Earticle

Some Antarctic algae are known to produce ice binding proteins. IBPs are diverse proteins with the capability of inhibiting ice crystal growth against in subzero environments for their survival. Two microalgae, named AnF0046 and AnF0048, were used in this study. To examine the physiological properties, we carried out a comparative analysis on growth with Chlamydomonas reinhardtii. Growth rate of C. reinhardtii is 1.6 and 1.1 fold higher than those of AnF0046 and AnF0048, respectively. AnF0046 and AnF0048 produced 1.4 and 1.7 fold lower amounts of chlorophylls than that of C. reinhardtii. Extraction of genomic DNA was conducted for confirmation of gene existence related to IBPs. The bands on 1-D gel from either AnF0046 or AnF0048 were comparable with that from C. reinhardtii. To search IBP encoding gene, primers were designed based on IBP isoforms produced by Chlamydomonas sp. CCMP681. The primer designed from isoform1 had the most homologous sequence. A single band at 2.5 kbp was found from overlapping fragments purified. Forward and reverse sequences, each with 950 bp, were obtained after sequencing. The two Antartic microalgae, AnF0046 and AnF0048, found to have IBP coding gene.

A microorganism hydrolyzing carboxymethylcellulose (CMC) was isolated from seawater, identified as Bacillus atrophaeus by analyses of 16S rDNA sequences, and named as B. atrophaeus LBH-18. Rice bran and ammonium peptone were the best combination of carbon and nitrogen sources for the production of the CMCase by B. atrophaeus LBH-18. Based on intuitive analysis and statistical calculations using data from an L25(35) orthogonal design experiment with three factors of rice bran, peptone, and initial pH of medium, rice bran was found to be the most important factor for production of the CMCase by B. atrophaeus LBH-18. Optimal concentrations of rice bran and peptone for production of CMCase were found to be 5.0% and 0.5%, respectively. Optimal initial pH of medium and temperature were 7.0 and 30℃. Effect of salts was also investigated using an orthogonal array method. Potassium phosphate from four salts in the medium was found to be the most important factor for production of the CMCase by B. atrophaeus LBH-18. Productivity of the CMCase by B. atrophaeus LBH-18 was about 120 U/ml under optimized conditions.

Marine bacteria which possess an alginate degrading activity were isolated from seasoned brown seaweeds with fermented anchovies. Small amount of the liquefied substances was diluted with sterilized water and smeared on M9 alginate agar plate. Among the alginate degrading colonies, a plane, rough, opaque like shape colony was chosen. Analysis of 16S ribosomal DNA sequence and constructed phylogenetic tree revealed that the strain was closest to Caulobacter leidyi, Caulobacter endosymbiont, and Sphingomonas sp. We designated this strain as Marine Bacterium MJ3. Strain MJ3 grew best on the minimal agar plate containing 0.8% sodium alginate as a sole carbon source and showed resistant against several antibiotics. When the strain MJ3 was cultured in the minimal media with alginate addition, the alginate lyase activity occurs in the cell lysate not in the culture soup. It is concluded that the MJ3 engulfs macromolecule alginate into the cell and degraded it to oligo-algininate and mono-alginate as an energy source. Acknowledgement: This work was financially supported by the 2008 Busan Techno Park program and the Ministry of Knowledge Economy (MKE) and Korea Industrial Technology Foundation (KOTEF) through the Human Resource Training Project for Strategic Technology.

Trimethylamine (TMA) is a product of decomposition of plants and animals. With its strong fishy odor in low concentrations, TMA is the main agent for the odor often associated with fouling fish, some infections, and bad breath. This study focused on the adsorption of TMA over various nanoporous zeolites for application in the lowtemperature deodorization of fishy odor from raw fish oil. The faujasite (FAU, Si/Al=3), mordenite (MOR, Si/Al=10), and ZSM-5 (MFI, Si/Al=25-335) zeolites were employed. The TMA was adsorbed in a batch type reactor at 50 oC. The FAU zeolite exhibited the high adsorption ability, which, in combination with its wide surface area and pore volume, may have induced the high adsorption ability. Despite having a small surface area of size similar to that of the HMFI zeolite, the H-MOR(10) zeolite exhibited a large TMA adsorption. The H-MOR(10) zeolite has stronger acid sites than those of the other zeolites, and this increased strength was considered to have generated more attractive adsorption with TMA ions, because TMA interacts briskly with cations of acid sites on the zeolites. Approximately 60% of the odor of the raw fish oil was removed by adsorption on the H-FAU(3) zeolite at 50 oC.

