• Fisheries and Aquatic Sciences
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• v.14 no.4
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• pp.283-288
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• 2011

The jellyfish Nemopilema nomurai was hydrolyzed with papain and a novel dipeptide purified via ultrafiltration, gel filtration chromatography with Sephadex LH-20, and reverse phase chromatography using $C_{18}$ and $C_{12}$ columns. The IR, 1H NMR, 13C NMR, and MS spectrometer analyses showed that the dipeptide comprised tyrosine-isoleucine (Tyr-Ile). The $IC_{50}$ and $K_i$ values were $6.56{\pm}1.12$ and $3.10{\pm}0.28\;{\mu}M$, respectively, indicating competitive inhibition of angiotensin-I-converting enzyme (ACE). As a novel ACE-inhibitory active peptide, Tyr-Ile may have potential for use in antihypertensive therapy.

• Journal of Microbiology and Biotechnology
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• v.33 no.12
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• pp.1625-1634
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• 2023

Leuconostoc lactis strain DMLL10 was isolated from kimchi, a fermented vegetable, as a starter candidate through safety and technological assessments. Strain DMLL10 was susceptible to ampicillin, chloramphenicol, clindamycin, erythromycin, gentamicin, kanamycin, streptomycin, and tetracycline. It did not show any hemolytic activity. Regarding its phenotypic results related to its safety properties, genomic analysis revealed that strain DMLL10 did not encode for any toxin genes such as hemolysin found in the same genus. It did not acquire antibiotic resistance genes either. Strain DMLL10 showed protease activity on agar containing NaCl up to 3%. The genome of DMLL10 encoded for protease genes and possessed genes associated with hetero- and homo-lactic fermentative pathways for lactate production. Finally, strain DMLL10 showed antibacterial activity against seven common foodborne pathogens, although bacteriocin genes were not identified from its genome. These results indicates that strain DMLL10 is a novel starter candidate with safety, enzyme activity, and bacteriocin activity. The complete genomic sequence of DMLL10 will contribute to our understanding of the genetic basis of probiotic properties and allow for assessment of the effectiveness of this strain as a starter or probiotic for use in the food industry.

• Advances in nano research
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• v.2 no.2
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• pp.89-98
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• 2014

A novel and simple method for the synthesis of nano-magnetite particles is disclosed. In the novel procedure, $Fe^{2+}$ is the only source of metal cation. Carboxymethylcellulose (CMC) is used as the structure directing agent. The phase analysis of the nano-particles was performed using XRD and electron diffraction techniques. Size and morphology analysis was performed using light scattering and TEM techniques. The effect of $NH_4OH$ solution (32 wt. %) at different CMC concentrations on the size distribution of the final magnetite powders is studied. An optimal base concentration exists for each CMC concentration leading to minimal agglomeration. There exists a minimum CMC concentration (0.0016 wt. %), lower than that no magnetite forms. It is shown that using the new method, it is possible to immobilize a lipase enzyme (Candida Rugosa) with immobilization efficiency larger than 98 % with a loading more than 3 times the reported value in the literature. The latter phenomenon is explained based on the agglomerate state of the nano-particles in the liquid phase.

• YAKHAK HOEJI
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• v.36 no.2
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• pp.167-172
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• 1992

Klebsiella K-36 producing novel sulfotransferase was isolated from rat intestinal flora. The novel sulfotransferase catalyzed the transfer of sulfate group from p-nitrophenylsulfate to phenolic compounds but it did not use PAPS(3'-phosphoadenosine 5'-phosphosulfate) as a donor substrate. The present enzyme was 160 K daltons. Optimal pH was 10. When p-nitrophenyl sulfate was used as a donor substrate, 1-naphthol was the best substrate, followed by phenol, phenanthrol and tyrosine. The apparent Km for phenol using p-nitrophenylsulfate as a donor substrate and that for p-nitrophenylsulfate using phenol as an acceptor substrate were determined to be 0.66 mM and 0.11 mM, respectively.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1549-1558
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• 2017

Despite significant efforts to improve the treatment of tuberculosis (TB), it remains a prevalent infectious disease worldwide owing to the limitations of current TB therapeutic regimens. Recent work on novel TB treatment strategies has suggested that directly targeting host factors may be beneficial for TB treatment. Such strategies, termed host-directed therapeutics (HDTs), focus on host-pathogen interactions. HDTs may be more effective than the currently approved TB drugs, which are limited by the long durations of treatment needed and the emergence of drug-resistant strains. Targets of HDTs include host factors such as cytokines, immune checkpoints, immune cell functions, and essential enzyme activities. This review article discusses examples of potentially promising HDTs and introduces novel approaches for their development.

• BMB Reports
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• v.40 no.6
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• pp.911-920
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• 2007

Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the leading infectious diseases to humans. It is urgent to discover novel drug targets for the development of antitubercular agents. The 2-C-methyl-Derythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has been considered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammals. MEP cytidyltransferase (IspD), the third-step enzyme of the pathway, catalyzes MEP and CTP to form 4-diphosphocytidyl-2-C-methylerythritol (CDP-ME) and PPi. In the work, ispD gene from M. tuberculosis H37Rv (MtIspD) was cloned and expressed. With N-terminal fusion of a histidine-tagged sequence, MtIspD could be purified to homogeneity by one-step nickel affinity chromatography. MtIspD exists as a homodimer with an apparent molecular mass of 52 kDa. Enzyme property analysis revealed that MtIspD has high specificity for pyrimidine bases and narrow divalent cation requirements, with maximal activity found in the presence of CTP and $Mg^{2+}$. The turnover number of MtIspD is $3.4 s^{-1}$. The Km for MEP and CTP are 43 and $92{\mu}M$, respectively. Furthermore, MtIspD shows thermal instable above $50^{\circ}C$. Circular dichroism spectra revealed that the alteration of tertiary conformation is closely related with sharp loss of enzyme activity at higher temperature. This study is expected to help better understand the features of IspD and provide useful information for the development of novel antibiotics to treat M. tuberculosis.

