• Toxicological Research
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• v.11 no.1
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• pp.23-29
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• 1995

Microsomes from human liver sample HL 110 oxidized aflatoxin $B_1$ $(AFB_1)$ to $AFB_1$ exo-8, 9-epoxide which was detected as a glutathione (GSH) conjugate with excess GSH S-transferase and to aflatoxin $Q_1$ ($AFB_1$; 3$\alpha$-hydroxyafiatoxin $B_1$), and testosterone to 6$\beta$-hydroxytestosterone. Anti-P450 3A4 nearly completely inhibited all of the reactions. Some fiavonoids inhibited all of the reactions. While other fiayonolds stimulated 8, 9-epoxidation and inhibited 3$\alpha$-hydroxylation. Gestodene inhibited all of the reactions when gestodene was metabolized by human liver microsomal P450 3A4 prior to adding substrate. But, ges-todene was added in the enzyme mixtures in the presence of $AFB_1$, it could not inhibit 8, 9-epoxidation of $AFB_1$. Nifedipine and troleandomycin inhibited both of the reactions of $AFB_1$ but only 3$\alpha$-hydroxylation was inhibited by the oxidation product of nifedipine. Although, troleandomycin was known as a mechanism-based inhibitor, the chemical did not show any detectable inhibitory effect on 6$\beta$-hydroxylation of testosterone. The results suggest that there are several different substrate-binding sites on P450 3A4.

• The Korean Journal of Mycology
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• v.22 no.4
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• pp.286-297
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• 1994

The optimum nutritional and cultural conditions of polygalacturonase by Ganoderma lucidum in liquid culture were studied. The optimal temperature, pH, and the duration of culture for production of the enzyme was $30^{\circ}C$, 5.5 and 14 days, respectively. The maximal production of the enzyme was obtained in a synthetic medium containing 10 g of pectin, 10 g of soluble starch, 1 g of yeast extract, 2 g of peptone, 1 g of phenylalanine, 2 g of $KH_2PO_4$, 0.2 g of $MgSO_4{\cdot}7H_2O$, 0.05 g of $CaCI_2$ and 100 g of $thiamin{\cdot}HCI$ in 1000 ml of distilled water.

• Fisheries and Aquatic Sciences
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• v.26 no.6
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• pp.393-405
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• 2023

The aim of this research was to enhance the flavor of visceral extracts from skipjack tuna. Flavor precursors and the optimum condition for the Maillard reaction were determined. The flavor extract was prepared from the tuna viscera using Endo/Exo Protease controlled in 3 factors; temperature, enzyme amounts and incubation time. The optimal condition for producing tuna viscera protein hydrolysate (TVPH) was 60℃, 0.5% enzyme (w/w) and 4-hour incubation time. TVPH were further processed to tuna viscera flavor enhancer (TVFE) with Maillard reaction. The Maillard reactions of TVFE were conducted with or without supplements such as xylose, yeast extract and methionine. The Maillard volatile components were analyzed with gas chromatography-mass spectrometry. Sixteen volatiles such as 2-methylpropanal, methylpyrazine, 2,5-dimethylpyrazine, dimethyl disulfide and 2-acetylthaizone were newly formed via Maillard reaction and the similarity of volatile contents from TVPH and TVFE were virtualized using Pearson's correlation integrated with heat-map and principal component analysis. To virtualize aromagram of TVPH and TVFE, odor activity value and odor impact spectrum (OIS) techniques were applied. According to OIS results, 3-methylbutanal, 2-methylbutanal, 1-octen-3-ol 2,5-dimethylpyrazine, methional and dimethyl trisulfide were the potent odorants contributed to the meaty, creamy, and toasted aroma in TVFE.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.415-422
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• 1996

A gene coding for a pyruvyl transferase enzyme involved in exopolysaccharide biosynthesis of Zoogloea ramigera 115SLR was isolated and sequenced. A 4.5 kb of BamHI DNA fragment was isolated from chromosomal DNA using a probe derived from ketal pyruvyl transferase gene of Xanthomonas campestris. The nucleotide sequence of 2.66 kb Pst1/HindIII DNA fragment which was homology with a probe revealed the existence of two complete open reading frames (ORF2 and ORF3) and two partial open reading frames (ORFI and ORF4). The deduced amino acid sequence of ORF3 was homologous to the ketalase (GumL product) of X campestris with 49.5% of similarity and 21.6% of identity. ORF2 on the other hand showed the higher identity with the ketalase (ExoV product) of Rhizobium meliloti (36%) as well as the ketalase of X campestris (23%) than that of ORF3. A gene product of ORF2 was determined with a bacteriophage T7 RNA polymerase/promoter system in E. coli. The molecular weight of protein was 33,500 dalton.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.229-233
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• 2001