Cell growth and lipid production of Chlorella minutissima in fed-batch culture was studied. Two carbon resources, such as CO2 and glucose were used to compare autotrophic and heterotrophic cultivate condition. For fed-batch cultivation, the feeding rate of seawater medium containing glucose was decided to be 0.5 ℓ/day of medium containing glucose. Autotrophic with CO2 showed the maximum cell growth of 9 (g-dry wt./L) and total lipid production of 15%. heterotrophic condition with glucose 10 g/ ℓ showed the maximum cell growth of 15 (g-dry wt./L) and total lipid production of 32%. Also, lipid profiles produced by gas chromatography analysis showed that lipid concentration of C17 and C18 composition is each 16%, 38%. This tells that Chlorella minutissima is a suitable bio-resource to produce the biodiesel. Accordingly, this study showed that the fed-batch cultivation with glucose is most effective for the large scale cultivation for biodiesel production with Chlorella minutissima.

Conventional biopanning protocol has been applied to explore the peptide sequence having affinity to a target compound starting from a phage-displayed peptide library through a batch type adsorption protocol. The basic principal of biopanning protocol is very similar to the general chromatographic separation which is widely used in bioseparation processes. This study was carried out to improve the efficiency of biopanning protocol based on that similarity of both protocols. The biopanning protocol was automated by using a FPLC equipped with monolithic column which virus particles pass through. This new protocol made it possible to develop “chromatographic library panning protocol” which can search for the specific peptide sequence within a short time compared to the conventional biopanning protocol. Pb2+ has been known to be highly toxic to a majority of living entities and thus classified with other heavy metal ions as endocrine disruptors. The iterative passing of phage particles through the Pb2+-combined monolithic column in FPLC was done to find the peptide sequence having high affinity to Pb2+. Then the screened Pb2+- binding phage particles was sequentially loaded to the other monolithic columns combined with other metal ions (Ni2+, Cu2+, Zn2+, Co2+) to remove the peptide sequence having cross-binding affinity to other metal ions.

Biobutanol has many characteristics that make it a better biofuel than bioethanol, now used in the formulation of gasohol. Despite the remarkable advantages of butanol as a fuel, biobutanol production has several limitations, including low values for the yield, productivity, and final product concentration. In this study, we examined the effect of two phase fermentation for the in-situ removal of butanol from fedbatch reactor fermentation broth using Clostridium beijerinckii NCIMB 8052. A non-toxic immiscible solvent, oleyl alcohol, was added to, and removed directly from the fed-batch reactor and extracted the majority of the inhibitory butanol from the aqueous broth. Oleyl alcohol did not inhibit the growth of the fermentative organism. In the batch culture, without solvent extraction, butanol production ceased after 30 h at a concentration of 11.6 g/L. Applying oleyl alcohol as an extraction solvent, about 83% of the total butanol produced was successfully extracted. In addition, improvement of productivity (0.34 g/L·h) and total butaol concentration (24.1 g/L) were achieved, which were 154 % and 206 %, respectively, higher than those obtained in the controlled traditional batch process. In conclusion, the presented two phase fermentation system for in-situ butanol removal by liquid-liquid extraction could be successfully applied for an integrated fermentation/product recovery system.

