Showing papers in "Biotechnology and Bioprocess Engineering in 2006"

Biodiesel production using a mixture of immobilized Rhizopus oryzae and Candida rugosa lipases

Abstract: Biodiesel conversion from soybean oil reached a maximum of 70% at 18 h using immobilized 1,3-specificRhizopus oryzae lipase alone. Biodiesel conversion failed to reach 20% after 30 h when immobilized nonspecificCandida rugosa lipase alone was used. To increase the biodiesel production yield, a mixture of immobilized 1,3-specificR. oryzae lipase and nonspecificC. rugosa lipase was used. Using this mixture a conversion of greater than 99% at 21 h was attained. When the stability of the immobilized lipases mixture was tested, biodiesel conversion was maintained at over 80% of its original conversion after 10 cycles.

Preparation and characterization of melamine-formaldehyde resin microcapsules containing fragrant oil

Abstract: In this study, melamine-formaldehyde microcapsules were prepared viain situ polymerization using peppermint oil as a core material, melamine-formaldehyde as the wall material, Tween 20 as the emulsifier, and poly (vinyl alcohol) as a protective colloid. The melamine-formaldehyde microcapsules prepared in this study were then evaluated with regard to their structures, thermal properties, particle size distributions, morphologies, and release behaviors.

Application of ionic liquids as mobile phase modifier in HPLC

Abstract: Ionic liquids are receiving an upsurge of interest as ‘green’ solvents; primarily as replacements for conventional media in chemical processes. Although ionic liquids are rather “young” modifier, their great potential in high-performance liquids chromatography (HPLC) has already been demonstrated. This review presents an overview of the applications of ionic liquids as mobile phase modifiers in HPLC.

Comparisons of Physical Properties of Bacterial Celluloses Produced in Different Culture Conditions Using Saccharified Food Wastes

Abstract: The saccharogenic liquid (SFW) obtained by the enzymatic saccharification of food wastes was used as a medium for production of bacterial cellulose (BC). The enzymatic saccharification of food wastes was carried out by the cultivation supernatant ofTrichoderma harziaum FJ1 culture.Acetobacter xylinum KJ1 was employed for the BC production culture. The physical properties, such as polymerization, crystallinity, Young's modulus, and tensile strength, of BCs produced by three culture methods: the static cultures using HS (Hestrin-Schramm) as a reference medium (A) or the SFW medium (B), the shaking culture (C) or the air circulation culture (D) using the SFW medium, were investigated. The degrees of polymerization of BCs produced under the different culture conditions (A∼D) showed 11000, 9500, 8500, and 9200, respectively. Young's modulus was 4.15, 5.0, 4.0, and 4.6 GPa, respectively. Tensile strength was 124, 200, 80, and 184 MPa, respectively. All of the BC had a form of cellulose I representing pure cellulose. In the case of the shaking culture, the degree of crystallinity was 51.2%, the lowest degree. Under the other culturing conditions, the trend should remain in the range of 89.7–84%. Overall, the physical properties of BC produced from SFW were similar to those of BC from HS medium, a commercial complex medium, and BC production by the air circulation culture mode brought more favorable results in terms of the physical properties and its ease of scale-up. Therefore, it is expected that a new BC production method, like air circulation culture using SFW, would contribute greatly to BC-related manufacturing.

Optimization of γ-polyglutamic acid production by Bacillus subtilis ZJU-7 using a surface-response methodology

Abstract: The components of the media used to elicit the biosynthesis of poly-γ-glutamic acid (γ-PGA) byBacillus subtilis ZJU-7 were investigated, particularly the carbon and nitrogen sources. Of the 7 carbon sources investigated, sucrose induced the highest rate of γ-PGA productivity; among the nitrogen sources, tryptone had the best effect for γ-PGA production. A 26−2 fractional factorial design was used to screen factors that influence γ-PGA production significantly, and a central composite design was finally adopted to formulate the optimal medium. γ-PGA productivity improved approximately 2-fold when the optimal medium was used compared with the original nonoptimized medium, and volumetric productivity reached a maximum of 58.2 g/L after a 24-h cultivation period.

