Earticle

Aptamers are nucleic acid species that have been engineered through repeated rounds, SELEX (systematic evolution of ligands by exponential enrichment) to bind to various molecular targets. They offer the utility for biotechnological and therapeutic applications as they have molecular recognition properties that rival that of the commonly used biomolecule, antibodies. Carcinoembyonic antigen(CEA) is a glycoprotein involved in cell adhesion. It was found that serum from individuals with colorectal, gastric, pancreatic, lung and breast carcinomas had higher levels of CEA than healthy individuals. CEA measurement is mainly used as a tumor marker to identify recurrences after surgical resection. First, CEA has bond on the magnetic bead through chemical bonding and it was confirmed by CEA specific antibody(ELISA). Second, We synthesized a labeled(fluorescence) single strand DNA(ssDNA) for CEA, using asymmetric PCR with a 5’-labeled primer. Finally, through the repeated rounds of SELEX, specific ssDNA was recovered and verified by its binding affinity.

We investigated the effects of temperature and the addition of detergents on the asymmetric hydrolysis reaction catalyzed by the recombinant Escherichia coli containing the epoxide hydrolase (EH) gene of Rhodotorula gluticis. The hydrolysis rate and enantioselectivity were increased in the presence of various detergents such as Tween 20, especially at low reaction temperature. The reaction conditions were optimized, and chiral 98 %ee (S)-styrene oxide could be prepared by using batch asymmetric hydrolysis reaction. Acknowledgement: This work was supported by the Marine and Extreme Genome Research Center Program, Ministry of Land, Transportation and Maritime Affairs, Republic of Korea.

Lipase is a well-known enzyme and used as a biocatalyst for chemical and food processes and bio-diesel production. The calB gene coding for lipase B was cloned from the Candida antarctica ATCC32657 genomic DNA. Recombinant CalB fused with the 10 arginine (R10) or lysine (K10) tag at its C-terminal was expressed in Escherichia coli . SDS PAGE-analysis of CalB expression indicated that the band densities of soluble lipase B fused with R10 (CalB-R10) and K10 (CalB-K10) were 2.2 and 1.8 times higher than that of CalB without any tags, respectively. Activity assay of the crude protein extract using para-nitrophenyl palmitate as a substrate showed that CalB-R10 and CalB-K10 possessed 31.7±0.28 U/mg and 26.2±0.37 U/mg of specific activities, respectively, which were 1.5-1.9 times higher than that of CalB (17.1±0.6 U/mg). Incubation of the soluble CalB-R10 at pH9.0 and CalB-K10 at pH10.0 for 7 days resulted in 49% and 45% decreases in specific activity, respectively, corresponding to a nearly 2-fold increase compared with specific CalB activity.

Cytochrome P450s act on the inactive carbon-hydrogen bonds of alkanes, fatty acids, terpenes, and steroids; and some exhibit high regio and stereoselective monooxygenation activity. And cytochrome P450s are expected to be potential catalysts for fine chemical synthesis. Regiospecific hydroxylation of isoflavone using cells of Bacteria, actinomyce was examined. Recent researches reported hydroxylated isoflavones have an effect on the anticancer, antitumor, and antioxidant. Daidzein, the major soy isoflavone was finally hydroxylated by whole cell reaction with Streptomyces avermitilis MA4680 and Nocardia farcinica IFM10152. The major hydroxylated products of daidzein were 7,3’,4’-trihydroxyisoflavone, 6,7,4’- trihydroxyisoflavone and 7,8,4’-trihydroxyisoflavone which were mono-hydroxylated at ortho position of hydroxy group of daidzein. Also we identified P450 monooxygenases involved in monohydroxylation of daidzein and expressed in E. coli. P450s need redox partner for hydroxylation to transfer electrons to the heme domain of P450 and activate the hydroxylation step of the substrate-heme domain complex. We constructed P450s and redox partners which matched best in E. coli . The hydroxylated products were determined using high performance liquid chromatograph (HPLC), gas chrmomatograph / mass spectrometry (GC/MS) Acknowledgement : This research was supported by a grant from the National Research Laboratory Program of the Korea Science and Engineering Foundation.(Grant No. ROA-2007-000-10007-0)

