• Journal of Microbiology and Biotechnology
• /
• v.26 no.8
• /
• pp.1348-1357
• /
• 2016

An enzymatic reaction system was developed and optimized for bioconversion of resveratrol from glucose. Liquid enzyme extracts were prepared from Alternaria sp. MG1, an endophytic fungus from grape, and used directly or after immobilization with sodium alginate. When the enzyme solution was used, efficient production of resveratrol was found within 120 min in a manner that was pH-, reaction time-, enzyme amount-, substrate type-, and substrate concentration-dependent. After the optimization experiments using the response surface methodology, the highest value of resveratrol production (224.40 μg/l) was found under the conditions of pH 6.84, 0.35 g/l glucose, 0.02 mg/l coenzyme A, and 0.02 mg/l ATP. Immobilized enzyme extracts could keep high production of resveratrol during recycling use for two to five times. The developed system indicated a potential approach to resveratrol biosynthesis independent of plants and fungal cell growth, and provided a possible way to produce resveratrol within 2 h, the shortest period needed for biosynthesis of resveratrol so far.

• Journal of Korean Society of Water and Wastewater
• /
• v.23 no.2
• /
• pp.175-180
• /
• 2009

This study was conducted to improve biological degradation efficiency of toluene as a model volatile organic compound (VOC) using yeast Candida tropicalis and to suggest an effective method for bioreactor operation. The yeast strain was immobilized with polyethylene glycol (PEG), alginate, and powdered activated carbon (PAC). The yeast-immobilized polymer media were used as fluidized materials in an airlift bioreactor. Polymer media without PAC were also made and operated in another airlift bioreactor. The two bioreactors showed toluene removal efficiencies ranging 80-96% at loading rates of $10-35 g/m^3-hr$, and the bioreactor containing the polymer media with PAC achieved higher removal efficiency. Protein contents in the liquid phase showed that the bioreactor using the yeast-immobilized polymer media with PAC had a higher rate of microbial growth initially than that without PAC. In addition, the microbial growth rate inside of the polymer media with PAC was five times higher than that without PAC. Consequently, the polymer media containing the yeast strain and PAC could enhance removal efficiencies for VOCs, and the immobilization method improve microbial activity and stability for a long-term operation of biological systems.

• The Korean Journal of Food And Nutrition
• /
• v.13 no.4
• /
• pp.328-333
• /
• 2000

The spore of Aspergillus niger KCTC-6144 was immobilized on alginate gel beads. When pumpkin powder was used with glucose for a medium of citric acid fermentation by Aspergillus niger beads, the beaded Aspergilus niger grew up inside the bead and mycelia penetrated through the pore of the bead membrane. The bead size became largely from 2.0∼2.5mm to 6∼8mm after growing at 30$\^{C}$ for 4 days. Studies of optimum culture conditions on citric acid fermentation using Aspergillus niger beads on pumpkin medium (pumpkin powder 1% +glucose 7%, pH 6.0) were carried out in submerged cultures on 250m1 Erlenmeyer flask. As a result, it was found that to reinforce 12% as carbon source was good for citric acid production and that 1% pumpkin powder was good as nitrogen and mineral source in orbital shaker (150rpm) at 30$\^{C}$ for 5 days. The optimum initial pH on citric acid production was pH 6.0 and it was found that 100 beads of immobilized Aspergillus niger was adequate for citric acid production in a 250ml Erlenmeyer flask containing 50m3 of pumpkin medium solution with orbital shaker at 30$\^{C}$ for 5 days. We also found that maximal production of citric acid was 23.5g/ℓ at optimal condition (at 30$\^{C}$ for 5 days, pH 6.0, and 100 beads and medium containing 1% pumpkin powder plus 12% glucose).

