic medium is suitable for GABA production because of its high GABA productivity and favorable cell growth.

Evaluation of 2,3-Butanediol Production from Red Seaweed Gelidium amansii Hydrolysates Using Engineered Saccharomyces cerevisiae.

Hyper-thermal (HT) acid hydrolysis of red seaweed Gelidium amansii was performed using 12% (w/v) slurry and an acid mix concentration of 180 mM at 150°C for 10 min. Enzymatic saccharification when using a combination of Celluclast 1.5 L and CTec2 at a dose of 16 U/ml led to the production of 12.0 g/l of reducing sugar with an efficiency of enzymatic saccharification of 13.2%. After the enzymatic saccharification, 2,3-butanediol (2,3-BD) fermentation was carried out using an engineered S. cerevisiae strain. The use of HT acid-hydrolyzed medium with 1.9 g/l of 5-hydroxymethylfurfural showed a reduction in the lag time from 48 to 24 h. The 2,3-BD concentration and yield coefficient at 72 h were 14.8 g/l and 0.30, respectively. Therefore, HT acid hydrolysis and the use of the engineered S. cerevisiae strain can enhance the overall 2,3-BD yields from G. amansii seaweed.

Correction to: Butanol and butyric acid production from Saccharina japonica by Clostridium acetobutylicum and Clostridium tyrobutyricum with adaptive evolution.

Unfortunately, the author name was wrongly published as Pailin Sukwang.instead of Pailin Sukwong.

Butanol and butyric acid production from Saccharina japonica by Clostridium acetobutylicum and Clostridium tyrobutyricum with adaptive evolution.

Optimal conditions of hyper thermal (HT) acid hydrolysis of the Saccharina japonica was determined to a seaweed slurry content of 12% (w/v) and 144 mM H2SO4 at 160 °C for 10 min. Enzymatic saccharification was carried out at 50 °C and 150 rpm for 48 h using the three enzymes at concentrations of 16 U/mL. Celluclast 1.5 L showed the lowest half-velocity constant (Km) of 0.168 g/L, indicating a higher affinity for S. japonica hydrolysate. Pretreatment yielded a maximum monosaccharide concentration of 36.2 g/L and 45.7% conversion from total fermentable monosaccharides of 79.2 g/L with 120 g dry weight/L S. japonica slurry. High cell densities of Clostridium acetobutylicum and Clostridium tyrobutyricum were obtained using the retarding agents KH2PO4 (50 mM) and NaHCO3 (200 mM). Adaptive evolution facilitated the efficient use of mixed monosaccharides. Therefore, adaptive evolution and retarding agents can enhance the overall butanol and butyric acid yields from S. japonica.

Application of the Severity Factor and HMF Removal of Red Macroalgae Gracilaria verrucosa to Production of Bioethanol by Pichia stipitis and Kluyveromyces marxianus with Adaptive Evolution.

Gracilaria verrucosa, red seaweed, is a promising biomass for bioethanol production due to its high carbohydrate content. The optimal hyper thermal (HT) acid hydrolysis conditions are 12% (w/v) G. verrucosa with 0.2 M H2SO4 at 130 °C for 15 min, with a severity factor of 1.66. This HT acid hydrolysis produces 50.7 g/L monosaccharides. The maximum monosaccharide concentration of 58.0 g/L was achieved with 96.6% of the theoretical monosaccharide production from 120 g dry weight/L G. verrucosa slurry after HT acid hydrolysis and enzymatic saccharification. Fermentation was carried out by removing an inhibitory compound and via yeast adaptation to galactose. Both Pichia stipitis and Kluyveromyces marxianus adapted to galactose were excellent producers, with the ethanol yield (YEtOH) of 0.50 and 29.0 g/L ethanol production. However, the bioethanol productivity with Pichia stipitis adapted to galactose is higher than that with Kluyveromyces marxianus adapted to galactose, being 0.81 and 0.35 g/L/h, respectively. The results from this study can be applied to industrial scale bioethanol production from seaweed.

Acetone, butanol, and ethanol production from the green seaweed Enteromorpha intestinalis via the separate hydrolysis and fermentation.