The production of biodiesel by enzymatic transesterification has attracted much attention for high purity product and enables easy separation. Candida antarctica lipase B is one of the most important enzyme in the biodiesel reaction. In reaction, CALB shows low organic solvent stability while having relation with methanol. In this study, we concerned about enhancing organic solvent stability in hydrophilic solvent. In water-miscible solvents, water molecule seems to play an important role in biological structure and function. Water molecules in the enzyme are stripped away or replaced with hydrophilic solvent molecules and this causes deformation and denaturation of the enzyme. From the directed evolution studies in hydrophilic solvents, introduction of polar or charged amino acids increased hydrogen bonds and increased enzyme stability. Increase of hydrogen bondinteraction can make water molecule less strips off and more stable in hydrophilic organic solvents. Hydrogen bond interaction can be increased by shorten the distance of amino acids and water molecule and also increase electrostatic interaction. ASP and ASN was replaced to GLU and GLN of CALB for in silico mutagenesis. Mutants were analyzed by hydrogen bonding network and solvent accessible surface area.

Candida antarctica lipase B(CALB) is an important catalyst in bio-organic synthesis. CALB was successfully expressed in the yeast P ichia pastoris and Saccaromyces cerevisiea. However, the use of E.coli for the production of CALB has faced many practical and biological problems such as insoluble aggregates and proteolytic degradation. In this research, for the enhancement expression of CALB in E.coli , CALB was expressed in periplasm using PelB signal sequence. And the research of chaperon-CALB fusion was performed to improve the expression level and solubility of CALB in E. coli .

We have investigated a number of epoxide hydrolase (EH)-catalyzed enantioconvergent biohydrolysis for the production of chiral diols with a yield greater than 50%. All these enantioconvergent biohydrolysis have been carried out on the basis of stereochemical flexibilities of Caulobacter crescentus EH (CcEH). Enantioconvergent biohydrolysis using single CcEH or two EHs in combination with CcEH have been optimized and compared for their feasibility for the production of chiral (R)-phenyl-1,2-ethanediol from racemic styrene oxide as a model substrate. Acknowledgment: This work was partially supported by the Marine and Extreme Genome Research Center Program, Ministry of Marine Affairs and Fisheries, Korea.

Protease digestion efficacy is a critial parameter in an initial protein identification process by mass spectrometry (MS). However, some of proteins are resistant to proteolysis because of their compact conformation from extensive β-sheet structure [1]. Hepatitis B surface antigen (HBsAg) particle is a lipid-associated large protein complex (> 2,200 kDa) [2]. HBsAg was digested by two kinds of protease (trypsin and pepsin) and analyzed by RP-HPLC and MS to investigate the effective digestion conditions. HBsAg was denatured with thermal and chemical treatment before tryptic digestion. The sequence coverage of HBsAg (4%) was not improved after thermal treatment (80℃, 3 min). After 1% sodium deoxycholate (SDC) treatment, the sequence coverage of HBsAg was improved upto 13%, but not enough to be identified. Even after 1% SDS and 1% merchaptoethanol treatment at 60℃ for 1 min to fully dissociate the particle structure to HBsAg monomers, no peak was eluted by RP-HPLC. This was because tryptic cleavage sites of HBsAg were hidden by the strong compact structure. However, peptide fragments of HBsAg treated with pepsin after 1% SDS and 1% merchaptoethanol treatment at 60℃ for 1 min were eluted by RP-HPLC. Therefore, pepsin appeared more suitable than trypsin as a HBsAg-digestion protease. In this presentation, the data of RP-HPLC chromatograms and MS spectra are presented for each cleavage condition.

A novel GDSLesterase from Sinorhizobium meliloti 1021 (EstSM), which has conserved amino acid sequences of Gly8, Asn9, Ser10, and Leu11 in N-terminal region, is characterized. Molecular weight of EstSM is 23kDa, and its pI is 4.89, respectively. EstSM has ~40% sequence identities with Mycobacterium smegmatis arylesterase(PDB ID: 2Q0Q) with putative catalytic sites of Ser10, Asp187, and His190. Using pQE30 vector, EstSM was expressed in Eschrichia coli BL21 codon plus, and purified by His-tag affinity column. Activities of EstSM are investigated using various substrates such as p-nitrophenyl acetate and (R,S)-methyl-β-hydroxyisobutyrate. Biochemical and structural characterization of EstSM will provide the specificity regulation and activation mechanism of GDSL family in more detail. In addition, crystal studies for three dimensional structures are in progress.