Isolation of off-flavors and odors from tuna fish oil using supercritical carbon dioxide

Abstract: Off-flavors and unfavorable odors in tuna fish oil were successfully removed and identified using supercritical carbon dioxide extraction, while retaining variable compounds, polyunsaturated fatty acids such as EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid). Samples of oil were extracted in a 100 mL semi-batch stainless steel vessel under conditions which ranged from 8 to 20 MPa and 20 to 60°C with solvent (CO2) flows from 10 g/min. GC-MS was used to identify the main volatile components contributing to the off-flavors and odors which included 2-methyl-1-propanol, 2,4-hexadienal, cyclopropane, and octadiene. Analyses of oil extracted at 40°C, 20 MPa showed a 99.8% reduction in dimethyl disulfide. Other significant off-flavors identified were 2-methyl-butene, 3-hydroxy butanal and ethylbenzene.

One-Step Enzymatic Synthesis of Blue Pigments from Geniposide for Fabric Dyeing

Abstract: In this study, we describe a one-step chemoenzymatic reaction for the production of natural blue pigments, in which the geniposide from Gardenia extracts is transformed by glycosidases to genipin. Genipin is then allowed to react with amino acids, thereby generating a natural blue pigment. The β-glycosidases, most notably isolase (a variant of β-glucanase), recombinant β-glucosidase, Cellulase T, and amylases, were shown to hydrolyze geniposide to produce the desired pigments, whereas the α-glycosidases did not. Among the 20 tested amino acids, glycine and tyrosine were associated with the highest dye production yields. The optimal molar ratio of geniposide to glycine, two reactants relevant to pigment production, was unity. The natural blue pigments produced in this study were used to dye cotton, silk, and wool. The color yields of the pigments were determined to be significantly higher than those of other natural dyes. Furthermore, the color fastness properties of these dyes were fairly good, even in the absence of mordant.

Factors affecting the characteristics of melamine resin microcapsules containing fragrant oils

Abstract: Microcapsules containing fragrant oils as a core material were prepared byin situ polymerization, using melamine-formaldehyde prepolymer as the wall material. The several parameters, such as stirring times, stirring rates, emulsifier types, emulsifier concentrations, and the viscosity of the core materials, affect the characteristics of the microcapsules. These parameters were investigated by the analyses of microcapsule size, particle size distribution, and morphology. The average microcapsule size decreased with an increase in stirring time, stirring rate, emulsifier concentration, and viscosity of the core material. It was also found that poly(vinyl alcohol) as a protective colloid could enhance the stability of the melamine-formaldehyde microcapsules.

Eicosapentaenoic Acid (EPA) Biosynthetic Gene Cluster of Shewanella oneidensis MR-1: Cloning, Heterologous Expression, and Effects of Temperature and Glucose on the Production of EPA in Escherichia coli

Abstract: The putative EPA synthesis gene cluster was mined from the entire genome sequence ofShewanella oneidensis MR-1. The gene cluster encodes a PKS-like pathway that consists of six open reading frames (ORFs): ORFSO1602 (multi-domain beta-ketoacyl synthase, KS-MAT-4ACPs-KR), ORFSO 1600 (acyl transferase, AT), ORFSO 1599 (multi-domain beta-ketoacyl synthase, KS-CLF-DH-DH), ORFSO 1597 (enoyl reductase, ER), ORFSO 1604 (phosphopentetheine transferase, PPT), and ORFSO1603 (transcriptional regulator). In order to prove involvement of the PKS-like machinery in EPA synthesis, a 20.195-kb DNA fragment containing the genes was amplified fromS. oneidensis MR-1 by the long-PCR method. Its identity was confirmed by the methods of restriction enzyme site mapping and nested PCR of internal genesorfSO1597 andorfSO1604. The DNA fragment was cloned intoEscherichia coli using cosmid vector SuperCos1 to form pCosEPA. Synthesis of EPA was observed in fourE. coli clones harboring pCosEPA, of which the maximum yield was 0.689% of the total fatty acids in a clone designated 9704-23. The production yield of EPA in theE. coli clone was affected by cultivation temperature, showing maximum yield at 20°C and no production at 30°C or higher. In addition, production yield was inversely proportional to glucose concentration of the cultivation medium. From the above results, it was concluded that the PKS-like modules catalyze the synthesis of EPA. The synthetic process appears to be subject to regulatory mechanisms triggered by various environmental factors. This most likely occurs via the control of gene expression, protein stability, or enzyme activity.

