The biosynthesis of lovastatin, a cholesterol lowering agent formed by the filamentous fungus Aspergillus terreus, was examined in a 2.5 L jar fermenter. In batch bioreactor cultures conducted at various agitation rates, 400 rpm showed the best result in terms of lovastatin production. Notably, the effect of pH on lovastatin biosynthesis was found to be significant: when the pH was controlled at around 5.8 during the whole fermentation period, lovastatin concentration reached 598 mg/L, which is much hihger than the amounts obtained by pH-uncontrolled and pH 7.4-controlled fermentations. In addition, both L-histidine and L-tryptophan were observed to be favorable amino acids for the enhancement of lovastatin production when 6 g/L of the respective amino acids were supplemented at the beginning of the fermentation period. By further optimization of the production media and the physical environment, lovastatin production was increased to 836 mg/L (3.5 mg/L/hr) which is approximately 10 times higher than the productivity of the basic control culture.
The aim of this study attempted to verify the possibility of bioethanol production using wasted medium density fiberboard (wMDF). In order to produce bioethanol from wood cellulosic materials must be carried out the process of pretreatment, saccharification, fermentation and distillation. First, the wMDF was pretreated using sodium chlorite and pretreated wMDF was prepared to 8% slurry and then slurry was saccharified with the commercial enzyme (Cellic CTec3). The fermentable sugar and pH of saccharified substrate were about 5.5% glucose and 4.4, respectively. Herein we compared the results of ethanol yield according to the nutrients added or without addition to increase ethanol yield. Ethanol fermentation was finished in about 24 hours, but it was delayed in experimental group without nutrients. Ethanol content and fermentation ratio of the final fermented mash prepared by utilizing jar fermenter was 25.40 g/L and 86.64%, respectively. At this time, the maximum ethanol productivity was confirmed as 1.78 g/Lh (ethanol content 21.38 g/L, 12 h), and the overall ethanol productivity was 1.05 g/Lh (ethanol content 25.27 g/L, 24 h). Using fermented liquid we could produced bioethanol 95.37% by continuous distillator packed with copper element in laboratory scale. These results show that wMDF has a potential valuable for bioethanol production.
Pseudomonas sp. BK7, an alkalophile, displayed the highest growth and protease activity when grown in a fermenter which was controlled at a pH level of 9.0, and the enzyme production was significantly enhanced by the increase of agitation speed. Two forms of alkaline proteases (BK7-1 and BK7-2) were fractionated and purified to near homogeneity. Protease BK7-1 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B, DEAE-Sepharose, and Sephadex G-75 column chromatographies. The molecular weights of proteases BK7-1 and BK7-2 determined by gel filtration chromatography were 20,700 and 40,800, respectively. The $K_m$ value, isoelectric point, and optimum pH of protease BK7-1 were 2.55 mg/ml, 11.0, and 11.0, respectively, whereas those of protease BK7-2 were 1.57 mg/ml, 7.2, and 10.0, respectively. Both proteases were practically stable in the pH range of 5-11. The optimum temperatures for the activities of both protease BK7-1 and BK7-2 were $50^{\circ}C$ and $45^{\circ}C$, respectively. About 56% of the original protease BK7-2 activity remained after being treated at $50^{\circ}C$ for 30 min but protease BK7-1 was rapidly inactivated at above $25^{\circ}C$. Both proteases were completely inhibited by phenylmethane sulfonyl fluoride, a serine protease inhibitor. Protease BK7-2 was stable against EDTA, EGTA, STP, and detergents such as SDS and LAS, whereas protease BK7-1 was found to be unstable.