We observed that an inclusion body (IB) of recombinant β-galactosidase that was produced by the araBAD promoter system in Escherichia coli (E. coli) showed enzyme activity. In order to improve its activity, the lowering of the transcription rate of the β-galactosidase structural gene was attempted through competition between an inducer (L-arabinose) and an inducer analog (D-fucose). In the deep-well microtiter plate culture and lab-scale fermentor culture, it was demonstrated that the addition of D-fucose caused an improvement in specific β-galactosidase production, although β-galactosidase was produced as an IB. In particular, the addition of D-fucose after induction led to an increase in the specific activity of β-galactosidase IB. Finally, we confirmed that the addition of D-fucose after induction caused changes in the structure of β-galactosidase IB with higher enzyme activity. Based on these results, we expect that an improved enzyme IB will be used as a biocatalyst of the enzyme bioprocess because an enzyme IB can be purified easily and has physical durability.

The functional Lipase B from Candida antarctica(CalB) was expressed in a cell-free protein synthesis system derived from Escherichia coli . While most of the cell-free synthesized CalB was insoluble under conventional reaction conditions, the combined use of molecular chaperones led to the soluble expression of CalB. Inaddition, the functional enzyme was generated by applying the optimal redox potential. When examined using pNPP as a substrate, the specific activity of the cell-free synthesized CalB was higher than that of the reference protein produced in Pichia pastoris. These results highlight the potential of cell-free protein synthesis technology as a powerful platform for the rapid expression, screening and analysis of industrially important enzymes.

A recombinant Escherichia coli harboring heterologous styrene monooxygenase (SMO) genes under the control of T7 promoter produced protein inclusion bodies extensively and exhibited a low SMO activity. To express SMO in soluble form and increase the whole-cell SMO activity, various chaperones including DnaK-DnaJ-GrpE, GroES-GroEL and trigger factor were co-expressed in the recombinant E. coli individually or in combinations. Co-expression of chaperones improved the SMO activity greatly, especially with combined expression of DnaK-DnaJ-GrpE and GroESGroEL as 180 U/g cell, which was almost 2-fold higher than the one without chaperons overexpression. According to SDS-PAGE analysis, the production of SMO proteins in the soluble fraction of cell lysate increased by chaperones co-expression. The high SMO activity was maintained during water-organic two-phase experiment for 12 h, which yielded 170 mM enantiopure (S)-styrene oxide in organic phase. This study illustrates that chaperones co-expression can be an efficient strategy for the development of an active whole-cell biocatalyst.

alyVI is a gene which encodesan alginate lyase of marine bacterium Vibrio sp. QY101 (338aa). A nine-amino-acid consensus region (YXRESLREM), which was only found in poly-G lyases, was also observed in the amino-terminal region of alyVI. However, alyVI could degrade both M block and G block. A1-II’ is a close homolog with a reported structure (2CWS). A 3D structure of this enzyme was modeled based on this template by the comparative modeling technique. As in the template alginate, tyrosine seems critical for enzymatic activity. Since it shows enzymatic activity for poly-M alginate, docking a tri-mannuronate as well as a tri-guluronate into the enzyme was performed. The ligand was nicely posed inside of the each binding site with a strikingly similar way. In accordance with previous report, OH-group of a tyrosine is close to -hydrogen, suitable to initiate -elimination reaction. By considering both ligands, it can be designed to change stereoselectivity of this enzyme to either M-specific or G-specific in a rational way.