• Microbiology and Biotechnology Letters
• /
• v.44 no.4
• /
• pp.504-511
• /
• 2016

Lactulose, a synthetic disaccharide, has received increasing interest because of its role as a prebiotic that can increase the proliferation of Bifidobacterium and Lactobacillus spp. and enhance the absorption of calcium and magnesium. While the industrial production of lactulose is still mainly achieved by the chemical isomerization of lactose in alkaline media, this process has drawbacks including the need to remove catalysts and by-products, as well as high energy requirements. Recently, the use of cellobiose 2-epimerase (CE) has been considered an interesting alternative for industrial lactulose production. In this study, to develop a process for enzymatic lactulose production using CE, we screened improved mutant enzymes ($CS-H^RC^E$) from a library generated by an error-prone PCR technique. The thermostability of one mutant was enhanced, conferring stability up to $75^{\circ}C$, and its lactulose conversion yield was increased by 1.3-fold compared with that of wild-type CE. Using a recombinant Escherichia coli strain harboring a CS35 $H^RC^E$-expressing plasmid, we prepared cell beads immobilized on a Ca-alginate substrate and optimized their reaction conditions. In a batch reaction with 200 g/l lactose solution and the immobilized cell beads, lactose was converted into lactulose with a conversion yield of 43% in 2 h. In a repeated 38-plex batch reaction, the immobilized cell beads were relatively stable, and 80% of the original enzyme activity was retained after 4 cycles. In conclusion, we developed a reasonable method for lactulose production by immobilizing cells expressing thermostable CE. Further development is required to apply this approach at an industrial scale.

• Journal of Korean Society of Environmental Engineers
• /
• v.31 no.8
• /
• pp.603-610
• /
• 2009

This research was performed to improve removal efficiency of toluene and methyl ethyl ketone (MEK) using Candida tropicalis, one of the yeast species. An airlift loop bioreactor (ALB) was employed to enhance the capability of mass transfer for toluene and MEK from the gas phase to the liquid, microbial phase. Polymer gel media made from PAC, alginate and PEG was applied for the effective immobilization of the yeast strain on the polymer gel media. The experimental results indicated that the mass transfer coefficient of toluene without polymer gel media was 1.29 $min^{-1}$ at a gas retention time of 15 sec, whereas the KLa value for toluene was increased to 4.07 $min^{-1}$ by adding the media, confirming the enhanced mass transfer of volatile organic compounds between the gas and liquid phases. The removal efficiency of toluene and MEK by using yeast-immobilized polymer gel media in the ALB was greater than 80% at different pollutant loading rates (5, 10, 19 and 37 g/$m^3$/hr for toluene, 4.5, 8.9, 17.8 and 35.1 g/$m^3$/hr for MEK). In addition, an elimination capacity test conducted by changing inlet loading rates stepwise demonstrated that maximum elimination capacities for toluene and MEK were 70.4 and 56.4 g/$m^3$/hr, respectively.

• Korean Journal of Food Science and Technology
• /
• v.23 no.5
• /
• pp.561-567
• /
• 1991

This study was designed to investigate the productivity of L-methionine by the method of protoplast fusion between Brevibacterium flavum ATCC 14067 and Corynebacterium glutamicm ATCC 13032, and then L-methionine production was performed to continuous fermentation using the immobilized fusant cells. Mutants B. flavum K 104($thr\;met\;Km^{r}\;Et^{r}\;Sm^{r}\;Tm^{r}\;as\;genetic\;marker$) and C. glutamicum B 70($thr\;Hos\;Km^{r}\;Et^{r}\;Sm^{r}\;Tm^{r}as\;genetic\;marker$) were isolated by MNNG treatment. On the other hand, protoplast of mutants were formed to treat with lysis solution containing $500{\mu}g/ml$ of lysozyme. The ratios of protoplast formation and regeneration were 99% and $64{\sim}66%$ respectively. Fusion frequency between B. flavum K 104 and C. glutamicum B 70 showed the $3.5{\times}10^{5}$ in the 35% polyethylene glycol(PEG6000) containing 3% PVP solution. The productivity of L-methionine by fusant BFCG 37 immobilized with sodium alginate was 0.89 g/l the batch fermentation and was $18.75mg/^{1}hr\;^{1}$ on the continuous fermentation at $30^{\circ}C$ for 72 hr.