Preservation of Coagulation Efficiency of Moringa oleifera, a Natural Coagulant

Abstract: In recent years, there has been an interest to useMoringa oleifera as the natural coagulant due to cost, associated health and environmental concerns of synthetic organic polymers and inorganic chemicals. However, it is known thatM. oleifera as the natural coagulant is highly biodegradable and has a very short shelf life. This research was carried out to investigate the effects of storage temperature, packaging methods, and freeze-drying on the preservation ofM. oleifera seeds powders. Non freeze-driedM. oleifera was prepared into different packaging namely open container, closed container and vacuum packing, whilst, freeze-driedM. oleifera was stored in closed container and vacuum packing. Each of the packaging was stored at room temperature (30 to 32°C) and refrigerator (4°C). The turbidity removal efficiencies of storedM. oleifera were examined using jar test at monthly interval for 12 months. The results indicated that non freeze-driedM. oleifera kept in the refrigerator (4°C) would preserve its coagulation efficiency. In addition, closed container and vacuum packing were found to be more appropriate for the preservation of non freeze-driedM. oleifera, compared to open container. Freeze-driedM. oleifera retained its high coagulation efficiency regardless the storage temperature and packaging method for up to 11 months. Besides, higher increment in zeta potential values for water coagulated with freeze-driedM. oleifera indicated the higher frequency of charge neutralization and better coagulation efficiency of freeze-driedM. oleifera, compared to non freeze-dried seeds. As a coagulant,M. oleifera did not affect the pH of the water after treatment.

Nitrate removal in a packed bed reactor using volatile fatty acids from anaerobic acidogenesis of food wastes

Abstract: A packed bed reactor (PBR) was fed with nitrate containing synthetic wastewater or effluent from a sequencing batch reactor used for nitrification. The C source introduced into the PBR consisted of volatile fatty acids (VFAs) produced from anaerobic acidogenesis of food wastes. When nitrate loading rates ranged from 0.50 to 1.01 kg N/m3·d, the PBR exhibited 100∼98.8% NO3−-N removal efficiencies and nitrite concentrations in the effluent ranged from 0 to 0.6 NO2−-N mg/L. When the PBR was further investigated to determine nitrate removal activity along the bed height using a nitrate loading rate less than 1.01 kg N/m3·d, 100% nitrate removal efficiency was observed. Approximately 83.2% nitrate removal efficiency was observed in the lower 50% of the packed-bed height. When reactor performance at a C/N ratio of 4 and a C/N ratio of 5 was compared, the PBR showed better removal efficiency (96.5%) of nitrate and less nitrite concentration in the effluent at the C/N ratio of 5. VFAs were found to be a good alternative to methanol as a carbon source for denitrification of a municipal wastewater containing 40 mg-N/L.