Clinical studies have demonstrated that a prostaglandin E2 (PGE2) analogue had the property of increasing hair density and controlling hair loss in humans. However, prostaglandins are molecules which have a very short biological half-life period and which act in an autocrine or paracrine manner, reflecting a labile character due to a very active local metabolism of the prostaglandins. It therefore appears to be important, in order to maintain and/or increase hair density in humans, to preserve the endogenous reserves of PGE2 of the different compartment of the hair follicle or of its close cutaneous environment. NAD+-linked 15-hydroxyprostaglandin dehydrogenase (15-PGDH) catalyzes the oxidation of 15(S)-hydroxyl group of prostaglandins and lipoxins resulting in the formation of 15-keto metabolites which exhibit greatly reduced biological activities. Therefore, this enzyme has been considered the key enzyme responsible for the inactivation of prostaglandins and lipoxins. Inhibitors of this enzyme may be of value on the hair density and/or growth. Previously, ciglitazone, an anti-diabetic thiazolidinedione, was found to be a potent inhibitor of 15-PGDH. Structure-activity analysis of available thiazolidinediones indicated that hydrophobicity of the moiety linking to phenyl ring through ether linkage as well as benzylidene configuration play important roles in inhibitory potency. Furthermore, N-methylation of 2,4-thiazolidinedione abolished the inhibitory activity. A series of benzylidene thiazolidinediones with varied ring structure and methylene bridge to phenyl ring through ether linkage were synthesized and assayed for inhibitory activity. It was found that compound CT-8 (5-[4-(cyclohexylethoxy)benzylidene]-2,4-thiazolidinedione) was the most potent inhibitor effective at nM range.

Rhynchosia nulubilis has been used as supplements of estrogen for preventing postmenopausal osteoporosis and as food in oriental medicine. Contents of free amino acid of germinated R. nulubilis seeds treated by low molecular weight soluble chitosan(LMSC) were investigated. R. nulubilis seeds were germinated to 2 mm-length root at 20℃ after presoaking in 0.05% LMSC(1, 3, 5, 10 kDa) solution for 4hr. Total free amino acid content of 5 kDa chitosan-treated group was higher than those of control and the other group, also total essential amino acid content in all chitosan-treated group was higher than those of control. Arginine(Arg), tyrosine(Tyr), valine(Val), methionine(Met), tryptophan(Trp) contents in treated group were higher than those of control. Especially, 5 kDa chitosan of various molecular weight of chitosan was highest content which showed in Arg(331 mg%), Tyr(19 mg%), Val(36 mg%), Met(34 mg%), Trp(53 mg%). In the Met content, chitosan-treated group was four time than control(8 mg%), 33~34 mg% in chitosan-treated group.

Anthriscus sylvestris is a perennial herb which belongs to the Umbelliferae family and its dried root has been used as a herbal medicine in Korea, Chian. The roots are used in therapy for digestion promotion, nutrition strengthening, the pollakiuria of old age, the discharge of phlegm, the removal of fever, and toothache. This experiment was conducted to investigate cytotoxicity of the ethanol extracts of stem, leaves, root, and whole plant from Anthriscus sylvestris against a human gastric carcinoma cell (SNU-601 cell line) and a human pulmonary carcinoma cell (Calu-6 cell line). The results showed that the cytotoxicity of these extracts were leaves, stem, whole plant, and root against two cell lines in sequence. In particular, leaves extract at the concentration of 800 ㎍/㎖ was inhibited above ninety percent on the growth of two cancer cell lines and at the concentration of 400 ㎍/㎖ was inhibited about fiftytwo percent and seventyseven percent against SNU-601, Calu-6 cell growth, respectively. Stem extract at 800 ㎍/㎖ was also inhibited above fifty percent against two cancer cell lines. This result represents that Anthriscus sylvestris would be a potential antitumor resource.

Ferritin is a major iron storage protein that is widely distributed in animal and plant tissues, and consists of 24 subunits to form a hollow shell with an outer diameter of 12 nm and an inner diameter of 8 nm. The iron core naturally existing inside the ferritin was found to be readily extracted and replaced with various materials such as metals, metal oxides, and semiconductors. Thus, ferritins can act as a nano-scale template, a biocompatible cage, and a separator between the nanoparticles. In the present study, ferritins were chemically reconstituted with cobalt and manganese oxyhydroxides. The formation of Co(O)OH within the apoferritin interior was conducted in MOPS and HEPES buffers at pH 8.3-8.5 using hydrogen peroxide as the oxidant and Mn(O)OH-reconstituted ferritin was prepared in AMPSO buffer at pH 8.9 using air as the oxidant. The synthesis of Co(O)OH and Mn(O)OH in the cavity of apoferritin was monitored by UV-visible spectrophotometry for ranging from 300 to 800 nm. It is expected that ferritins reconstituted with cobalt and manganese oxyhydroxides will have a variety of applications due to their chemical reactivity and electrochemical activity.

