• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.491-505
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• 2020

A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.501-509
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• 2021

The increasing demand for baked products has given a boost to research on isolation and selection of novel yeast strains with improved leavening activity. Twelve sourdough samples were collected from several localities of the Fez region in Morocco. The pH and total titratable acidity (TTA) values of these samples varied from 3.03-4.63 and 14-17.5 ml of 0.1 N NaOH/10 g of sourdough, respectively, while yeast counts ranged from 5.3 6.77 Log CFU/g. Thirty-two yeast isolates were obtained and evaluated for their leavening ability. Out of all isolates, four yeasts molecularly identified as Saccharomyces cerevisiae (three strains) and Kluyveromyces marxianus (one strain) showed highest specific volumes of 4.69, 4.55, 4.35 and 4.1 cm³/g, respectively. These strains were further assessed for their tolerance to high concentrations of salt, sugar, elevated temperatures, and low pH conditions. K. marxianus showed higher resistance than the S. cerevisiae. Thus, Moroccan sourdoughs harbor technologically relevant yeasts that could be used as potential starters for bread preparation.

• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.99-109
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• 2022

This study is the first report on production and characterization of the enzyme from an Ornithinibacillus species. A 4.2-fold increase in the extracellular protease (called L9^T) production from Ornithinibacillus caprae L9^T was achieved through the one-factor-at-a-time approach and response surface methodological optimization. L9^T protease exhibited a unique protein band with a mass of 25.9 kDa upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This novel protease was active over a range of pH (4-13), temperatures (30-80℃) and salt concentrations (0-220 g/l), with the maximal activity observed at pH 7, 70℃ and 20 g/l NaCl. Proteolytic activity was upgraded in the presence of Ag⁺, Ca²⁺ and Sr²⁺, but was totally suppressed by 5 mM phenylmethylsulfonyl fluoride, which suggests that this enzyme belongs to the serine protease family. L9^T protease was resistant to certain common organic solvents and surfactants; particularly, 5 mM Tween 20 and Tween 80 improved the activity by 63 and 15%, respectively. More importantly, L9^T protease was found to be effective in dehairing of goatskins, cowhides and rabbit-skins without damaging the collagen fibers. These properties confirm the feasibility of L9^T protease in industrial applications, especially in leather processing.

• Journal of People, Plants, and Environment
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• v.23 no.4
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• pp.455-464
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• 2020

Background and objective: This study was conducted to examine changes in the composition and physiological activity of Gardenia Fructus after being roasted. Methods: The antioxidant, anti-inflammatory and antibacterial activity of Gardenia Fructus was evaluated using the Gardenia Fructus (GF) and roasted Gardenia Fructus (RGF) ethanol extracts, and their components were analyzed through HPLC. Results: As a result, it was confirmed that the content of gardenoside and geniposide decreased and the content of genipin increased when GF was roasted. The total content of polyphenols was 54.5 ± 2.18 mg gallic acid equivalents (GAE) per gram of the GF extract and 69.6 ± 0.36 mg GAE per gram of the RGF extract. As a result of evaluating 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, both the GF and RGF extracts showed the similar activity to ascorbic acid at the concentrations of 1 mg/mL or higher. In RAW 264.7 macrophages stimulated by lipopolysaccharides (LPS), the RGF extract showed a higher effect of reducing NO production, and significantly reduced the expression of an inflammatory cytokine, IL-6. As a result of evaluating the antimicrobial activity, the RGF extract showed higher antimicrobial activity against Escherichia coli and Bacillus subtilis. In the dextran sulfate sodium salt (DSS) induced inflammatory bowel disease mouse model, the RGF extract reduced the weight of the spleen, and both the GF and RGF extracts reduced the number of bacteria in the colon. Conclusion: Therefore, it has been confirmed through this study that roasting at a high temperature changes the main components of the GF extract and increases its biological activity. The RGF extract is expected to be used as a natural material with antioxidant, anti-inflammatory and antibacterial effects.

• New & Renewable Energy
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• v.19 no.2
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• pp.31-39
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• 2023

Salinity gradient energy can be generated from a mixture of water streams with different salt concentrations by using reverse electrodialysis (RED). In this study, we evaluated the effect of stack size and number of cell pairs on the energy efficiency and specific energy of the RED process. Additionally, we studied the prementioned parameters to maximize the power density of RED. The performance of the RED stack which used a patterned ion exchange membrane, was evaluated as a function of stack size and feed flow rate. Moreover, it was noted that an increase in stack size increased the ion movement through the ion exchange membrane. Furthermore, an increase in feed flow rate led to a reduction in the concentration variation, resulting in an increase in OCV and power density. The energy efficiency and specific energy for 100 cells in the 10 × 10 cm² stack were the highest at 12% and 0.05 kWh/m³, respectively, while the power density from 0.33 cm/s to 5 × 5 cm² stack was the highest at 0.53 W/m². The study showed that the RED performance can be improved by altering the size of the stack and the number of cell pairs, thereby positively affecting energy efficiency and specific energy.