Antioxidant properties of Erigeron annuus extract and its three phenolic constituents

Abstract: The antioxidant activity of the extract ofErigeron annuus was assessed by means of two differentin vitro tests: bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH test) and the scavenging of authentic peroxynitrite in company with peroxynitrite generation from 3-morpholinosydnonimine (SIN-1). In both tests, the 85% aq. MeOH andn-BuOH soluble fractions of the crude extract showed a significant scavenging effect on peroxynitrite and DPPH radical in comparison to L-ascorbic acid. And bioassay-guided fractionation of then-BuOH soluble fraction led to the isolation of three compounds: Apigenin (1), quercetin-3-O-glucoside (2), and caffeic acid (3). The structures of the isolated compounds were elucidated on the basis of their spectroscopic data and their antioxidant activities were measured by determining their capacity to scavenge peroxynitrite and the DPPH radical.

Lactic Acid Fermentation of Chinese Yam (Dioscorea batatas Decne) Flour and Its Pharmacological Effect on Gastrointestinal Function in Rat Model

Abstract: To develop a health-aid preparation of Chinese yam (Dioscorea batatas Decne), lactic acid fermentation was attempted using a mixed starter comprising ofLactobacillus acidophilus, Streptococcus thermophilus, andBifidobacterium bifidus. The anaerobic fermentation of a 5% Chinese yam flour suspension gave a uniform suspension of pH 4.35, containing 7.76×106 CFU/mL lactic acid bacteria (LAB), and which was found to be acceptable to the panel from a sensory assessment. During the administration of the lactic acid fermented (LAF) Chinese yam to Sprague Dawley rats for 6 weeks, a smaller body weight gain, but greater excretion of feces were observed, implying the creation of a healthy gastrointestine on the administration of LAF Chinese yam, which was also confirmed by the gastrointestinal motility of the feed in rats fed on LAF Chinese yam. The constipation induced by loperamide was further suppressed in a rat group fed on a LAF Chinese yam diet, which was qualified from healthy gastrointestinal flora established by LAB. A serochemical analysis revealed a slight improvement in the blood glucose, neutral lipid and total cholesterol concentrations on administration of LAF Chinese yam, suggesting LAF Chinese yam could be served as a healthy-aid preparation, even for hyperglycemia or hyperlipidemia patients.

Anaerobic Ammonium Oxidation Process in an Upflow Anaerobic Sludge Blanket Reactor with Granular Sludge Selected from an Anaerobic Digestor

Abstract: The purpose of this work was to evaluate the development of the anammox process by the use of granular sludge selected from a digestion reactor as a potential seed source in a lab-scale UASB (upflow anaerobic sludge blanket) reactor system. The reactor was operated for approximately 11 months and was fed by synthetic wastewater. After 200 days of feeding with NH4 + and NO2 − as the main substrates, the biomass showed steady signs of ammonium consumption, resulting in over 60% of ammonium nitrogen removal. This report aims to present the results and to more closely examine what occurs after the onset of anammox activity, while the previous work described the start-up experiment and the presence of anammox bacteria in the enriched community using the fluorescencein situ hybridization (FISH) technique. By the last month of operation, the consumed NO2 −N/NH4 +-N ratio in the UASB reactor was close to 1.32, the stoichiometric ratio of the anammox reaction. The obtained results from the influentshutdown test suggested that nitrite concentration would be one key parameter that promotes the anammox reaction during the start-up enrichment of anammox bacteria from granular sludge. During the study period, the sludge color gradually changed from black to red-brownish.

Recovery of lactic acid from fermentation broth by the two-stage process of nanofiltration and water-splitting electrodialysis

Abstract: A two-stage process of nanofiltration and water-splitting electrodialysis was investigated for lactic acid recovery from fermentation broth. In this process, sodium lactate is isolated from fermentation broth in the first stage of nanofiltration by using an NTR-729HF membrane, and then is converted to lactic acid in the second stage by water-splitting electrodialysis. To determine the optimal operating conditions for nanofiltration, the effects of pressure, lactate concentration, pH and known added impurities were studied. Lactate rejection was less than 5%, magnesium rejection approximated 45%, and calcium rejection was at 40%. In subsequent water-splitting electrodialysis, both the sodium lactate conversion to lactic acid and sodium hydroxide recovery, were about 95%, with a power requirement of 0.9∼1.0 kWh per kg of lactate.