In vitro biological activities such as antioxidant activity, whitening activity, anti-hangover activity and anticancer activity were evaluated. The antioxidant activities of astaxanthin and H . pluvialis extract were significantly higher than that of α-tocopherol when the antioxidant activities were determined as the nitrate scavenging activity, hydroxyl radical scavenging activity and DPPH radical scavenging activity. The whitening effect of H . pluvialis extract was about two times as kojic acid. H . pluvialis extract indicated an anticancer activity on a lung cancer cell line and a cervical cancer cell line. Astaxanthin showed anti-hangover effect of 1.5 times as jiguja extract. The anti-hangover effect of the inclusion complex (As-β-CDIC) was about 1.2 times of jiguja extract. And, the inclusion complex of H . pluvialis (H.p.-CycloBIC) showed almost the same whitening effect as kojic acid.

In previous reports, anti-oxidant effect of deproteinised silkworm hemolymph (DSH) was confirmed by its ability to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals in in vitro experiments. In vivo level stamina enhancement activity of DSH, extracted from 5th instar larvae of Bombyx mori, was investigated on Sprague Dawley rats via a weight-loaded forced-swimming test using Pb as the weight. Oral administration of DSH at a concentration of 1.0 ml/kg body weight to rats was given daily for 7 and 14 days using sonde as the feeding apparatus. The rats had undergone 5 min/day swimming to accustom them to swimming. To limit the circadian variations, fasting, feeding and swimming times were consistent. After 7 and 14 days, final swimming times were assessed as the point of exhaustion by their inability to rise back to the surface for 7 seconds. Comparative results showed that 7 day and 14 day DSH administered group had a remarkable 34.7% and 4.2% increase in the swimming time than the 3 step filtered water group (control), respectively. From our findings, we have verified that stamina increases with respect to increase in the period of DSH intake which demonstrates the intake period dependency of DSH on stamina. The findings of this study indicate that DSH dosage of 1.0 ml/kg body weight has a significant stamina enhancing effect and opens the possibility as a potential energy beverage or usage as a therapeutic medicine.

Chitooligosaccharides (COSs) are derivative of chitosan and it can be obtained by either enzymatic or chemical hydrolysis of chitosan. COSs are not only water-soluble but also possess versatile biological activities such as antitumor, immunostimulating, antioxidant and antimicrobial activity1). This study investigated antioxidative effects of carboxymethyl chitooligosaccharides (CM-COS) on the generation of direct free radical scavenging activity by using electron spin resonance (ESR) spectrometer and measure Fourier transform infrared (FT-IR) spectroscopy. Antioxidative activities of CM-COS at 70% and 80% were measured, respectively. And cytotoxic effect was measured by the method of Park et al1). In the results of ESR spectrometer, the CM-COS 80% had the highest antioxidant activity compared to the other CM-COS. The IC50 values of 80% CM-COS were 3.29, 6.25, 2.65 and 1.50 mg/ml for DPPH, hydroxly, superoxide, alkyl radical, respectively. Moreover, CM-COS did not show any cytotoxic effect against MRC-5 cell lines.

To improve the functional properties of chitosan, carboxymehtylchitosan (CM-chitosan) was synthesized by chemical modifications. CM-chitosan, an important water soluble chitosan derivative, has many attractive chemical, physical and biological properties such as gel-forming capability, low toxicity, and good biocompatibility, all of which make it a promising biomaterial1). In this study, we compared CM-chitosan with the other emulsifiers to emulsifying activity, and the optimum conditions for high emulsifying activity and stability of CM-chtiosan were investigated. Emulsifying activity of the CM-chitosan was investigated in various conditions such as concentration, water-oil ratios, pH and temperature. The emulsifying activity of CM-chitosan was 20% higher than of gelatin, bovine serum albumin, and Tween-80 as commercial emulsifier. The emulsifying activity of CM-chitosan solution was the highest at 1.5%, and was decreased with increasing concentration at above 1.5% of CM-chitosan. The optimum conditions of water-oil ratios. pH and temperature were 50% (w/v), pH 8.0 and 20℃, respectively. In these conditions, the emulsifying activity of CM-chitosan solution was 86%. The emulsion stability of CM-chitosan was higher than that of other commercial emulsifier. Moreover, CM-chitosan did not show any cytotoxic effect against MRC-5 cell lines.