• Journal of Microbiology and Biotechnology
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• v.18 no.9
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• pp.1555-1563
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• 2008

A novel $\alpha$-amylase ($\alpha$-1,4-$\alpha$-D-glucan glucanohydrolase, E.C. 3.2.1.1), ScAmy43, was found in the culture medium of the phytopathogenic fungus Sclerotinia sclerotiorum grown on oats flour. Purified to homogeneity, ScAmy43 appeared as a 43 kDa monomeric enzyme, as estimated by SDS-PAGE and Superdex 75 gel filtration. The MALDI peptide mass fingerprint of ScAmy43 tryptic digest as well as internal sequence analyses indicate that the enzyme has an original primary structure when compared with other fungal a-amylases. However, the sequence of the 12 N-terminal residues is homologous with those of Aspergillus awamori and Aspergillus kawachii amylases, suggesting that the new enzyme belongs to the same GH13 glycosyl hydrolase family. Assayed with soluble starch as substrate, this enzyme displayed optimal activity at pH 4 and $55^{\circ}C$ with an apparent $K_m$ value of 1.66 mg/ml and $V_{max}$ of 0.1${\mu}mol$glucose $min^{-1}$ $ml^{-1}$. ScAmy43 activity was strongly inhibited by $Cu^{2+}$, $Mn^{2+}$, and $Ba^{2+}$, moderately by $Fe^{2+}$, and was only weakly affected by $Ca^{2+}$ addition. However, since EDTA and EGTA did not inhibit ScAmy43 activity, this enzyme is probably not a metalloprotein. DTT and $\beta$-mercaptoethanol strongly increased the enzyme activity. Starting with soluble starch as substrate, the end products were mainly maltotriose, suggesting for this enzyme an endo action.

• BMB Reports
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• v.38 no.6
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• pp.633-638
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• 2005

Biosynthesis of prostanoids is regulated by three sequential enzymatic steps, namely phospholipase $A_2$ enzymes, cyclooxygenase (COX) enzymes, and various lineage-specific terminal prostanoid synthases. Prostaglandin E synthase (PGES), which isomerizes COX-derived $PGH_2$ specifically to $PGE_2$, occurs in multiple forms with distinct enzymatic properties, expressions, localizations and functions. Two of them are membrane-bound enzymes and have been designated as mPGES-1 and mPGES-2. mPGES-1 is a perinuclear protein that is markedly induced by proinflammatory stimuli, is down-regulated by anti inflammatory glucocorticoids, and is functionally coupled with COX-2 in marked preference to COX-1. Recent gene targeting studies of mPGES-1 have revealed that this enzyme represents a novel target for anti-inflammatory and anti-cancer drugs. mPGES-2 is synthesized as a Golgi membrane-associated protein, and the proteolytic removal of the N-terminal hydrophobic domain leads to the formation of a mature cytosolic enzyme. This enzyme is rather constitutively expressed in various cells and tissues and is functionally coupled with both COX-1 and COX-2. Cytosolic PGES (cPGES) is constitutively expressed in a wide variety of cells and is functionally linked to COX-1 to promote immediate $PGE_2$ production. This review highlights the latest understanding of the expression, regulation and functions of these three PGES enzymes.

• Journal of Microbiology and Biotechnology
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• v.24 no.5
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• pp.719-723
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• 2014

Caspases are a family of cysteine proteases that play an important role in the apoptotic pathway. Caspase-6 is an apoptosis effector that cleaves a variety of cellular substrates. The active form of the enzyme is required for use in research. However, it has been difficult to obtain sufficient quantities of active caspase-6 from Escherichia coli. In the present study, we constructed a caspase-6 with a 23-amino-acid deletion in the pro-domain. This engineered enzyme was expressed as a soluble protein in E. coli and was purified using affinity resin. In vitro enzyme assay and cleavage analysis revealed that the engineered active caspase-6 protein had characteristics similar to those of wild-type caspase-6. This novel method can be a valuable tool for obtaining active caspase-6 that can be used for screening caspase-6-specific substrates, which in turn can be used to elucidate the function of caspase-6 in apoptosis.

• Fibers and Polymers
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• v.6 no.1
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• pp.1-5
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• 2005

In this study, we attempted to evaluate a novel use of sericin-fixed silk fiber (SFx) as an immobilization support of enzyme. Sericin was fixed on the silk fiber using glutaraldehyde as a fixation reagent. After 6 hours of fixation, the degree of fixation increases linearly with linear decrease of the amount of bound $\alpha$-chymotrypsin (CT). This suggests that the increase of the degree of fixation is due to the further crosslinking of free aldehyde groups on the surface of sericin-fixed silk fiber (SFx). Even though perfect fixation was not achieved, sericin did not dissolve seriously and could be removed by further washing. The specific activity did not differ significantly after 6 hours of fixation. The activity of immobilized CT on SFx decreased to its half after 6 hours of incubation at 50$^{\circ}C$. However, it retained $78\%$ of initial activity even after 1 hour of treat­ment with $100\%$ ethanol. As a result, the SFx could be used as an immobilization support of enzyme in non-aqueous media at ambient temperature.