Solventogenic clostridia lose their ability to produce solvent during repeated sub-culturing of batch culture or during a continuous fermentation. This phenomenon is known as 'degeneration' and degeneration of solventogenic clostridia causes failure of stable butanol production during continuous fermentation. Therefore it is needed to prevent the degeneration of cells and maintain the cells in solventogenic phase. In this study, we investigated the quorum sensing signal in Clostidium acetobutylicum during biobutanol production and the relationships between the phenomenon of degeneration and the quorum sensing signal. By measuring the activity of autoinducer-2 (AI-2), a quorum sensing signal molecule, we found out that solventogenic C. acetobutylicum produces AI-2. However, degenerated C. acetobutylicum did not seem to produce AI-2 during their cell cycles. In addition, the effects of quorum sensing signal molecule analogs to biobutanol production were investigated during batch fermentation of C. acetobutylicum.

Yellow poplar was selected for alcohol-based biofuel production through alkaline hydrolysis, enzymatic saccharification and fermentation. In alkaline hydrolysis, 50.0% of biomass remained as residual, which chemical composition was 82.0% of cellulose, 17.0% xylan and 1.0% of lignin. Enzymatic saccharification led to almost complete conversion of cellulose and xylan in alkaline hydrolysis residue to monosaccharide. After enzymatic saccharification, the composition of hydrolysate was analyzed by HPLC. The sugar composition of enzymatic hydrolysate was 95.1 g/L of glucose and 21.4 g/L of xylose. The enzymatic hydrolysate also contained 0.5 g/L of acetic acid and 0.48 g/L of total phenolics. The furfural and 5-hydroxymethylfurfural (5-HMF) was not detected in this liquor. It means that the fermentation inhibitors such as phenolics and furans are not existed or under control. The hydrolysate from enzymatic saccharification was used for the production of ethanol and butanol by Saccharomyces cerevisiae and Clostridium beijerinckii, respectively. The ethanol production yield for sugar was about 58% in the fermentation of hydrolysate, which shows the same level of the control. In ethanol fermentation, glucose was completely utilized, where xylose was not consumed. The butanol was produced to the level of the control and xylose was consumed a little bit.

Because of the limited stocks of fossil fuels and the production of greenhouse gas on their combustion, alternative sources of energy have been investigated. And following the 'Low Carbon, Green Growth' as Korea's future vision, we need to find a green energy source. One possible source of biological material for fuel production is microalgae. Investigations were carried out on the uptake of lipid by the green alga Chlorella emersonii. In order to get the high lipid content using C. emersonii, a medium optimization experiment was designed and performed. The effect of media composition on the lipid of C. emersonii was examined by fractional factorial design (FFD) and central composite design (CCD). The media components examined include potassium nitrate, potassium phosphate, magnesium sulphate, ferrous sulphate, and trace metal solution. Trace metal solution was used 1 ml of microelements composed of H3BO3 (2.86 g), MnCl24H2O (2.84 g), ZnSO47H2O (0.22 g), Na2MoO42H2O (0.018 g), CuSO45H2O (0.29 g) in 1000 ml distilled water. All experimental sets were cultivated under 5% CO2 balanced with 95% air and under 60 μE/m2s of light intensity. The results indicated that potassium nitrate, potassium phosphate, and trace metal solution were major factors. The optimum concentrations of potassium nitrate, potassium phosphate, and Trace metal solution were found to be 1.63, 1.06 g/L, and 1.35 ml/L respectively, for lipid production. The optimized media was shown a similar cell concentration although fresh cell weights were 19% higher. The lipid production with optimized medium showed a 30% enhanced production than that with original Watanabe medium.

Biofuel production expansion is seen especially in agriculture producing countries such as U.S.A. and Brazil. However, it has drawn international attention due to global price increase of food stock. Therefore, possibility of utilizing unused cellulose from marine algae for biofuel has been studied. Marine algae contains a high percentage of water, possessing a lower production rate compared to land crops. Nonetheless, biofuel production potential is high, comparable to land crops, since productivity per area is high. In order to obtain biofuel, undaria pinnatifida (belonging to macro algae), chlorella vulgaris and chlamydomonas reinhardtii (belonging to micro algae) were used for the pretreatment experiments. The pretreatment was carried out in dilute acid hydrolysis. To find optimal condition, experiments were performed at various temperatures and acid concentrations.