Self-organized pullulan/deoxycholic acid nanogels: Physicochemical characterization and anti-cancer drug-releasing behavior

Abstract: The objective of this study was to develop new self-organized nanogels as a means of drug delivery in patients with cancer. Pullulan (PUL) and deoxycholic acid (DOCA) were conjugated through an ester linkage between the hydroxyl group in PUL and the carboxyl group in DOCA. Three types of PUL/DOCA conjugates were obtained, differing in the number of DOCA substitutions (DS; 5, 8, or 11) per 100 PUL anhydroglucose units. The physicochemical properties of the resulting nanogels were characterized by dynamic light scattering, transmission electron microscopy, and fluorescence spectroscopy. The mean diameter of DS 11 was the smallest (approx. 100 nm), and the size distribution was unimodal. To determine the organizing behavior of these conjugates, we calculated their critical aggregation concentrations (CACs) in a 0.01-M phosphate buffered saline solution. They were 10.5×10−4 mg/mL, 7.2×10−4 mg/mL, and 5.6×10−4 mg/mL for DS 5, 8, and 11, respectively. This indicates that DOCA can serve as a hydrophobic moiety to create self-organized nanogels. To monitor the drug-releasing behavior of these nanogels, we loaded doxorubicin (DOX) onto the conjugates. The DOX-loading efficiency increased with the degree of DOCA substitution. The release rates of DOX from PUL/DOCA nanogels varied inversely with the DS. We concluded that the PUL/DOCA nanogel has some potential for use as an anticancer drug carrier because of its low CAC and satisfactory drug-loading capacity.

Molecularly imprinted monolithic stationary phases for liquid chromatographic separation of tryptophan andN-CBZ-phenylalanine enantiomers

Abstract: Monolithic molecularly imprinted columns were designed and prepared by anin-situ thermal-initiated copolymerization technique for rapid separation of tryptophan andN-CBZ-phenylalanine enantiomers. The influence of polymerization conditions and separation conditions on the specific molecular recognition ability for enantiomers and diastereomers was investigated. The specious molecular recognition was found to be dependent on the stereo structures and the arrangement of functional groups of the imprinted molecule and the cavities in the molecularly imprinted polymer (MIP). Moreover, hydrogen bonding interactions and hydrophobic interactions played an important role in the retention and separation. Compared to conventional MIP preparation procedures, the present method is very simple, and its macroporous structure has excellent separation properties.

The growth and EPA synthesis ofShewanella oneidensis MR-1 and expectation of EPA biosynthetic pathway

Abstract: Shewanella oneidensis MR-1 has the ability to inhale certain metals and chemical compounds and exhale these materials in an altered state; as a result, this microorganism has been widely applied in bioremediation protocols. However, the relevant characteristics of cell growth and biosynthesis of PuFAs have yet to be thoroughly investigated. Therefore, in this study, we have attempted to characterize the growth and fatty acid profiles ofS. oneidensis MR-1 under a variety of temperature conditions. The fastest growth ofS. oneidensis MR-1 was observed at 30°C, with a specific growth rate and doubling time of 0.6885 h−1 and 1.007 h. The maximum cell mass of this microorganism was elicited at a temperature of 4°C. The eicosapentaenoic acid (EPA) synthesis ofS. oneidensis MR-1 was evaluated under these different culture temperatures.S. oneidensis MR-1 was found not to synthesize EPA at temperatures in excess of 30°C, but was shown to synthesize EPA at temperatures below 30°C. The EPA content was found to increase with decreases in temperature. We then evaluated the EPA biosynthetic pathway, using a phylogenetic tree predicted on 16s rRNA sequences, and the homology of ORFs betweenS. oneidensis MR-1 andShewanella putrefaciens SCRC-2738, which is known to harbor a polyketide synthase (PKS)-like module. The phylogenetic tree revealed that MR-1 was very closely related to bothMoritella sp., which is known to synthesize DHA via a PKS-like pathway, andS. putrefaciens, which has been reported to synthesize EPA via an identical pathway. The homology between the PKS-like module ofS. putrefaciens SCRC-2738 and the entire genome ofS. oneidensis MR-1 was also analyzed, in order to mine the genes associated with the PKS-like pathway inS. oneidensis MR-1. A putative PKS-like module for EPA biosynthesis was verified by this analysis, and was also corroborated by the experimental finding thatS. oneidensis MR-1 was able to synthesize EPA without the expression of dihomo-γ-linoleic acid (DGLA) and arachidonic acid (AA) formed during EPA synthesis via the FAS pathway.