Functionality of developed multi functional wellbeing tea was investigated, such as antioxidative capacity, anti-angiogenic effect, in vivo test, immunity increasing effect, H2O2 toxicity effect, anti-obesity test, blood analysis, testosteron induction effect and anti-coagulating effect. For anti-angiogenic effect, this wellbeing tea showed higher positive effect, it's very useful for anti-cancer and anti-obesity effect. From blood analysis, this tea strongly supressed GOT and GPT, and also expressed a novel decreasing effect of triglyceride. Especially KAP is showed for higher induction effect of testosteron. This multifunctional wellbeing tea could be used for elevating body functionality.

Natural products such as T.officinale. A.radix, Greentea, C.militaris, Kapi, G. pentaphyllum, WGS, C. coetex and T.veronicoides which born higher antioxidative capacity and anti-angiogenic effect, improving liver function and immunity, indicing testosteron effect were utilized to making multifunctional wellbeing tea. Some natural products which showed bitter and astringent taste were masked for more soften taste. For masking method, roasting, soaking and grinding were used for elevating their functions. And combination ratio of materials is also very important for multifunction and improving positive effects. The final combination for multifunctional wellbeing tea was Greentea: T.officinale : C.militaris : A.radix : G. pentaphyllum : WGS : KAP =1:3:1.5:2:1:1:1.5.

Some of natural products have been long used as a botanical medicine for anti-inflammation in korea. Inflammation is the complex biological response of vascularized living tissue to local injury. The function of inflammation is to enclose injury, destroy invading microorganisms and inactivate toxins. The process of inflammation, however, can potentially be harmful, such as hypersensitive reactions, chronic inflammation and progressive organ damage. Macrophages are considered to play essential roles in inflammation. When activated by endotoxin, macrophages produce Nitric oxide. The inhibitory effects of these natural products on NO production from lipopolysaccharide induced RAW 264.7 macrophages were measured. Among them, natural products with potent inhibition of NO production were selected, and then iNOS expression was measured using RT-PCR. The goals of this research were to study the anti-inflammatory effects of extracts from natural product in RAW 264.7 cell line.

This study was designed to examine the antioxidant activities of the essential oil and various extracts (hexane, chloroform, ethyl acetate and methanol) of Cestrum nocturnum L. Further, the total phenolic contents of the essential oil and extracts were determined, which was expressed as garlic acid equivalents (GAE). Ethyl acetate extract contained higher phenolic content (12.36 GAE mg/g) than other extracts. The antioxidative potential of the samples was evaluated by scavenging capacities of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide radicals. The extracts and essential oil displayed remarkable scavenging capacities of DPPH (IC50 range of 42.56-49.85 μg/ml) and superoxide radical (IC50 range of 18.37-19.26 μg/ml). Our findings demonstrate that the essential oil and various extracts of C. nocturnum possessed antioxidant activities that might be a natural potential source of preservative used in food and other allied industries.

There is intense research on natural antimicrobial derivatives from plant origin. The essential oil and the leaf extracts of M . glyptostroboides revealed remarkable antidermatophytic effect as a percentage of radial growth inhibition of 20.6 to 62.6% and 24.2 to 51.3%, respectively.