Xylose is a five-carbon sugar abundant in lignocellulosic biomass, so its fermentation is essential for the economic conversion of lignocelluloses to ethanol. Xylose-fermenting yeast strains, V15, V20, P1-3, and P1-6 were isolated from leaf mold in leaf mold using an enrichment technique in a yeast peptone xylose medium and minima medium containing xylose at 30°C. The phenotypic and physiologica characteristics suggested that most of isolated yeast were Candida tropicalis. During shaking flask cultivation, the highest ethanol productivity and yield of V15, V20, P1-3, and P1-6 in YPGX media containing 25 g/l glucose and 25 g/l xylose were 0.46 g/l/h and 0.34 g/g, 0.46 g/l/h and 0.35 g/g, 0.41 g/l/h and 0.28 g/g, and 0.33 g/l/h and 0.19 g/g, respectively. These results suggest that isolated yeasts are potential producer of metabolic engineering to use in ethanol production from lingo-cellulosic biomass. In addition, strain selection through mutagenesis, adaptive evolution using isolated yeasts can also be employed to further improve activity

Cyanobacteria are photosynthetic bacteria whose characteristics are determined by fixing CO2, using photon energy and evolving O2 gas from water splitting. Owing to these ‘environment- friendly’ features, they have potentials to be exploited for clean, renewable fuel production, especially biohydrogen. To improve cyanobacterial strain for efficient hydrogen production, genetic manipulation technique is needed. Thus, in the present work, we constructed a shuttle expression vector for Synechocystis PCC 6803 (Syn6803) as a first footstep. It consists of several parts which came from different sources. The replication origin (oriV), origin of transfer (oriT), and associated genes (repA,B,C) were cloned as a 5.6kbp fragment from RSF-1010, a broad host range plasmid, allowing to replicate autonomously in some cyanobacterial strains including Syn6803. The commercial vectors, pET-28b (Novagen) and pTrcHisC (Invitrogen), provided kanamycin resistance gene, multiple cloning sites (MCS), and other accessory sequences. The nickel-inducible strong promoter was PCRcloned from nickel response operon (nrs) within the genome of Syn6803. The nickel metal for induction is also essential component for the activity of NiFehydrogenase which is the enzyme responsible for hydrogen production. We introduced green fluorescent protein (GFP) gene into the MCS region of the complete vector which was named pRKNrs. We will present functional expression of reporter GFP in Syn6803 strains to confirm successful construction.

As biorefineries replace oil refineries, society and the environment will benefit from a switch from hydrocarbon feedstocks to renewable carbohydrates as a source of energy, materials and chemicals. Lignocellulosic biomass-based ethanol technologies are rapidly evolving and bottlenecks are being identified that need to be overcome to achieve widespread commercialization. The study, in this sense, was carried out ammonia percolation (AP) pretreatment using cassava stem as lignocellulosic biomass, performed enzymatic saccharification and fermented into ethanol by Saccharomyces cerevisiae. As results of the pretreatment, the delignification ratio appeared very efficiently of 45% or more (120°C, 120min) and cellulose which can be converted to ethanol was excellently conserved. Also, the result of the saccharification, glucose is obtained 27.4 g/L by enzymatic digestibility. And ethanol concentration reached to 12.4 g/L in 24 h. This study showed not only total process for bioethanol production from agricultural biomass but also efficient pretreatment of lignocellulosic material.

The operating conditions for a hollow-fiber membrane filter reactor containing methanogens that can utilize carbon dioxide and hydrogen was studied for the reduction of carbon dioxide. Anaerobic sludge was inoculated into each reactor (20% of the working volume). CRS1 reactor was operated at acidic condition (pH 5.0). The pressure of supplied gas mixture was 6.5 ~ 7.5 psi. Methane was produced in the CRS1 reactor (acidic reactor) after 2 hours and the methane percentage in the biogases increased to 80% on day 5. The volume of produced methane was 400 ~ 500 ml/day on average over the test periods. CRS2 reactor was operated at neutral condition (pH 7.0). Supply pressure of mixed gases was 3 ~ 4 psi. Methane was produced in the CRS2 reactor (neutral condition) after 2 hours and the methane percentage in the biogases produced increases to 60% and it was raised to 80% with time. The volume of produced methane was 600 ~ 700 ml/day on average over the test periods. Acetic acid was continuously produced at this reactor and the volume of produced methane of CRS2 reactor was about three times higher than that of CRS1 reactor.