To improve the fermentation characteristics of the haploid starch-fermenting recombinant yeast strain K114/YIpMS$\Delta$R(LEU2/URA3) secreting both $\alpha$-amylase and glucoamylase was rare-mated with polyploid industrial yeast Saccharomyces sp. K35. The K35 strain had good fermentation-characteristics such as ethanol-tolerance, high temperature and sugar-tolerance, and high fermentation rate. Among the resulting 66 hybrids, the best strain RH51 was selected. The RH51 exhibited amylolytic activity of K114/YIpMS$\Delta$R(LEU2/URA3) as well as ethanol and sugar tolerance of K35. The optimum temperature of hybrid RH51 for starch fermentation was 34$\circ$C which was same as that of K35 but different from that (30$\circ$C) of K114/YIpMS$\Delta$R(LEU2/URA3). The optimum pH was 5.0. The optimum size of inoculum was 2% as the pellet (w/v) of yeast cells. The hybrid strain RH51 produced 7.0% ethanol (w/v) from 20% (w/v) soluble starch while K35 formed almost no ethanol, 0.3% (w/v). RH51 strain produced 7.5% (w/v) ethanol after 8 days in a 2.5 l fermenter containing 800 ml of 20% (w/v) soluble starch. The residual starch content in the fermentation medium was 1.68% (w/v), and therefore almost all the starch was fermented completely.
Kim, Dal-Soo;Rae, Cheol-Yong;Chun, Sam-Jae;Kim, Do-Hyung;Choi, Sung-Won;Choi, Kee-Hyun
The Plant Pathology Journal
/
v.21
no.4
/
pp.328-333
/
2005
Paenibacillus elgii SD17 (KCTC $10016BP^T$=NBRC $100335^T$) was recently reported as a new species. Based on its inhibitory activity to Thanatephorus cucumeris AG1-1, strain SD17 was further evaluated for its potential as a biocontrol agent against soil-borne diseases of turf grasses in Korea. P. elgii SD17 showed a broad spectrum of antimicrobial activity in vitro test and suppressed development of turf grass diseases; Pythium blight caused by Pythium aphanidermatum and brown patch caused by T. cucumeris AG1-1 on creeping bentgrass (Agrostis palustris) in the growth chamber tests. Under a condition for massive culture in a 5,000 L fermenter, P. elgii SD17 reached $6.4{\times}10^8$ spores/ml that resulted in approximately $1.0{\times}10^7$ cfu/g when formulated into a granule formulation (GR) using the whole culture broth instead of water. Using the GR formulation, biocontrol activity of P. elgii SD17 was confirmed. In the growth chamber tests, the GR formulation was effective against brown patch and Pythium blight with similar level of disease severity compared to each of the standard fungicides at the application rates of 10 g/$m^2$ or above. In the field tests, compared to each untreated control, the GR formulation also effectively controlled Pythium blight, brown patch and large patch at all the application rates of 5, 10 and 20 g/$m^2$, respectively, without significant response by the application rates. However its performance was inferior to each of the standard chemical fungicides. Based on these results, we consider this GR formulation of P. elgii SD17 as an effective biocontol agent to suppress Pythium blight, brown patch and large patch of turf grasses in Korea.
Bacterial lysates have become a common ingredient for natural health care. Lactic acid bacteria (LAB) could serve as potential candidates for lysate production: the lactic acids produced by LAB have been utilized for their moisturizing, antimicrobial, and rejuvenating effects, while other substances provide topical benefits and health effects for the skin. Our study aimed to obtain lysate from a LAB S. macedonicus MBF 10-2 through an optimized fermentation using the Response Surface Methodology. Strain MBF10-2 was cultivated in a 2L fermenter tank in de Man Rogosa and Sharpe (MRS) medium and in plant-based peptone modified MRS, i.e. Soy-peptone and Vegitone. The duration and the medium composition (dextrose and soy peptone or proteose peptone) were adjusted to obtain an optimum production of cell lysate. Central Composite Design was employed for Design Expert 7.0.0 by adjusting 3 factors: dextrose (1%, 1.5%, 2%, 2.5%, 3%), soy or proteose peptone (0.5%, 0.75%, 1%, 1.25% and 1.5%), and duration of fermentation (8, 10, 12 14, and 16 h for MRS-Soy peptone and 15, 17, 19, 21, and 23 h for MRS Vegitone). Bacteriocin-Like Inhibitor Substance activity of lysate and pH were used as indicators. The optimum condition for lysate production using MRS Soy Peptone and Vegitone are as follows: dextrose concentration 2.5%, plant-based peptone 1.25%, while optimum fermentation duration were 11.18 h (MRS Soy Peptone) and 17 h (MRS Vegitone) with a starter concentration of 10% at $OD_{600nm}$$0.2{\pm}0.05$. However, the standard MRS medium produced better quality lysate compared to MRS plant-based peptones.