Kinetics and Modelling of Cell Growth and Substrate Uptake in Centella asiatica Cell Culture

Abstract: In this study, we have conducted kinetics and modelling studies ofCentella asiatica cell growth and substrate uptake, in an attempt to evaluate cell growth for a better understanding and control of the process. In our bioreactor cultivation experiment, we observed a growth rate of 0.18/day, a value only 20% higher than was seen in the shake flask cultivation trial. However, the observed maximum cell dry weight in the shake flask, 10.5 g/L, was 14% higher than was achieved in the bioreactor. Ninety seven percentage confidence was achieved via the fitting of three unstructured growth models; the Monod, Logistic, and Gompertz equations, to the cell growth data. The Monod equation adequately described cell growth in both cultures. The specific growth rate, however, was not effectively predicted with the Logistic and Gompertz equations, which resulted in deviations of up to 73 and 393%, respectively. These deviations in the Logistic and Gompertz models may be attributable to the fact that these models were developed for substrate-independent growth and fungi growth, respectively.

Enhanced production of laccase fromTrametes sp. by combination of various inducers

Abstract: In this study, we have attempted to determine the optimum concentration of inducers responsible for efficient laccase production by the white-rot fungus,Trametes sp. Variations in laccase activity were investigated with changing concentrations of 2,5-xylidine, syringaldazine, ABTS, and guaiacol. Enhancement of peak laccase activity was achieved via the combination of 2,5-xylidine with ABTS, syringaldazine, or guaiacol, resulting in increases of up to 359, 313, and 340%, respectively, as compared to control values. Among the tested inducers, the addition of 0.1 mM of ABTS coupled with 1.0 mM of 2,5-xylidine in the medium after 24 h of cultivation proved optimal with regard to laccase enzyme production.

The preparation of D-phenylalanine imprinted microbeads by a novel method of modified suspension polymerization

Abstract: Molecularly imprinted polymeric microbeads (MIPMs) were prepared by the suspension and modified suspension polymerization methods using D-phenylalanine as the template, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linker, toluene as the porogen, polyvinyl alcohol as the stabilizer, and sodium dodecyl sulfate as the surfactant. The addition of a surfactant to the conventional suspension polymerization mixture decreased the mean particle size of the MIPMs and increased the adsorption selectivity. For the modified suspension polymerization method, the mean particle size of the MIPMs was smaller than the particle size of MIPMs prepared via conventional suspension polymerization. Moreover, the adsorption selectivity improved considerably compared to the adsorption selectivities of MIPs reported previously.