The waste air containing ammonia was treated and a comparative study was performed with the combined system composed of fluidized aerobic reactor and anaerobic reactor. From the experimental results, a high removal efficiency of ammonia of higher than 90% can be achieved at the early state of reactor-run. However, with the increase of operation time this value decreased and maintained at about 80%. In case of the theoretical model prediction, the concentrations of nitrate ion in the aerobic reactor and in the anaerobic reactor varied around 18ppm and 12ppm, respectively, which were about 31% and 8% different from those in case of the actual experiment. Regarding the concentrations of aqueous ammonia, the total aqueous ammonia and ammonium ion, the difference between theoretical prediction and actual experiment are less than 9.25% and 16.93% in aerobic and anaerobic reactors, respectively. Although there exists a slight difference between the experimental data and the predicted value, the results of this study have shown a quite relevant correlation between the theoretical model and the actual experiment. This correlation shows the superiority of the model to predict the essence of phenomenon occurring in the system.

In this study, we utilized a unique strategy for the fed-batch fermentation process using ethanol-tolerant Saccharomyces cerevisiae to achieve a high-level of ethanol production that could be practically applied to industrial ethanol production. During this study, the aeration rate was controlled at 0.0, 0.13, 0.33, and 0.8 vvm to determine the optimal aeration conditions for the production of ethanol. Additionally, nonsterile glucose power was fed during fed-batch ethanol fermentation, and the amount of corn-steep liquor (CSL) in the medium was optimized. When aeration was conducted, the ethanol production and productivity were superior to those when the aeration was not conducted. Specifically, the maximum ethanol production reached approximately 160 g/L ethanol when the reaction was aerated at 0.13 vvm. Ethanol productivity during the early phase of the process increased, then decreased gradually during the later phase. Despite this finding, when aeration was conducted, the overall ethanol yield was not superior to that obtained when aeration was not conducted. In addition, ethanol consumption was observed at aeration rates of 0.3 and 0.8 vvm which resulted in a remarkable decrease in ethanol productivity and yield. However, the use of a much more inexpensive C-source may enable the process to be directed toward improvement of the overall ethanol production and productivity in a process that is aerated at 0.13 vvm. Furthermore, if a repeated fed-batch process in which the withdrawal and fill is conducted prior to 36 hr can be employed, aeration at a rate of 0.33 and/or 0.8 vvm may improve the overall ethanol productivity. Finally, the overall ethanol productivities in this study varied throughout the fed-batch ethanol fermentation, which indicates that the optimal operational strategy for achieving improved ethanol productivity may be set-up according to various aeration rates.

A method for measuring the ethanol concentration in a yeast culture broth was developed using both microtubes and a 96-deepwell microplate. The strategy involved first the solvent extraction of ethanol from the yeast culture broth and measurements of the ethanol concentration using the dichromate oxidation method. Particular focus was made on selecting the extraction solvent as well as determining the measurable range of ethanol concentrations using this solvent extraction-dichromate oxidation method. This method was developed as an assay format in 2.0 ml microtubes and 1.2 ml 96-deepwell microplates, and the ethanol concentration in the batch cultures and fed-batch fermentations was measured. Trin-butyl phosphate (non-alcoholic solvent, density=0.9727, solubility in water=0.028% (w/v)) was used for solvent extraction when measuring the ethanol concentration from the yeast culture broth. The maximum detectable ethanol concentration was 8% (v/v) when 10 g potassium dichromate in 100 ml of 5M sulfuric acid was used. The concentrations determined from the solvent extractiondichromate oxidation methods were remarkably similar to those from gas chromatography, in which samples were prepared from seven experiments, such as four batch cultures and three fed-batch fermentations.