Journal of Korean Society of Environmental Engineers
/
v.27
no.1
/
pp.85-92
/
2005
The performance of a novel fermentation process, adopting a sludge blanket type configuration for higher hydrolysis/acidogenesis of the municipal primary sludge, was investigated under batch and semi-continuous conditions with various pH and temperature conditions. This acid elutriation slurry reactor provided higher system performance with a short HRT (5 days) and higher acidogenic effluent quality under pH 9 and thermophilic ($55^{\circ}C$) conditions. The hydrolysis of the sludge was revealed to be significantly dependent on seasonal effects for sludge characteristics but with little impact on acidogenesis. Based on the rainy season at the optimum conditions, VFA production and recovery fraction ($VFA_{COD}/COD$) were $0.18\;g\;VFA_{COD}\;g^{-1}\;VSS_{COD}$ and 63%. As byproducts, nitrogen and phosphorus releasing were $0.006\;g\;N\;g^{-1}\;VSS_{COD}$ and $0.003\;g\;P\;g^{-1}\;VSS_{COD}$, respectively. For the mass balance in a full-scale plant($Q=158,880\;m^3\;day^{-1}$) based on the rainy season, the VFA and non-VFA(as COD) production were $3,110\;kg\;VFA_{COD}\;day^{-1}$ and $1,800\;kg\;COD\;day^{-1}$, resulting in an increase of organics of $31\;mg\;COD\;L^{-1}$ and $20\;mg\;VFA_{COD}\;L^{-1}$ and nutrients of $0.7\;mg\;N\;L^{-1}$ and $0.3\;mg\;P\;L^{-1}$ in the influent sewage. The economical benefit from this process application was estimated to be about $67 per $1,000m^3$ of sewage except for energy requirements and also, better benefits can be expected during the dry season. Also, the results revealed that the process has various additional advantages such as pathogen-free stabilized solids production, excellent solids control and economical benefits.
${\gamma}-Aminobutyric$ acid (GABA) producing lactic acid bacteria, Lactobacillus acidophilus RMK567 was cultivated in 50 L of sterilized MRS broth using a fermenter at $40^{\circ}C$ for 24 h. The cell number was increased to $10.04{\pm}0.13$ Log CFU/mL with a growth rate constant (k) of 0.454 generation/h and a generation time (g) of 2.303 h after a lapse of a lag phase (L) of 5.16 h. A total of 487 g of cell paste with 40.5% moisture was harvested with viable cell number of 12.48 Log CFU/g cell paste. The cell pastes after preparation with glycerol, glucose, and polydextrose as cryo-protectants were lyophilized under a vacuum of 84 m torr. A total of 408 g of freeze dried (FD) cell powders were mixed with a commercial strain of Streptococcus thermophilus to prepare of three types FD starter cultures with the viable cell numbers of 12.42 (FDA-GY), 12.60 (FDBGG) and 12.91 (FDC-GP) Log CFU/g. During preservation the FD cultures at -$18^{\circ}C$, the cell viability of the FD starter cultures were rapidly dropped to below 3.24% of the day of storage. No significant difference was found in the cell viabilities among three types of FD starters cultures, but significant difference (p<0.01) was found in storage periods. Yoghurts fermented through FD starter culture of L. acidophilus RMK567 were determined to contain $155.16{\pm}8.53$ ppm, $243.82{\pm}4.27$ ppm, and $198.64{\pm}23.46$ ppm of GABA, respectively. This study shows that GABA production activity of L. acidophilus RMK567 is not affected during the freeze drying process and would be available for commercial production of yoghurt containing high GABA content.