Acetone, butanol, and ethanol (ABE) were produced following the separate hydrolysis and fermentation (SHF) method using polysaccharides from the green macroalgae Enteromorpha intestinalis as biomass. We focused on the optimization of enzymatic saccharification as pretreatments for the fermentation of E. intestinalis. Pretreatment was carried out with 10% (w/v) seaweed slurry and 270-mM H2SO4 at 121 °C for 60 min. Monosaccharides (mainly glucose) were obtained from enzymatic hydrolysis with a 16-U/mL mixture of Celluclast 1.5 L and Viscozyme L at 45 °C for 36 h. ABE fermentation with 10% (w/v) E. intestinalis hydrolysate was performed using the anaerobic bacteria Clostridium acetobutylicum with either uncontrolled pH, pH controlled at 6.0, or pH controlled initially at 6.0 and then 4.5 after 4 days, which produced ABE contents of 5.6 g/L with an ABE yield (YABE) of 0.24 g/g, 4.8 g/L with an YABE of 0.2 g/g, and 8.5 g/L with an YABE of 0.36 g/g, respectively.

Improved fermentation performance to produce bioethanol from Gelidium amansii using Pichia stipitis adapted to galactose.

This study employed a statistical method to obtain optimal hyper thermal acid hydrolysis conditions using Gelidium amansii (red seaweed) as a source of biomass. The optimal hyper thermal acid hydrolysis using G. amansii as biomass was determined as 12% (w/v) slurry content, 358.3 mM H2SO4, and temperature of 142.6 °C for 11 min. After hyper thermal acid hydrolysis, enzymatic saccharification was carried out. The total monosaccharide concentration was 45.1 g/L, 72.2% of the theoretical value of the total fermentable monosaccharides of 62.4 g/L based on 120 g dry weight/L in the G. amansii slurry. To increase ethanol production, 3.8 g/L 5-hydroxymethylfurfural (HMF) in the hydrolysate was removed by treatment with 3.5% (w/v) activated carbon for 2 min and fermented with Pichia stipitis adapted to high galactose concentrations via separate hydrolysis and fermentation. With complete HMF removal and the use of P. stipitis adapted to high galactose concentrations, 22 g/L ethanol was produced (yield 0.50). Fermentation with total HMF removal and yeast adapted to high galactose concentrations increased the fermentation performance and decreased the fermentation time from 96 to 36 h compared to traditional fermentation.

Acetone-Butanol-Ethanol Production from Waste Seaweed Collected from Gwangalli Beach, Busan, Korea, Based on pH-Controlled and Sequential Fermentation Using Two Strains.

The optimal conditions for acetone-butanol-ethanol (ABE) production were evaluated using waste seaweed from Gwangalli Beach, Busan, Korea. The waste seaweed had a fiber and carbohydrate, content of 48.34%; these are the main resources for ABE production. The optimal conditions for obtaining monosaccharides based on hyper thermal (HT) acid hydrolysis of waste seaweed were slurry contents of 8%, sulfuric acid concentration of 138 mM, and treatment time of 10 min. Enzymatic saccharification was performed using 16 unit/mL Viscozyme L, which showed the highest affinity (Km = 1.81 g/L). After pretreatment, 34.0 g/L monosaccharides were obtained. ABE fermentation was performed with single and sequential fermentation of Clostridium acetobutylicum and Clostridium tyrobutyricum; this was controlled for pH. A maximum ABE concentration of 12.5 g/L with YABE 0.37 was achieved using sequential fermentation with C. tyrobutyricum and C. acetobutylicum. Efficient ABE production from waste seaweed performed using pH-controlled culture broth and sequential cell culture.

Effects of wavelength mixing ratio and photoperiod on microalgal biomass and lipid production in a two-phase culture system using LED illumination.

Blue and red light-emitting diodes (LEDs) were used to study the effects of wavelength mixing ratios, photoperiod regimes, and green wavelength stress on Nannochloropsis salina, Isochrysis galbana, and Phaeodactylum tricornutum cell biomass and lipid production. The maximum specific growth rates of I. galbana and P. tricornutum were obtained under a 50:50 mixing ratio of blue and red wavelength LEDs; that of N. salina was obtained under red LED. Maximum cell biomass for N. salina and P. tricornutum was 0.75 and 1.07 g dcw/L, respectively, obtained under a 24:0 h light/dark cycle. However, the maximum I. galbana biomass was 0.89 g dcw/L under an 18:6 h light/dark cycle. The maximum lipid contents for N. salina, I. galbana, and P. tricornutum were 49.4, 63.3 and 62.0% (w/w), respectively, after exposure to green LED. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were obtained 1% in P. tricornutum and 2% in I. galbana.