Specific detection of DNA using quantum dots and magnetic beads for large volume samples

Abstract: Here we present a sensitive DNA detection protocol using quantum dots (QDs) and magnetic beads (MBs) for large volume samples In this study, QDs, conjugated with streptavidin, were used to produce fluorescent signals while magnetic beads (MBs) were used to isolate and concentrate the signals The presence of target DNAs leads to the sandwich hybridization between the functionalized QDs, the target DNAs and the MBs In fact, the QDs-MBs complex, which is bound using the target DNA, can be isolated and then concentrated The binding of the QDs to the surface of the MBs was confirmed by confocal microscopy and Cd elemental analysis It was found that the fluorescent intensity was proportional to concentration of the target DNA, while the presence of non-complementary DNA produced no significant fluorescent signal In addition, the presence of low copies of target DNAs such as 05 pM in large volume samples up to 40 mL was successfully detected by using a magnet-assisted concentration protocol which consequently results in the enhancement of the sensitivity more than 100-fold

Effects of morphology and rheology on neofructosyltransferase production byPenicillium citrinum

Abstract: In this study, we investigated the relationship between the morphology and the rheological properties ofPenicillium citrinum to improve the production of neo-fructosyltransferase (neo-FTase). In a 2.5 L bioreactor culture ofP. citrinum, it was observed that agitation speed and aeration rate had significant effects on the production of neo-FTase and that maximum cell mass and neo-FTase production obtained at 500 rpm and 1.5 vvm were 8.14 g/L and 53.2×10−3 U/mL, respectively. Cell mass and neo-FTase production increased to 91.53 and 25.17%, respectively. In the morphology and rheology studies,P. citrinum showed a typical pellet morphology that was explained by a shaving mechanism; this phenomenon was significantly affected by carbon sources. The rheology of neo-FTase fermentation byP. citrinum was dependent on cell growth and fungal morphology.

Screening of Korean marine plants for their inhibitory effect on histamine release from RPMCin vitro

Abstract: Allergy, meaning ‘heightened reactivity’ of a host on being exposed to an antigen, is an immediate reaction which included anaphylaxis following contact with an antigen. An anaphylatic reaction is caused by the release of pharmacological mediators, like histamine, from mast cells. The potential anti-allergic activities of 27 seaweed and 19 salt marsh extracts collected from the coast of Korea were tested against the inhibition of histamine release in rat peritoneal mast cells (RPMCs). Among them, three salt marsh plants (Persicaria lapathifolia, Ixeris tamagawaensis, andSalsola komarovir) significantly showed more than 75% of inhibition of the histamine release at a concentration of 100 μg/mL, and also three salt marsh (Messerschmidia sibirica, Rosa rugosa, andPortulaca oleraceae) and three seaweed (Colpomenia bullosa, Derbesia marina, andSargassum thunbergii) extracts exhibited moderately inhibition effects when compared to the control.

Enzymatic release of ferulic acid from Ipomoea batatas L. (sweet potato) stem

Abstract: Ferulic acid is a phenolic compound that serves as a major biosynthetic precursor of vanillin in higher plants. We investigated the ability of the 3 commercial enzymes—Ultraflo L, Viscozyme L, and α-Amylase—to induce the release ferulic acid from theIpomoea batatas L. (sweet potato) stem. The rate of release for ferulic acid was optimal when Ultraflo L (1.0%) was used compared with the other enzymes, whereas Viscozyme L was most effective for the release of vanillic acid and vanillin. Thus, these enzymes may be useful for the large-scale production of ferulic acid and other phenolic compounds from sweet potato stem.

High-yield Production of Functional Human Lactoferrin in Transgenic Cell Cultures of Siberian Ginseng (Acanthopanax senticosus)

Abstract: Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducibleSWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.

Removal of Aqueous Pentachlorophenol by Horseradish Peroxidase in the Presence of Surfactants

Abstract: An important issue in the oxidation of pentachlorophenol (PCP) by the enzyme horseradish peroxidase (HRP) is enzyme inactivation during the reaction. This study was initiated to investigate the ability of two nonionic surfactants (Tween 20 and Tween 80) to mitigate HRP inactivation. The surfactants were tested at concentrations below and above their critical micelle concentrations (CMCs). Enhancement of PCP oxidation was observed at sub-CMCs, indicating effective protection of HRP by the two surfactants. Maximum levels of PCP removal were observed when the concentrations of Tween 20 and Tween 80 were 40 and 50% of the CMCs, respectively. At supra-CMCs, both surfactants caused a noticeable reduction in the extent of PCP removal.