• Korean Journal of Food Science and Technology
• /
• v.22 no.6
• /
• pp.611-617
• /
• 1990

This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

• Journal of Environmental Impact Assessment
• /
• v.30 no.4
• /
• pp.247-257
• /
• 2021

In this work, Iron Nano-Particles Impregnated BioChar/bead (INPBC/bead) soil amendment was developed to increase biochar's reactivity to As in soil and preventing possible wind loss. Prior to preparation of INPBC/bead, INPBC was produced utilizing lignocellulosic biomass and Fe(III) solution in a hydrothermal method, followed by a calcination process. Then, the bead type amendment, INPBC/bead was produced by cross-linking reaction of alginate with INPBC. FT-IR, XRD, BET, and SEM-EDS analyses were utilized to characterize the as-synthesised materials. The particle size range of INPBC/bead was 1-4 mm, and different oxygen-containing functional groups and Fe3O4 crystalline phase were produced on the surface of INPBC/bead, according to the characterization results. The soil cultivation test was carried out in order to assess the stabilization performance of INPBC/bead utilizing As and Pb-contaminated soil obtained from an abandoned mining location in South Korea. After 4 weeks of culture, TCLP and SPLP extraction tests were performed to assess the stabilization efficacy of the amendment. The TCLP and SPLP findings revealed that raising the application ratio improved stabilizing efficiency. The As stabilization efficiency was determined to be 81.56 % based on SPLP test findings for a 5% in (w/w) INPBC/bead treatment, and the content of Pb in extracts was reduced to the limit of detection. According to the findings of this study, INPBC/bead that can maintain pH of origin soil and minimize wind loss might be a potential amendment for soil polluted with As and heavy metals.

• Microbiology and Biotechnology Letters
• /
• v.28 no.2
• /
• pp.117-123
• /
• 2000

The object of this work is to improve the maintenance of bioluminescence from stored Photobacterium phosphoreum in a view of developing continuous monitoring system for pollutants. The long-term experiments were performed to determine the effect of storage temperature and immobilization on the maintenance of bioluminescence and viability of P. phosphoreum. A naturally luminescent bacterium, P. phosphoreum was starved in 2.5% Nael solution at $20^{\circ}C$, $4^{\circ}C$, -$20^{\circ}C$ and -$70^{\circ}C$ for 30 days. In vivo luminescence was measured by luminometry, and total cell concentrations and concentrations of culturable and viable cells were determined by acridine orange staining, dilution plate counting, and direct viable counting, respectively. The bioluminescence emission from cells stored at 4De was maintained up to 10 days while those with starved cells at other temperature ranges decreased to background level within 3 days. In terms of viability of cells, concentrations of cells stored at $20^{\circ}C$ were rapidly decreased as a result of cell lysis, leading to a drop in culturable and viable counts while cells stored at $4^{\circ}C$ was shown viable but nonculturable state during starvation. With immobilized cells on strontium alginate, the bioluminescence showed higher maintenance than free cells and decreased with count number of nonculturable cells.

• Microbiology and Biotechnology Letters
• /
• v.48 no.1
• /
• pp.12-23
• /
• 2020

Edible films containing antimicrobial agents can be used as safe alternatives to preserve food products. Essential oils are well-recognized antimicrobials. However, their low water solubility, volatility and high sensitivity to oxygen and light limit their application in food preservation. These limitations could be overcome by embedding these essential oils in complexed product matrices exploiting the encapsulation efficiency of β-cyclodextrin. This study focused on the maximization of β-cyclodextrin production using cyclodextrin glucanotransferase (CGTase) and the evaluation of its encapsulation efficacy to fabricate edible antimicrobial films. Response surface methodology (RSM) was used to optimize CGTase production by Brevibacillus brevis AMI-2 isolated from mangrove sediments. This enzyme was partially purified using a starch adsorption method and entrapped in calcium alginate. Cyclodextrin produced by the immobilized enzyme was then confirmed using high performance thin layer chromatography, and its encapsulation efficiency was investigated. The clove oil/β-cyclodextrin inclusion complexes were prepared using the coprecipitation method, and incorporated into chitosan films, and subjected to antimicrobial testing. Results revealed that β-cyclodextrin was produced as a major product of the enzymatic reaction. In addition, the incorporation of clove oil/β-cyclodextrin inclusion complexes significantly increased the antimicrobial activity of chitosan films against Staphylococcus aureus, Staphylococcus epidermidis, Salmonella Typhimurium, Escherichia coli, and Candida albicans. In conclusion, B. brevis AMI-2 is a promising source for CGTase to synthesize β-cyclodextrin with considerable encapsulation efficiency. Further, the obtained results suggest that chitosan films containing clove oils encapsulated in β-cyclodextrin could serve as edible antimicrobial food-packaging materials to combat microbial contamination.