The biodiesel industry has increased because it can be used as an eco-friendly and renewable alternative biodiesel. Because 10 kg of biodiesel yields 1 kg of glycerol, the cost of crude glycerol is rapidly decreased in the world market. There is a great advantage to use crude glycerol for the useful chemical productions from economical and environmental standpoints. In this study, Clostridium pasteurianum DSM 525 was tested for the production of butanol and 1,3-propanediol (1,3-PD) using crude glycerol a biodiesel waste. When crude glycerol without pre-purification was used as a carbon source for suspended batch cultures, the growth and production of butanol/1,3-PD was inhibited compared to those with pure glycerol. To solve this inhibitory effect of crude glycerol, a cell immobilization system was investigated with C. pasteurianum DSM 525. Cells were immobilized on porous polymer media by exchanging cultures to fresh media (containing pure glycerol) every 24 hrs. Interestingly, after successful cell immobilization, the addition of crude glycerol did not inhibit the growth of cells and butanol/1,3-PD production. When 30 g/L of pure glycerol and 10 g/L of crude glycerol was added, 1~2g/L of 1,3-PD and 10.9 g/L of butanol were produced, which were similar to those produced with pure glycerol only (11~12g/L butanol). In conclusion, the production of 1,3-PD and butanol with C. pasteurianum DSM 525 can be achieved with crude glycerol without any pretreatment.

Hexanoic acid, a six-carbon straight-chain fatty acid, is found in trace amounts in various animal fats and oils. Its commercial use is primarily as a precursor for the synthesis of fine chemicals. In this study, hexanoic acid production from galactitol was investigated using microorganisms from activated sludge. For the production of hexanoic acid, sludge was treated at 100℃ for 30 minutes, and then it was used as an inoculum. The major products in liquor phase were acetic acid, butyric acid, and hexanoic acid. The concentration of acetic acid and butyric acid was about 0.5~2.5 g/L. Yield of produced hexanoic acid was about 0.35±0.1 g/ galactitol g. Using prolin as an osmo-protector, the concentration of produced hexanoic acid increased to 5~6 g/L.

In this study, the repeated batch production of bioethanol from sucrose by self-flocculating yeast was conducted for the industrial production of bioethanol from sugary biomass. With recycling volume of 5%, the initial cell concentration of 30.1 g-dry cell weight/L was accumulated after 5 batches. For the repeated batch production of bioethanol from sucrose, the optimal fermentation medium was found to be 1 g/L urea, 1 g/L K2HPO4, 0.25 g/L MgSO4·7H2O. In this condition, the final ethanol concentration, the theoretical ethanol yield, and the volumetric productivity were attained to 74.6 g/L, 92.3%, and 6.21 g/L·h, respectively. Through the repeated batch fermentation, the initial cell concentration increased to approximately 5 times more than that of the batch fermentation, and the preculture system could be eliminated in the whole processes of ethanol production. Moreover, repeated batch fermentation using self-flocculating yeast was found to be superior in ethanol production not only on the non-sterilized condition of fermentation medium but also on the use condition of industrial water containing bacteria of 1200 CFU/ml.

Biological denitrification among various denitrification methods is the most widely accepted process because of its economical and environmental advantages. And bioelectrode has been studied. The bioelectrodes have some troubles–electron transfer, cost, and durability. In this study, Ochrobactrum anthropi SY509 was used as biocatalyst and the novel bioelectrode was made of silicone and graphite mixture. And in the bioelectrode, the electrons were t ransferred directly to the immobilized biocatalyst. And the graphite powder was coated by biocatalyst for increasing the electron transfer efficiency. Using this electrode, high denitrification efficiency and durability was obtained.

