• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1660-1669
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• 2015

The present study describes the gene cloning, overexpression and characterization of a novel nitrilase from hyperthermophilic bacterium Thermotoga maritima MSB8. The nitrilase gene consisted of 804 base pairs, encoding a protein of 268 amino acid residues with a molecular mass of 30.07 kDa after SDS-PAGE analysis. The optimal temperature and pH of the purified enzyme were 45℃ and 7.5, respectively. The enzyme demonstrated good temperature tolerance, with 40% residual activity after 60 min of heat treatment at 75℃. The kinetic constants V_max and K_m of this nitrilase toward 3-cyanopyridine were 3.12 μmol/min/mg and 7.63 mM, respectively. Furthermore, this novel nitrilase exhibited a broad spectrum toward the hydrolysis of the aliphatic nitriles among the tested substrates, and particularly was specific to aliphatic dinitriles like succinonitrile, which was distinguished from most nitrilases ever reported. The catalytic efficiency k_cat/K_m was 0.44 /mM/s toward succinonitrile. This distinct characteristic might enable this nitrilase to be a potential candidate for industrial applications for biosynthesis of carboxylic acid.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.109-114
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• 1997

The Oomycete Saprolegnia ferax produces an extracellular $\beta$-amylase, Maximum enzyme yield was attained after 7 days of growth in YNB starch medium (pH 6.5) at 25$\circ$C. The amylase was pu- rified 24-fold by ultrafitration, HPLC DEAE column and HPLC gel filtration. The purfied enzyme was a monomeric glycoprotein with a molecular weight of about 44,000 dalton. The pH and temperature optima were 6.5 and 50$\circ$C, respectively. The enzyme was fairly stable up to 50$\circ$C and at acidic pH region (pH 4.0-7.0). The apparent Km and Vmax values of the enzyme against soluble starch were 0.77 mg/ml and 2,174 $\mu$moles/mg protein, respectively. Amino acid analysis indicated that the enzyme was enriched in alanine, glycine, leucine and acidic amino acid. Starch hydrolysis with the enzyme released maltose but not glucose, whereas maltotriose, Schardinger dextrin ($\alpha$-cyclodextrin) and pullulan were not hydrolysed by the enzyme. The enzyme was inhibited by Schardinger dextrin, p-chloromercuribenzoate(PCMB), CU$^{2+}$' and Hg$^{2+}$. Inhibition of the enzyme by PCMB could be reversed by the addition of cysteine and mercaptoethanol.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.417-430
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• 2003

A new method was developed to prepare microcapsules involving hydrophobic components. A totally new "silicone-resin-polypeptide" was used as the wall materials. The polypeptide was made by hydrolysis of collagen and silk protein and so on, and that was combined with silicone. This microcapsule was easily prepared from silicone-resin-polypeptide in water solution. The ratio of encapsulation in the microcapsule was not only high level as 90％, which had never been reached, but also the particle size could be controlled to obtain very small size (average particle size: 2${\mu}{\textrm}{m}$). Moreover, these microcapsules were resistant to high shearing forces and were stable over a long time period. This stable microcapsule was not crushed in pressure with finger spreading, so the core materials hardly touch the skin directly. Application in cosmetics by using microcapsule involving UV absorbents (2-ethylhexy1-4-methoxycinnamate (OMC) and 4-tert-butyl-4' -methoxydibenzoyl-methane (BMDBM)) was examined. It was possible to apply organic UV absorbents in water-rich formulations without any surfactant by using this microcapsule. This formulation demonstrated a good moisturizing and soft skin feel. Therefore, the microcapsule was applied to hair care products. As a result, the sunscreen hair lotion with microcapsule was able to prevent from damaging and decoloring of hair color by UV rays. As just, it was suggested that this microcapsules were be widely applied in cosmetics.cosmetics.

• Fisheries and Aquatic Sciences
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• 제5권4호
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• pp.311-313
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• 2002

Proteolytic activity of live Uronema manum was analyzed by fluorescence polarization (FP) technique. Protease activity was measured by a decrease in FP value using fluorescein isothiocynate (FITC)-casein as a protein substrate. The results demonstrated an inverse linear relationship between fluorescence polarization (FP) values and live ciliate concentration over the range $1\times10^4\;to\;2\times10^5$ cells/well. However, the FP values of $10-10^3$ live parasites were not different significantly from that of control. Time-dependent decrease in FP value was shown in the wells containing live U. marinum. In the present study, FP assay had the benefit to provide measurements of substrate hydrolysis by live parasites in real-time, and did not require separations, precipitations, or transfers of reaction mixture.

