• Food Science and Biotechnology
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• v.16 no.5
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• pp.843-847
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• 2007

Strain NK34 was characterized for probiotic use. Strain NK34 was named Lactococcus lactis NK34 based on API 50 CHL kit results and 16S rDNA sequencing. L. lactis NK34 was highly resistant to artificial gastric juice (pH 2.5) and artificial bile acid. Based on results from the API ZYM kit, 4 enzymes were produced. L. lactis NK34 was resistant to all antibiotics tested except for $10\;{\mu}g/mL$ roxithromycin and $10\;{\mu}g/mL$ erythromycin. The cholesterol-lowering effect of L. lactis NK34 was about 46.9%. Concentrations of interleukin $(IL)-1{\alpha}$ in the $20{\times}$ concentrated supernatant of L. lactis NK34 was about 361 pg/mL. L. lactis NK34 was also found to inhibit the growth of colon cancer cells due to MNNG-induced DNA damage. These results demonstrate the potential of L. lactis NK34 as a health-promoting probiotic.

• BMB Reports
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• v.40 no.4
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• pp.494-500
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• 2007

The endophytic bruchid pest Callosobruchus maculatus causes severe damage to storage cowpea seeds, leading to economical losses. For this reason the use of $\alpha$-amylase inhibitors to interfere with the pest digestion process has been an interesting alternative to control bruchids. With this aim, $\alpha$-amylase inhibitors from baru seeds (Dipteryx alata) were isolated by affinity chromatographic procedures, causing enhanced inhibition of C. maculatus and Anthonomus grandis $\alpha$-amylases. To attempt further purification, this fraction was applied onto a reversed-phase HPLC column, generating four peaks with remarkable inhibition toward C. maculatus $\alpha$-amylases. SDS-PAGE and MALDI-ToF analysis identified major proteins of approximately 5.0, 11.0, 20.0 and 55 kDa that showed $\alpha$-amylase inhibition. Results of in vivo bioassays using artificial seeds containing 1.0% (w/w) of baru crude extract revealed 40% cowpea weevil larvae mortality. These results provide evidence that several $\alpha$-amylase inhibitors classes, with biotechnological potential, can be isolated from a single plant species.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.6
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• pp.1519-1523
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• 2003

Glehnia littoralis Fr. Schmidt (Umbelliferae) is a short-lived herbaceous species that are mostly distributed throughout East Asia. Although G. littoralis has been regarded as ecologically important one, there is no report on population structure in Korea. Starch gel electrophoresis was used to investigate the allozyme variation and genetic structure of Korean populations of this species. A high level of genetic variation was found in G. littoral is populations. Nine enzymes revealed 18 loci, of which 12 were polymorphic (66.7%). Genetic diversity at the species and population levels were 0.159 and 0.129, respectively. The sexual and asexual reproduction, high fecundity, and colonization process are proposed as possible factors contributing to genetic diversity. An indirect estimate of the number of migrants per generation (Nm = 1.45) indicated that gene flow was not extensive among Korean populations of this species. It is suggested that the ability of vegetation and artificial selection may have played roles in shaping the population structure of this species. we recommend that a desirable conservation population should be included at least 30 plants per population and especially those with high variation.

• BMB Reports
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• v.39 no.4
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• pp.464-467
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• 2006

Recently, the nucleotide sequences of entire genomes became available. This information combined with older sequencing data discloses the exact chromosomal location of millions of nucleotide markers stored in the databases at NCBI, EMBO or DDBJ. Despite having resolved the intron/exon structures of all described genes within these genomes with a stroke of a pen, the sequencing data opens up other interesting possibilities. For example, the genomic mapping of the end sequences of the human, murine and rat BAC libraries generated at The Institute for Genomic Research (TIGR), reveals now the entire encompassed sequence of the inserts for more than a million of these clones. Since these clones are individually stored, they are now an invaluable source for experiments which depend on genomic DNA. Isolation of smaller fragments from such clones with standard methods is a time consuming process. We describe here a reliable one-step cloning technique to obtain a DNA fragment with a defined size and sequence from larger genomic clones in less than 48 hours using a standard vector with a multiple cloning site, and common restriction enzymes and equipment. The only prerequisites are the sequences of ends of the insert and of the underlying genome.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.59-64
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• 2002

An angiotensin I-converting enzyme (EC 3.4.15.1) (ACE), which can convert inactive angiotensin I into angiotensin II, a vasoconstrictor, is one of the key enzymes in controlling hypertension. It is suggested that the inhibition of ACE prevents hypertension, and many inhibitory peptides have already been reported. In the current study, oligonucleotides encoding ACE inhibitory peptides (IY, VKY) were chemically synthesized and designed to be multimerised due to isoschizomer sites (BamHI, BglII). The cloned gene named AP3 was multimerised up to 6 times in pBluescript and expressed in BL2l containing pGEX-KG. The fusion protein (GST-AP3) was easily purified with a high recovery by an affinity resin, yielding 38 mg of synthetic AP3 from a 1-1 culture. The digestion of AP3 by chymotrypsin exhibited an $IC_50$ value of $18.53{\mu}M$. In conclusion, the present experiment indicated that AP3 could be used as a dietary antihypertensive drug, since the potent ACE inhibitory activity of AP3 could be activated by chymotrypsin in human intestine.