Lignocellulosic biomass is one of the most potential candidates for biogas and bioethanol production. Since structural features, such as high lignin content, crystallinity, and acetyl content make lignocelluloses recalcitrant to enzymatic degradation, its pretreatment is a key process. Pleurotus ostreatus, one of the white-rot fungi, was selected for biological pretreatment because it can modify the structural features for the better enzymatic digestibility with a capacity to remove lignin from lignocelluloses. The selective lignin-degrading ability of Pleurotus ostreatus comes from the ligninase system which is mainly composed of lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase. Nitrogen or carbon limitation induces lignin biodegradation and ligninase production, whereas carbon and nitrogen limitation also cause a loss of mycelia dry weight and low production of extracellular proteins including enzymes. To obtain maximal protein production without carbon catabolite repression, the optimized carbon and nitrogen supplies should be considered. Using the central composite design (CCD) with interactive variables of glucose (as C source) and ammonium sulfate (as N source), the optimization of C/N ratio for the biological pretreatment by Pleurotus ostreatus was investigated in this study.

Klebsiella pneumonia DSM 4799 can convert glycerol to 1,3-propanediol, which is a basic ingredient for polytrimethylene terephthalate (PTT), lubricant, solvents, and other end uses. In this study, inhibitory effect of initial concentration of 1,3-propandiol(1,3-PD) on 1,3-PD production was investigated. The initial concentration of 1,3-PD affected the growth and 1,3-PD production of K. pneumonia DSM 4799. With glycerol 20 g/L, the growth and the glycerol consumption rate were dramatically decreased with an initial 1,3-PD concentration higher than 40 g/L. When 1,3-PD was added at the initial concentration of 0, 20, 40, and 60 g/L, the 1,3-PD production was 7.9, 5.9, 5.3, and 0 g/L, respectively, after 12 hrs of cultivation. There was little 1,3-PD production with more than 60 g/L 1,3-PD. Also, the production of other products such as 2,3-butanediol, ethanol, and bio-gas decreased as initial 1,3-PD concentration increased. Unlike those products, acetic acid was produced more with 40 and 60 g/L 1,3-PD than with 0 and 20 g/L, implying that inhibited cells might produce more acetic acid due to the change in metabolism.

Optimization of sea algae acid saccharification conditions have been researched in this study.To determine kind of acid optimized for acid saccharification, sea algae has been acid saccharificated and compared glucose yield using nitric acid, sulfuric acid, hydrochloric acid and phosphoric acid in identical condition. Afterwards, using determined acid, it has been researched with variable conditions of acid concentration, reaction temperature, reaction time with RSM (Response Surface Methodology) to analyze sea algae acid saccharification conditions. Also, theoretical optimum acid saccharification conditions have been obtained from substituting acid saccharification conditions with RSM (Response Surface Methodology) to analyze into glucose yield follows in the solid and liquid ratio of reaction material and acid saccharification conditions yield experiment has been researched from this study to confirm theoretical conditions. Finally, decomposition behaviors follows in reaction condition have been confirmed defining over decomposed HMF(HydroxyMethul Fulfural), furfural, fructose and etc’s products.

Klebsiella pneumoniae is an excellent 1,3-PD producer, but the simultaneous production of many byproducts (2,3-butandiol, Ethanol, Succinic acid, etc.) have greatly reduced the fermentation efficiency of 1,3-PD. Hence, to eliminate the by-products synthesis, the oxidative branch of glycerol metabolism was inactivated by deleting from the chromosomal DNA the genes involved in the synthesis of by-products. The microbial production of 1,3-PD by recombinant K. pneumoniae was investigated in a two-step fermentation process. The first step, include the growth of the cells in LB broth, till absorbance at 600nm reached 1.5~2.0. The second step, involved the glycerol feeding at this stage, using this strategy glycerol was rapidly converted to 1,3-PD. In the present study, we attempted to statistically optimize the two-step fermentation conditions for 1,3-PD by the engineered strain of K. pneumoniae using response surface methodology (RSM) based on a 25 factorial central composite design (CCD) (1). The optimum conditions for two-step fermentation were: 6.5 pH; 0.7 vvm aeration volume; 25.0 g/L glycerol concentration; 15.0 h fermentation time and 35.74°C temperature. The experimental production of 1,3-PD was 11.29 g/L, which was in close agreement with the model prediction. In this results, the productivity of 1,3-PD and the yield of 1,3-PD relative to glycerol reached 0.63 g/Lh and 0.66 mol/mol, respectively, which were about 2 fold higher than those of one-step fermentation.