• 한국환경과학회지
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• 제16권4호
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• pp.409-414
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• 2007

Effects of dimethipin on ${\alpha}$-amylase activity of barley seeds were investigated. In the treatments of $1{\mu}M\;and\;10{\mu}M$ dimethipin, the indexes of germination were reduced to 17% and 24 % respectively. After seed germination, dimethipin was added to germinated seedlings and then the seedlings were kept to measure seedling length under illumination for 7 days. In control, the length of seedling was 5.7 cm, but in the treatments of $1{\mu}M$ dimethipin and $10{\mu}M$ dimethipin, seedling lengths were 5.5 cm and 1.2 cm respectively. In the relationship between dimethipin concentrations and ${\alpha}$-amylase activities, there was a linear curve. The more dimethipin was added to the seeds, the more ${\alpha}$-amylase activities were inhibited. In the treatments of $1{\mu}M$ dimethipin and $10{\mu}M$ dimethipin, ${\alpha}$-amylase activities were reduced to 33% and 71% respectively. Dimethipin also inhibited ${\alpha}$-amylase activities increased by gibberellin and the content of soluble protein. Therefore, it could be suggested that dimethipin might inhibit directly the activities of hydrolysis enzymes including ${\alpha}$-amylase or the expression of ${\alpha}$-amylase genes as germination and seedling growth were severely disturbed.

• Preventive Nutrition and Food Science
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• 제16권4호
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• pp.339-347
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• 2011

Flour and isolated starch from chickpea (desi type, 328S-8) were evaluated for their in vitro digestibility and physicochemical properties. The protein content, total starch content and apparent amylose content of chickpea flour and isolated starch were 22.2% and 0.6%, 45.8% and 91.5%, and 11.7% and 35.4%, respectively. Chickpea starch granules had an oval to round shape with a smooth surface. The X-ray diffraction pattern of chickpea starch was of the C-type and relative crystallinity was 24.6%. Chickpea starch had only a single endothermic transition (13.3 J/g) in the DSC thermogram, whereas chickpea flour showed two separate endothermic transitions corresponding to starch gelatinization (5.1 J/g) and disruption of the amylose-lipid complex (0.7 J/g). The chickpea flour had a significantly lower pasting viscosity without breakdown due to low starch content and interference of other components. The chickpea starch exhibited significant high setback in the viscogram. The average branch chain length, proportion of short branch chain (DP 6~12), and long branch chains (DP${\geq}$37) of isolated chickpea starch were 20.1, 20.9% and 9.2%, respectively. The rapidly digestible starch (RDS), slowly digestible starch (SDS) and resistant starch (RS) contents of chickpea flour and starch were 9.9% and 21.5%, 28.7% and 57.7%, and 7.1% and 9.3%, respectively. The expected glycemic index (eGI) of chickpea flour (39.5), based on the hydrolysis index, was substantially lower than that of isolated chickpea starch (69.2).

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권1호
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• pp.18-24
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• 2001

The advantage of using Streptomyces griseus HUT 6037 in the production of chitinase or chitosanase is that the organism is capable of hydrolyzing amorphous or crystalline chitin and chitosan according to the type of the substrate used. We investigated the effects of the enzyme induction time and chitin sources, CM-chitosan and deacetylated chitosan (degree of deacetylation 75-99%), on production of chitosanase. We found that this strain accumulated chitosanase when cells were grown in the culture medium containing chitosanaceous substrates instead of chitinaceous substrates. The highest chitosanase activity was obtained at 4 dyas of cultivation with 99% deacetylated chitosan. The specific activities of chitinase and chitosanase were 0.91 and 1.33 U/mg protein at 3 and 5 days, respectively. From the study of the enzymatic digestibility of various degrees of deacetylated chitosan, it was found that (GlcN)$_3$, (GlcN)$_4$and (GlcN)(sub)5 were produced during the enzymatic hydrolysis reaction. The results of this study suggested that the sugar composition of (GlcN)$_3$was homogeneous and those of (GlcN)$_4$and (GlcN)(sub)5 were heterogeneous.