• Food Science and Biotechnology
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• v.15 no.2
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• pp.227-231
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• 2006

Strain NK181 was isolated for probiotic use from jeotkal and based on results of API 50 CHL kit and 16S rDNA sequencing was tentatively named Lactobacillus plantarum NK181. L. plantarum NK181 was highly resistant to artificial gastric juice (pH 2.5) and bile acid and demonstrated strong adherence to Caco-2 cells. In test using API ZYM kit, eight enzymes were produced. Supernatant of L. plantarum NK181 exhibited about 30% 1,1-diphenyl-2-picyryl hedrazyl (DPPH) radical-scavenging activity and reduced cholesterol by 70%. These results demonstrate potential use of L. plantarum NK181 as health-promoting probiotic.

• Journal of Plant Biotechnology
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• v.4 no.2
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• pp.59-65
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• 2002

ADP-glucose pyrophosphorylase (AGPase) catalyzes the key regulatory step in starch biosynthesis. Two cDNA clones encoding AGPase subunits were isolated from the leaf cDNA library of Chinese cabbage (Brassica campestris L. spp. pekinensis). One was designated as BCAGPS for the small subunit and the other as BCAGPL for the large subunit. Both cDNAs have uninterrupted open reading frames deriving 57 kDa and 63 kDa polypeptides for BCAGPS and BCAGPL, respectively, which showed significant similarity to those of other dicot plants. Also, However, the deduced amino acid sequence of BCAGPL has a unique feature. That is, it contains two regions (Rl and R2) lacking in all other plant enzymes. This is the first report of BCAGPL containing Rl and R2 among plant large subunits as well as small subunits. From the genomic Southern analysis and BAC library screening, we inferred the genomic status of BCAGPS and BCAGPL gene.

• Food Science and Preservation
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• v.7 no.1
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• pp.1-7
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• 2000

To develop natural antimicrobial agents for keeping qualities of postharvested greenhouse produce the antimiocrobial actions of Polygonum cuspidatum Sieb. et Zucc. extract , which showed remarkable antimicrobial effects against microorganism causing the postharvest decay of greenhouse produce, were investigate. In the inhibitory experiment of enzymes related to energy production metabolism hexokinase activities decreased to 73% and 68% by treating with Polygonum cuspidatum Sieb. et.Zucc. extract and Eugenia caryophyllata Thumnberg extract in comparison with control, respectively. Direct visualization of microbial cells by using both transmission electron microscope and scanning electron microscope showed that microbial cell membrane was destroyed by treating with the dilute extract solution. this change of celluloar membrane permeability could be identified in the experiment that 0-nitrophenyl-${\beta}$-D-galactopyrano-side(ONPG), the artificial substrate of ${\beta}$-galactosidase, was hydrolyzed in the presence of the extract, indicating that the membrane was perturbed. The separation and identification of the most antimicrobialo substances isolated from Polygonum cuspidatum Sieb et. Zucc. extract and Eugenia caryophyllata Thunberg extract were carried out by using gas chromatography and mass spectrometry 9GC/MSD), which were identified as eugenol. As a result, the functionality of Polygonum cuspidatum Sieb. et Zucc. extract and Eugenia caryophyllata Thunberg extract as antimicrobial agents for keeping qualities of postharvested greenhouse produce may be recommended.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.199-207
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• 2002

The tropane alkaloids hyoscyamine (its racemic form being atropine) and scopolamine are used medicinally as anticholinergic agents that act on the parasympathetic nerve system. Because they differ in their actions on the central nervous system, currently there is a 10-fold higher commercial demand for scopolamine, in the N-butylbromide form, than there is for hyoscyamine and atropine combined. Several solanaceous species have been used as the commercial sources of these alkaloids, but the scopolamine contents in these plants often are much lower than those of hyoscyamine. For this reason there has been long-standing interest in increasing the scopolamine contents of cultivated medicinal plants. Naturally occurring and artificial interspecific hybrids of Duboisia have high scopolamine contents and are cultivated as a commercial source of scopolamine in Australia and other countries. Anther culture combined with conventional interspecific hybridization also has been used to breed high scopolamine-containing plants in the genera Datura and Hyoscyamus, but without much success. The use of recombinant DNA technology for the manipulation of metabolic processes in cells promises to provide important contributions to basic science, agriculture, and medicine. In this review, I introduce on the enzymes and genes involved in tropane alkaloid biosynthesis and current progress in metabolic engineering approaches for tropane alkaloid, especially scopolamine, production.

• Journal of Microbiology and Biotechnology
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• v.32 no.2
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• pp.205-211
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• 2022

Probiotics can effectively modulate host immune responses and prevent gastrointestinal diseases. The objective of this study was to investigate the probiotic characteristics of Lactobacillus brevis KU15152 isolated from kimchi and its protective potential against intestinal inflammation induced by Staphylococcus aureus lipoteichoic acid (aLTA). L. brevis KU15152 exhibited a high survival rate in artificial gastric and bile environments. Additionally, the adhesion capability of the strain to HT-29 cells was higher than that of L. rhamnosus GG. L. brevis KU15152 did not produce harmful enzymes, such as β-glucuronidase, indicating that it could be used as a potential probiotic. The anti-inflammatory potential of L. brevis KU15152 was determined in HT-29 cells. Treatment with L. brevis KU15152 suppressed the production of interleukin-8 without inducing significant cytotoxicity. The downregulatory effects of L. brevis KU15152 were involved in the suppression of nuclear factor-kappa B activation mediated by the extracellular signal-regulated kinase and Akt signaling pathways. Collectively, these data suggest that L. brevis KU15152 can be used in developing therapeutic and prophylactic products to manage and treat aLTA-induced intestinal damage.