reserves. There is a need of increased efforts required for the development of new alternative energy sources, which replace crude oil, with less cost and eco-friendly approaches. Bioethanol production from crop wastes is one of the major work progresses by various countries to meet their energy consumption in the last two decades. Delignification is the major problem in using biomass. We have compared acid, alkali and microwave treatment for delignification of rice straw. Alkaline treatment is found to be the most effective delignification process among those studied. Cellulases play a major role in the saccharification of biomass. Based on the preferential pretreatment approach, the suitable cellulase exploration produce authentic results. We have isolated a novel Bacillus amyloliquefaciens K5 strain producing a slightly alkaline pH stable cellulase. This enzyme found to be useful in reducing sugar production from alkali pretreated biomass fermentation. We have obtained a 44% saccharification by initial weight using the commercial Novozyme cellulase on alkali pretreated ricestraw. The results of various pretreatments for rice straw delignification, saccharification with commercial cellulases, media optimization using pretreated rice straw as a carbon source for higher cellulase production by Bacillus amyloliquefaciens K5 will be presented.

After industrial cultivation for vitamin B2, untreated wastewater may result in stench and eutrophication of sea if it would be dumped in the sea because of high contents carbon, nitrogen and sulfur elements in wastewater. In order to recycle the wastewater, therefore, the removal of the stench is top priority.We employed Effective Microorganisms (EM) for deodorization because EM is widely applied to deodorize H2S and NH3. In this study, experiments were performed in 1L fermenter under anaerobic condition with N2 gas. We supplied sugar as additional carbon source. We measured the concentration of H2S in culture broth, cell growth and pH. H2S was measured by titrimetric method after gathering AVS(acid volatile sulfide). Cell growth was observed by plate counting method for viable cell count. Because EM contains many other genera, we employed selective pressure such as cycloheximide, erythromycin, rogosa media for each cell counting. Consequently, the H2S concentration of EM treated group was reduced dramatically compared to control group. This study revealed the potential application of EM to treatment of stench in wastewater.

A novel anaerobic bacteria strain was used for the production of biohydrogen with food wastes. Among a few hydrogen producing strains which were isolated from natural environment, one of the isolate, designated FH2072, exhibited the highest hydrogen production. This isolate was identified to belong to Clostridium aurantibutyricum using 16S rDNA analysis. Hydrogen production by C. aurantibutyricum FH2072 with a dried food waste product (FWP), obtained from EService Corp., was surveyed in batch culture tests. C. aurantibutyricum FH2072 produced H2 using the FWP under anaerobic conditions regardless of enzyme hydrolysis of FWP. In a medium containing 3% (w/v) hydrolyzed FWP, C. aurantibutyricum FH2072 exhibited the maximum hydrogen production rate of 56 ml H2 l-1h-1 and the total hydrogen production of 1848 ml H2 l-1.

Biofuel cells are a specific type of fuel cell which uses biocatalysts such as enzymes and microorganisms to gain electric energy from chemical energy. They do not need high pressure and temperature for their working. The most potential application of enzymatic biofuel cells, which consist of a bioanode which oxidizes glucose to glucolactone and a biocathode which reduces oxygen to water, is for implantable power sources such as microscale cells implanted in blood vessels. As a bioanode for enzymatic fuel cells, we selected the following modified electrode. Glucose oxidase (GOx) was entrapped in a matrix of multi-walled carbon nanotubes (MWCNTs) and chitosan on a glassy carbon electrode. MWCNTs selectively wrapped by a water-soluble, environmentally-friendly, biocompatible polymer chitosan were employed for the construction of a bioelectrochemical platform for the direct electron transfer (DET) of GOx. They are also expected to act as an electron transfer promoter. Using cyclic voltammetry, we checked that the DET between GOx and anode took place in a phosphate buffer solution (100 mM, pH 7) saturated with argon as well as with oxygen. The experimental results demonstrated that the immobilized GOx retained its catalytic activity towards the oxidation of glucose under the aerobic conditions.