• Proceedings of the Microbiological Society of Korea Conference
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• 2001.11a
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• pp.39-45
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• 2001

Extremophiles are unique microorganisms that are adapted to survive in ecological niches such as high or low temperatures, extremes of pH, high salt concentrations and high pressure. These unusual microorganisms have unique biochemical features which can be exploited for use in the biotechnological industries. Due to the high biodiversity of extremophilic archaea and bacteria and their existence in various biotopes a variety of biocatalysts with different physicochemical properties have been discovered. The extreme molecular stability of their enzymes, membranes and the synthesis of unique organic compounds and polymers make extremophiles interesting candidates for basic and applied research. Some of the enzymes from extremophiles, especially hyperthermophilic marine microorganisms (growth above $85^{\circ}C$), have already been purified in our laboratory. These include the enzyme systems from Pyrococcus, Pyrodictium, Thermococcus and Thermotoga sp. that are involved in polysacharide modification and protein bioconversion. Only recently, the genome of the thermoalkaliphilic strain. Anaerobranca gottschalkii has been completely sequenced providing a unique resource of novel biocatalysts that are active at high temperature and pH. The gene encoding the branching enzyme from this organism was cloned and expressed in a mesophilic host and finally characterized. A novel glucoamylase was purified from an aerobic archaeon which shows optimal activity at $90^{\circ}C$ and pH 2.0. This thermoacidophilic archaeon Picrophilus oshimae grows optimally at pH 0.7 and $60^{\circ}C$. Furthermore, we were able to detect thermoactive proteases from two anaerobic isolates which are able to hydrolyze feather keratin completely at $80^{\circ}C$ forming amino acids and peptides. In addition, new marine psychrophilic isolates will be presented that are able to secrete enzymes such as lipases, proteases and amylases possessing high activity below the freezing point of water.

• Journal of the Korean Society of Food Culture
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• v.23 no.2
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• pp.235-242
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• 2008

In this study, the physicochemical and microbiological properties of market-available skate (Raja kenojei) kimchi were evaluated in comparison to long-term fermented Baechu kimchi. The comparative results for the skate (Raja kenojei) and Baechu kimchi products are as follows: salt concentrations = 3.40% and 3.95%, respectively; pH and acidity values = 4.01 and 3.90, and 0.65% and 0.82%, respectively; redox potential Eh values = 119.82 mV and 123.08 mV respectively; reducing sugars levels = 15.51 mg% and 13.23mg%, respectively; ascorbic acid levels = 24.21mg% and 22.18mg%, respectively; color L-values = 46.86 and 44.54, a-values = 15.46 and 12.28, and b-values = 29.94 and 28.36, respectively; hardness properties = 11.36 kgf and 10.23 kgf, respectively; hot water soluble pectin (HWSP) contents and sodium hexametaphosphate soluble pectin (NaSP) contents = 15.23% and 17.35%, and 32.51% and 29.64%, respectively; hydrochloric acid soluble pectin (HClSP) contents = 55.1% and 53.2%, respectively; total viable cell counts = $4.78{\times}10^8$ and $1.35{\times}10^8$, respectively; lactic acid bacteria counts = $5.18{\times}10^5$ and $1.32{\times}10^5$, respectively; and yeast levels = $8.52{\times}10^5$ and $5.23{\times}10^5$, respectively.

• Journal of Microbiology and Biotechnology
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• v.34 no.4
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• pp.930-939
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• 2024

Mushroom laccases play a crucial role in lignin depolymerization, one of the most critical challenges in lignin utilization. Importantly, laccases can utilize a wide range of substrates, such as toxicants and antibiotics. This study isolated a novel laccase, named HeLac4c, from endophytic white-rot fungi Hericium erinaceus mushrooms. The cDNAs for this enzyme were 1569 bp in length and encoded a protein of 523 amino acids, including a 20 amino-acid signal peptide. Active extracellular production of glycosylated laccases from Saccharomyces cerevisiae was successfully achieved by selecting an optimal translational fusion partner. We observed that 5 and 10 mM Ca²⁺, Zn²⁺, and K⁺ increased laccase activity, whereas 5 mM Fe²⁺ and Al³⁺ inhibited laccase activity. The laccase activity was inhibited by the addition of low concentrations of sodium azide and ⳑ-cysteine. The optimal pH for the 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt was 4.4. Guaiacylglycerol-β-guaiacyl ether, a lignin model compound, was polymerized by the HeLac4c enzyme. These results indicated that HeLac4c is a novel oxidase biocatalyst for the bioconversion of lignin into value-added products for environmental biotechnological applications.