Bioethanol Production from Soybean Residue via Separate Hydrolysis and Fermentation.

Bioethanol was produced using polysaccharide from soybean residue as biomass by separate hydrolysis and fermentation (SHF). This study focused on pretreatment, enzyme saccharification, and fermentation. Pretreatment to obtain monosaccharide was carried out with 20% (w/v) soybean residue slurry and 270 mmol/L H2SO4 at 121 °C for 60 min. More monosaccharide was obtained from enzymatic hydrolysis with a 16 U/mL mixture of commercial enzymes C-Tec 2 and Viscozyme L at 45 °C for 48 h. Ethanol fermentation with 20% (w/v) soybean residue hydrolysate was performed using wild-type and Saccharomyces cerevisiae KCCM 1129 adapted to high concentrations of galactose, using a flask and 5-L fermenter. When the wild type of S. cerevisiae was used, an ethanol production of 20.8 g/L with an ethanol yield of 0.31 g/g consumed glucose was obtained. Ethanol productions of 33.9 and 31.6 g/L with ethanol yield of 0.49 g/g consumed glucose and 0.47 g/g consumed glucose were obtained in a flask and a 5-L fermenter, respectively, using S. cerevisiae adapted to a high concentration of galactose. Therefore, adapted S. cerevisiae to galactose could enhance the overall ethanol fermentation yields compared to the wild-type one.

Effects of light-emitting diode (LED) with a mixture of wavelengths on the growth and lipid content of microalgae.

Integrations of two-phase culture for cell growth and lipid accumulation using mixed LED and green LED wavelengths were evaluated with the microalgae, Phaeodactylum tricornutum, Isochrysis galbana, Nannochloropsis salina, and Nannochloropsis oceanica. Among the single and mixed LED wavelengths, mixed LED produced higher biomass of the four microalgae, reaching 1.03 g DCW/L I. galbana, followed by 0.95 g DCW/L P. tricornutum, 0.85 g DCW/L N. salina, and 0.62 g DCW/L N. oceanica than single LED or fluorescent lights at day 10. Binary combination of blue and red LEDs could produce the high biomass and photosynthetic pigments in the four microalgae. The highest lipid accumulation during second phase with the exposure to green LED wavelengths was 56.0% for P. tricornutum, 55.2% for I. galbana, 53.0% for N. salina, and 51.0% for N. oceanica. The major fatty acid in the four microalgae was palmitic acid (C16:0) accounting for 38.3-47.3% (w/w) of the total fatty acid content.

Bioethanol Production Using Waste Seaweed Obtained from Gwangalli Beach, Busan, Korea by Co-culture of Yeasts with Adaptive Evolution.

Conditions for ethanol production were evaluated using waste seaweed obtained from Gwangalli beach, Busan, Korea, after strong winds on January 15, 2015. Eleven types of seaweed were identified, and the proportions of red, brown, and green seaweed wastes were 26, 46, and 28%, respectively. Optimal pretreatment conditions were determined as 8% slurry content, 286 mM H2SO4 for 90 min at 121 °C. Enzymatic saccharification with 16 units/mL Celluclast 1.5L and Viscozyme L mixture at 45 °C for 48 h was carried out as optimal condition. A maximum monosaccharide concentration of 30.2 g/L was obtained and used to produce ethanol. Fermentation was performed with single or mixed yeasts of non-adapted and adapted Saccharomyces cerevisiae KCTC 1126 and Pichia angophorae KCTC 17574 to galactose and mannitol, respectively. The maximum ethanol concentration and yield of 13.5 g/L and YEtOH of 0.45 were obtained using co-culture of adapted S. cerevisiae and P. angophorae.

Bioethanol production from Gracilaria verrucosa using Saccharomyces cerevisiae adapted to NaCl or galactose.