• BMB Reports
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• 제34권5호
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• pp.457-462
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• 2001

Using the monomolecular film technique, we compared the interfacial properties of Staphylococcus simulans lipase (SSL) and Staphylococcus aureus lipase (SAL). These two enzymes act specifically on glycerides without any detectable phospholipase activity when using various phospholipids. Our results show that the maximum rate of racemic dicaprin (rac-dicaprin) hydrolysis was displayed at pH 8.5, or 6.5 with Staphylococcus simulans lipase or Staphylococcus aureus lipase, respectively The two enzymes interact strongly with egg-phosphatidyl choline (egg-PC) monomolecular films, evidenced by a critical surface pressure value of around $23\;mN{\cdot}m^{-1}$. In contrast to pancreatic lipases, $\beta$-lactoglobulin, a tensioactive protein, failed to inhibit Staphylococcus simulans lipase and Staphylococcus aureus lipase. A kinetic study on the surface pressure dependency, stereoselectivity, and regioselectivity of Staphylococcus simulans lipase and Staphylococcus aureus lipase was performed using optically pure stereoisomers of diglycerides (1,2-sn-dicaprin and 2,3-sn-dicaprin) and a prochiral isomer (1,3-sn-dicaprin) that were spread as monomolecular films at the air-water interface. Both staphylococcal lipases acted preferentially on distal carboxylic ester groups of the diglyceride isomer (1,3-sn-dicaprin). Furthermore, Staphylococcus simulans lipase was found to be markedly stereoselective for the sn-3 position of the 2,3-sn-dicaprin isomer.

• 한국식품조리과학회지
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• 제7권4호
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• pp.93-101
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• 1991

The object of this study was to investigate characteristics of kiwifruit protesae and effect of this enzyme on the functionality of casein. The specific activity of crudely prepared kiwifruit pretense on casein was 196.95 units/mg protein, it showed optimum activity at pH 3.0, $60^{\circ}C$. The degree of hydrolysis of casein with pretense treatment steeply increased to 73.5% and 78.9% for 10 and 20 minutes and then reached 84.1% and 89.3% for 1 and 4 hours, respectively. Solubility of non heated control group was 0.2% at pH 4, while the sample groups treated with enzyme for 0, 10 and 20 minutes were 14.5%, 19.2% and 24.0%, respectively. Casein treated with pretense showed marked increase in foam expansion near isoelectric point. However, enzymatically treated groups had lower foam expansion than the control groups. Foam stabilities of enzymatically modified group were lower than those of the control groups at all pH. Emulsifying activity of the non-heated control group was 0% at pH 4, while the groups modified enzymatically for 0, 10, and 60 minutes showed 51.0%, 55.5% and 54.5%, respectively.

• 한국축산식품학회지
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• 제27권1호
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• pp.102-109
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• 2007

Lactobacillus salivarius subsp. salivarius CNU27 possessed a high level of ${\alpha}$-galactosidase activity. Purified ${\alpha}$-galactosidase was obtained after sonication of harvested cell pellet followed by DEAE-Sephadex A-50 and Mono Q anion exchange chromatography. The specific activity of the purified enzyme was 8,994 units/mg protein which is 17.09 times higher than that in crude extract. The native enzyme was a monomer with a molecular mass of 56,397.1 dalton. The optimum temperature and pH for the enzyme were $40^{\circ}C$ and 6.0, respectively. The enzyme was stable between 25 and $50^{\circ}C$. However, ${\alpha}$-galactosidase activity was lost rapidly below pH 4.5 and above pH 8.5. The enzyme activity decreased to 6.73% and 4.30% of the original activity by addition of $Cu^{2+}$ and $Hg^{2+}$, respectively. Other metal compounds did not affect the enzyme activity significantly. The enzyme liberated galactose from melibiose, raffinose, and stachyose. The rate of substrates hydrolysis was measured by HPLC. Raffinose, stachyose and melibiose were completely decomposed after 24 hr at $40^{\circ}C$.