• Tuberculosis and Respiratory Diseases
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• v.61 no.4
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• pp.374-383
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• 2006

Background: Ethyl pyruvate (EP) is a derivative of pyruvate that has recently been identified by both various in vitro and in vivo studies to have antioxidant and anti-inflammatory effects. The aim of this study was to determine the effect of EP on lipopolysaccharide (LPS)-induced acute lung injury (ALI). Methods: 5 weeks old, male BALB/c mice were used. ALI was induced by an intratracheal instillation of LPS 0.5mg/Kg/$50{\mu}L$ of saline. The mice were divided into the control, LPS, EP+LPS, and LPS+EP groups. In the control group, balanced salt solution was injected intraperitoneally 30 minutes before or 9 hours after the intratracheal instillation of saline. In the LPS group, a balanced salt solution was also injected intraperitoneally 30 minutes before or 9 hours after instillation the LPS. In the EP+LPS group, 40mg/Kg of EP was injected 30 minutes before LPS instillation. In the LPS+EP group, 40mg/Kg of EP was injected 9 hours after LPS instillation. The TNF-$\alpha$ and IL-6 concentrations in the bronchoalveolar lavage fluid (BALF), and that of NF-$\kappa$B in the lung tissue were measured in the control, LPS and EP+LPS groups at 6 hours after instillation of saline or LPS, and the ALI score and myeloperoxidase (MPO) activity were measured in all four groups 24 and 48 hours after LPS instillation, respectively. Results: The TNF-$\alpha$ and IL-6 concentrations were significantly lower in the EP+LPS group than in the LPS group (p<0.05). The changes in the concentration of these inflammatory cytokines were strongly correlated with that of NF-$\kappa$B (p<0.01). The ALI scores were significantly lower in the EP+LPS and LPS+EP groups compared with the LPS group (p<0.05). In the EP+LPS group, the MPO activity was significantly lower than the LPS group (p=0.019). Conclusion: EP, either administered before or after LPS instillation, has protective effects against the pathogenesis of LPS-induced ALI. EP has potential theurapeutic effects on LPS-induced ALI.

• The Korean Journal of Physiology
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• v.20 no.2
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• pp.218-224
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• 1986

The characteristics of anion exchange with internal $HCO_3\;^{-}\;(or\;OH^-)$ was studied by determining the time course of hemolysis in isoosmotic ammonium salt solution in human erythrocytes. The effects of inhibitors, pH and temperature on the exchange between internal $HCO_3\;^-\;(or\;OH^-)$ and external $Cl^-$ were observed and the permeabilities of various organic and inorganic anions were also measured. The results were compared with data previously reported from the experiments using radioisotopes. The results are as follows; 1) SITS $H_2DIDS$ and furosemide inhibited the hemolysis of erythrocytes in isoosmotic $NH_4Cl$ solution in a dose·dependent manner, and the concentrations for lengthening twice the time for $half-hemloysis(t_{1/2})\;were\;2.3{\times}10^{-7},\;1.3{\times}10^{-7}\;and\;2.5{\times}10^{-5}M$, respectively. 2) Acetazolamide also shifted the time-dependent hemolytic curve to the right in a dose-dependent manner, and the concentrations for lengthening twice $t_{1/2}\;was\;2.4{\times}10^{-5}M$. 3) The time-dependent hemolysis was delayed by decreasing pH from 7.0 to 6.2, but w·as not affected by the change of pH in the range of 7.0 to 8.2. 4) The time for $half-hemloysis(t_{1/2})$ showed a temperature-dependency and Arrhenius plot exhibited a break point at $20^{\circ}C$. The apparent activation energy calculated from this plot was 18.1 kcal/mol between $2^{\circ}C-20^{\circ}C$ and 11.2 kcal/mol between $20^{\circ}C-37^{\circ}C$, respectively. 5) The apparent permeabilities of various inorganic anions based on $t_{1/2}$ were in the order of $Cl^->NO_{3}\;^->SCN^->SO_4\;^{2-}>SSO_3\;^{2-}>HPO_4\;^{2-}$. which was similar with the previous reports based on the experiment using radioisotopes. The results Obtained from this study are comparable with the previous data reported from the experiments using radioisotopes. This indicates that the hemolysis of erythrocytes in isoosmotic ammonium salt solution can be used as a simple and good method for the study of anion exchange in erythrocyte membrane.