This study examined the pretreatment, enzymatic saccharification, and fermentation of the red macroalgae Gracilaria verrucosa using adapted saccharomyces cerevisiae to galactose or NaCl for the increase of bioethanol yield. Pretreatment with thermal acid hydrolysis to obtain galactose was carried out with 11.7% (w/v) seaweed slurry and 373 mM H2SO4 at 121 °C for 59 min. Glucose was obtained from enzymatic hydrolysis. Enzymatic saccharification was performed with a mixture of 16 U/mL Celluclast 1.5L and Viscozyme L at 45 °C for 48 h. Ethanol fermentation in 11.7% (w/v) seaweed hydrolysate was carried out using Saccharomyces cerevisiae KCTC 1126 adapted or non-adapted to high concentrations of galactose or NaCl. When non-adapted S. cerevisiae KCTC 1126 was used, the ethanol productivity was 0.09 g/(Lh) with an ethanol yield of 0.25. Ethanol productivity of 0.16 and 0.19 g/(Lh) with ethanol yields of 0.43 and 0.48 was obtained using S. cerevisiae KCTC 1126 adapted to high concentrations of galactose and NaCl, respectively. Adaptation of S. cerevisiae KCTC 1126 to galactose or NaCl increased the ethanol yield via adaptive evolution of the yeast.

Efficient utilization of Eucheuma denticulatum hydrolysates using an activated carbon adsorption process for ethanol production in a 5-L fermentor.

A total monosaccharide concentration of 37.8 g/L and 85.9% conversion from total fermentable monosaccharides of 44.0 g/L from 110 g dw/L Eucheuma denticulatum slurry were obtained by thermal acid hydrolysis and enzymatic saccharification. Subsequent adsorption treatment to remove 5-hydroxymethylfurfural (5-HMF) using 5% activated carbon and an adsorption time of 10 min were used to prevent a prolonged lag phase, reduced cell growth, and low ethanol production. The equilibrium adsorption capacity (q e) of HMF (58.183 mg/g) showed high affinity to activated carbon comparing to those of galactose (2.466 mg/g) and glucose (2.474 mg/g). The efficiency of cell growth and ethanol production with activated carbon treatment was higher than that without activated carbon treatment. Fermentation using S. stipitis KCTC7228 produced a cell concentration of 3.58 g dw/L with Y X/S of 0.107, and an ethanol concentration of 15.8 g/L with Y P/S of 0.48 in 96 h.

Hyper-thermal acid hydrolysis and adsorption treatment of red seaweed, Gelidium amansii for butyric acid production with pH control.

Optimal hyper-thermal (HT) acid hydrolysis conditions for Gelidium amansii were determined to be 12% (w/v) seaweed slurry content and 144 mM H2SO4 at 150 °C for 10 min. HT acid hydrolysis-treated G. amansii hydrolysates produced low concentrations of inhibitory compounds and adsorption treatment using 3% activated carbon. An adsorption time of 5 min was subsequently used to remove the inhibitory 5-hydroxymethylfurfural from the medium. A final maximum monosaccharide concentration of 44.6 g/L and 79.1% conversion from 56.4 g/L total fermentable monosaccharides with 120 g dw/L G. amansii slurry was obtained from HT acid hydrolysis, enzymatic saccharification, and adsorption treatment. This study demonstrates the potential for butyric acid production from G. amansii hydrolysates under non-pH-controlled as well as pH-controlled fermentation using Clostridium acetobutylicum KCTC 1790. The activated carbon treatment and pH-controlled fermentation showed synergistic effects and produced butyric acid at a concentration of 11.2 g/L after 9 days of fermentation.

Enhanced biomass production and lipid accumulation of Picochlorum atomus using light-emitting diodes (LEDs).

The effects of light-emitting diode (LED) wavelength, light intensity, nitrate concentration, and time of exposure to different LED wavelength stresses in a two-phase culture on lipid production were evaluated in the microalga, Picochlorum atomus. The biomass produced by red LED light was higher than that produced by purple, blue, green, or yellow LED and fluorescent lights from first phase of two-phase culture. The highest lipid production of P. atomus was 50.3% (w/w) with green LED light at 2days of second phase as light stress. Fatty acid analysis of the microalgae showed that palmitic acid (C16:0) and linolenic acid (C18:3) accounted for 84-88% (w/w) of total fatty acids from P. atomus. The two-phase culture of P. atomus is suitable for biofuel production due to higher lipid productivity and favorable fatty acid composition.