Journal of Korean Society of Environmental Engineers
/
v.28
no.8
/
pp.866-871
/
2006
The nutrient recovery in phosphate crystallization process was investigated by using laboratory scale uptlow reactors, adopting sequencing batch type configuration. The industrial waste lime was used as potential cation source with magnesium salt($MgCl_2$) as control. The research was focused on its successful application in a novel integrated sludge treatment process, which is comprised of a high performance fermenter followed by a crystallization reactor. In the struvite precipitation test using synthetic wastewater first, which has the similar characteristics with the real fermentation effluent, the considerable nutrient removal(about 60%) in both ammonia and phosphate was observed within $0.5{\sim}1$ hr of retention time. The results also revealed that a minor amount(<5%) of ammonia stripping naturally occurred due to the alkaline(pH 9) characteristic in feed substrate. Stripping of $CO_2$ by air did not increase the struvite precipitation rate but it led to increased ammonia removal. In the second experiment using the fermentation effluent, the optimal dosage of magnesium salt for struvite precipitation was 0.86 g Mg $g^{-1}$ P, similar to the mass ratio of the struvite. The optimal dosage of waste lime was 0.3 g $L^{-1}$, resulting in 80% of $NH_4-N$ and 41% of $PO_4-P$ removal, at about 3 hrs of retention time. In the microscopic analysis, amorphous crystals were mainly observed in the settled solids with waste lime but prism-like crystals were observed with magnesium salt. Based on mass balance analysis for an integrated sludge treatment process(fermenter followed by crystallization reactor) for full-scale application(treatment capacity Q=158,880 $m^3\;d^{-1}$), nutrient recycle loading from the crystallization reactor effluent to the main liquid stream would be significantly reduced(0.13 g N and 0.19 g P per $m^3$ of wastewater, respectively). The results of the experiment reveal therefore that the reuse of waste lime, already an industrial waste, in a nutrient recovery system has various advantages such as higher economical benefits and sustainable treatment of the industrial waste.
Bacillus polyfermenticus SCD, which is commonly called a 'Bisroot' strain, has been appropriately used for the treatment of long-term intestinal disorders, since the live strains, in the form of active endospores, can successfully reach the target intestine. Goal of this study was to develop an industrial medium for growth and sporulation of B. polyfermenticus SCD. From the results of effect of mixed carbon sources on growth and sporulation of B. polyfermenticus SCD, glucose 2% and starch 2% was particularly found to be the most effective for the maximum number of spore production, resulting in spore cells of $4.3{\times}10^9\;spores/mL$ with a sporulation yield of 91%. For the effect of nitrogen sources, the maximum spore cells of $5.7{\times}10^9\;spores/mL$ of B. polyfermenticus SCD with a sporulation yield of 97% was obtained when B. polyfermenticus SCD was cultivated in an optimum nitrogen source medium containing 5% soybean flour. A medium involving proper phosphate salt yielded the maximum number of a spore cells of $6.0{\times}10^9\;spores/mL$ with a sporulation yield of 95%. Finally, the efficacy of an industrial medium (KH5 medium) on growth and sporulation of B. polyfermenticus SCD was investigated in jar fermenter. The higher number of viable cells $(3.3{\times}10^{10}\;cells/mL)$ and spore cells $(3.0{\times}10^{10}\;spores/mL)$ were obtained in 5 L fermenter when compared with a 500 mL baffle flask cultivation. Thus, KH5 medium developed in this study shows promise as an industrial medium because of higher cells and sporulation yield.
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