기존의 화학합성 방부제가 가지는 부작용을 해결하기 위해 최근 천연으로 제조된 독성의 염려가 없고 항균력이 있는 천연물의 자연친화적인 천연방부제의 연구가 활발히 이루어지고 있으며, 본 연구에서는 비자나무가 가지고 있는 약리효과(구충제사용 등)등의 기술개발 가능성을 활용하여 비자나무의 종자, 잎 추출물을 천연방부제로 개발 하였다. 실험재료로는 제주특별자치도 제주시 소재 산천단 인근에서 비자나무(종자, 잎)를 채취 및 파쇄, 이를 정유추출장치(모델명 : EssenLab-Plus, 용량 9L, 한일랩테크)및 별도 제작한 추출장치를 활용하여 각각의 정유 및 에탄올(70, 80, 90, 95%) 추출물을 확보하였다. 항균활성 실험을 수행하기 위하여 한국미생물보존센터에서 구입한 균주는 Staphylococcus aureus(12214), Staphylococcus epidermidis(40411), Streptococcus mutans(40105), Escherichia coli(41300) 및 Penicillium citrinum(11663), Aspergillus niger(32318)을 사용하여 Paper Disc법으로 자몽씨 추출물과 억제환의 크기를 비교평가하였다. 위 실험 결과 정유 추출물의 수득율은 잎에서 0.13%을 나타내었고, 에탄올 추출물 중에는 70, 80, 90, 95%추출물에서 종자(80%), 잎(70%) 각각의 수득율이 9.6%, 19.8%로 가장 높은 수득율을 보였다. 위의 Paper Disc 방법으로 6가지 균주에 대한 에탄올 추출물에서의 항균 활성을 확인할 수 있었으며, 특히 비자나무잎의 정유 추출물에서는 S. aureus 15.26mm, S. epidermidis 15.48mm의 활성 효과를 얻을 수 있었다.

The extraction conditions of fucoidan from Undaria pinnatifida sporophylls (UPS) were optimized using ultra-high pressure (UHP) process at 100 MPa. UHP treatment was carried out using different enzymes, particle sizes of UPS, extraction pH, and enzyme concentrations. Tunicase FN (β-glucanase) was selected based on recovery yield of fucoidan among various enzymes including glucanase, cellulase and pectinase. The recovery yield of fucoidan was the highest at pH 8 when Tunicase FN was used. Additionally the effects of particle size of raw material (UPS) on the recovery yield were investigated. In conclusion, optimal conditions for the extraction process of fucoidan from UPS using UHP enzyme treatment process were established.

Bacterial cellulose (BC) has recently gained much attention in the research realm for its potential applications in various fields, especially; their biomedical and cosmoceutical features are of considerable interest. These applications require BC in the film form, which can be produced only under static cultivation. The amount of BC produced in static conditions is lower compared to agitated/shaking culture conditions. In the present study, BC membranes were produced by G. hansenii PJK under static culture conditions in jar fermenters using chemically-defined medium in fed-batch cultivation. Fed-batch enhanced the BC production compared to the batch cultivation. The produced BC membranes were characterized using various techniques including field-emission scanning electron microscopy (FE-SEM), X-ray diffractometry (XRD), gel permeation chromatography (GPC), CP/MAS (13C-NMR), and Fourier Transform Infrared Spectroscopy (FTIR). The physico-mechanical characteristics of BC membranes were investigated by measuring its water holding capacity, water release rate and mechanical strength (stress-strain and elasticity). These results of production and characterization of BC membranes obtained in fed-batch cultivation will be presented in comparison to batch cultivation.

Bacterial Cellulose (BC) produced by numerous different techniques. The choice of a cultivation technique is strictly dependent on the biopolymer commercial destination. In the present study, BC was produced by G. hansenii PJK strain in shaking cultivation using the waste from beer fermentation broth (WBFB) as an alternative of chemically defined medium. The WBFB broth was treated with autolysis, hydrolysis and sonication. The supernatant of the resulting autolysed, hydrolysed and sonicated product were analyzed for elemental composition. The untreated WBCB was used as control system. The production yield of untreated WBFB was higher than the autolysed, hydrolysed and sonicated. The amount of BC produced was 2.25, 0.612, 0.124 and 1.65 g/L after 168 h using untreated, autolysed, hydrolysed, and sonicated WBCB, respectively. Adding 1% glucose (commercial grade) to the untreated, autolysed, hydrolyzed and sonicated WBCB improved the production of bacterial cellulose, 2.996, 0.516, 0.92 and 1.98 g/L, respectively. The by-product, water soluble oligosaccharide (WSOS) produced during the cultivation of BC increased from 4.5 to 6.5 g/L after 168 h of cultivation.