Effects of hydrocarbon additions on gas-liquid mass transfer coefficients in biphasic bioreactors

Abstract: The effects of aliphatic hydrocarbons (n-hexadecane andn-dodecane) on the volumetric oxygen mass transfer coefficient (k L a) were studied in flat alveolar airlift reactor and continuous stirred tank reactors (CSTRs). In the flat alveolar airlift reactor, high aeration rates (>2 vvm) were required in order to obtain efficient organic-aqueous phase dispersion and reliablek L a measurements. Addition of 1% (v/v)n-hexadecane orn-dodecane increased thek l a 1.55-and 1.33-fold, respectively, compared to the control (superficial velocity: 25.8×10−3 m/s, sparger orifice diameter: 0.5 mm). Analysis of the gas-liquid interfacial areaa and the liquid film mass transfer coefficientk L suggests that the observedk L a increase was a function of the media's liquid film mass transfer. Addition of 1% (v/v)n-hexadecane orn-dodecane to analogous setups using CSTRs led to ak L a increase by a factor of 1.68 and 1.36, respectively (superficial velocity: 2.1×10−3 m/s, stirring rate: 250 rpm). These results propose that low-concentration addition of oxygen-vectors to aerobic microbial cultures has additional benefit relative to incubation in purely aqueous media.

Characterization of a newly isolatedcis-1,2-dichloroethylene and aliphatic compound-degrading bacterium,Clostridium sp. strain KYT-1

Abstract: Acis-1,2-dichloroethylene (cis-DCE)-degrading anaerobic bacterium,Clostridium sp. strain KYT-1, was isolated from a sediment sample collected from a landfill site in Nanji-do, Seoul, Korea. The KYT-1 strain is a gram-positive, endospore-forming, motile, rod-shaped anaerobic bacterium, of approximately 2.5∼3.0 μm in length. The degradation ofcis-DCE is closely related with the growth of the KYT-1 strain, and it was stopped when the growth of the KYT-1 strain became constant. Although the pathway ofcis-DCE degradation by strain KYT-1 remains to be further elucidated, no accumulation of the harmful intermediate, vinyl chloride (VC), was observed during anaerobiccis-DCE degradation. Strain KYT-1 proved able to degrade a variety of volatile organic compounds, including VC, isomers of DCE (1,1-dichloroethylene,trans-1,2-dichloroethylene, andcis-DCE), trichloroethylene, tetrachloroethylene, 1,2-dichloroethane, 1,1,1-trichloroethane, and 1,1,2-trichloroethane. Strain KYT-1 degradedcis-DCE at a range of temperatures from 15 to 37°C, with an optimum at 30°C, and at a pH range of 5.5 to 8.5, with an optimum at 7.0.

Antitumor activity of cell suspension culture of green tea seed (Camellia sinensis L.)

Abstract: The objective of this study was to investigate the antitumor activity of suspension cultures of tea callus cells grown in the presence of different concentrations of the growth regulator 2,4-dichlorophenoxy acetic acid (2,4-D) with or without light irradiation. The methanol and ethanol extracts of precipitated cells (MEP, EEP) exhibited stronger inhibitory effects on the growth of tumor cell lines than the water extract of precipitated cells (WEP) or the supernatant. Compared to culture under dark conditions, exposure to light irradiation led to significantly higher antitumor activity. The MEP from light irradiated cells at 250 μg/mL with 2.0 mg/L 2,4-D displayed more than 64% growth inhibition of HEP-2 cells, whereas normal cells showed less than 25% growth inhibition. The some fractions of MEP obtained from Diaion HP-20 column chromatography displayed the majority of inhibitory activity against the HEP-2 cell line. These results show that 2,4-D, and light stimulated the synthesis of antitumor compounds.