Trichoderma reesei Rut C-30 produced high levels of ${\beta}$-glucosidase, endo-${\beta}$-glucosidase, endo-${\beta}$-1,4-glucanase, and exo-${\beta}$-1,4-glucanase. In pilot-scale production (50-1 fermentor), productivity and yield of CMCase (carborymethyl cellulose) and FPase (filter paper activity) were 273 U/ml and 35 U/ml, and 162 FPU/l.h and 437 FPU/g, respectively. The fed-batch techniques were used to improve enzyme activities with constant cell concentration. The acidity was an important parameter and controlled at pH 3.9 and 5.0 by automatic addition of ammonium hydroxide. Cellulase powder was prepared by ammonium sulfate precipitation and its CMCase and FPase activities were 3,631 U/g and 407 U/g, respectively.

• Korean Journal Plant Pathology
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• v.10 no.4
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• pp.277-283
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• 1994

An antimicrobial chitinase was purified from grapefruit extract and its properties were characterized. The chitinase was purified with a single step chromatography on regenerated chitin affinity gel column. The molecular weight of the purified chitinase was 29 kDa. The grapefruit extract contained the chitinase protein more than 50% of its total soluble proteins measured by coomassie stained protein bands. When the purified chitinase was incubated with polymers of N-acetylglucosamine (NAG), such as mycelia of Fusarium oxysproum and swollen chitin, they were degraded to oligosaccharides, and the oligosaccharides were then further hydrolyzed by the same enzyme to monomer and dimer of NAG. This result suggests that the chitinase contained both endo- and exo- chitinase activities. The chitinase was stable to heat and pH treatment; its activity was not diminished by the heat treatment upto 7$0^{\circ}C$ for 1 hr, and it showed a pH stability in the range of pH 4.0 to 12.0.

• Journal of Life Science
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• v.19 no.9
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• pp.1218-1225
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• 2009

Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.56-63
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• 2013

We have characterized the putative ${\alpha}$-glucosidase gene (st2525) selected by total genome analysis from the acidothermophilic crenarchaeon Sulfolobus tokodaii strain 7. The ORF was cloned and expressed as a fusion protein in Escherichia coli, and recombinant ST2525 was purified by Ni-NTA affinity chromatography. Maximum activity was observed at $95^{\circ}C$ and pH 4.0, and the enzyme exhibited stability with half-lives of 40.1 min and 7.75 min at extremely high temperatures of $100^{\circ}C$ and $105^{\circ}C$, respectively. The enzyme retained at least 85% of its maximal activity in the pH range of 4.0-11.0. ST2525 exclusively hydrolyzed ${\alpha}$-1,4-glycosidic linkages of oligosaccharides in an exo-type manner, with highest catalytic efficiency toward maltotriose. The enzyme also displayed transglycosylation activity, converting maltose to isomaltose, panose, maltotriose, isomaltotriose, etc. From these results, ST2525 could be potentially useful for starch hydrolysis as well as novel synthesis of oligosaccharides in industry.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.4
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• pp.663-668
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• 2000

To develop natural intermediate flavoring substances, optimal processing conditions and qualities for two stage enzyme hydrolysate (TSEH) from low-utilized small longfinned squid were investigated. The optimal conditions for TSEH method were found as digestion with Alcalase (0.2% w/w-sample, pH 8.0) at 55$^{\circ}C$ 3 hours at the 1st stage and with Neutrase (exo-peptidase, 0.2% w/w-sample, pH 6.0) at 45$^{\circ}C$ for 2~3 hours at the 2nd stage. Among the method of water extract, autolytic extract and various kinds yields, transparency and organoleptic taste. From the results of chemical experiments and sensory evaluation, longfinned squid TSEH is flavorable as the natural intermediate taste-active substances for fisheries products such as soup base, squid-taste pasty and snacks.

• Journal of Animal Science and Technology
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• v.60 no.10
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• pp.24.1-24.8
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• 2018

Background: Following the ban on the importation of import-dependent fed ingredients in most developing countries, the need to look inward for local content is now compelling. Thus, leaf meals that have phytogenic additive potentials are envisaged will be a viable feed ingredient in rabbit diets. Methods: The effect of dietary inclusion of gliricidia leaf meal (GLM) with or without multi-enzyme (E) supplementation in rabbits was investigated using ninety-six 35-day old rabbits of crossbreed (Newzealand and Chinchilla). One basal diet that met the requirements of growing rabbit was formulated (Diet 1). Thereafter, another two diets were formulated to contain 15% GLM and 15% GLM plus multi-enzyme at 1 g/kg and designated as diets 2 and 3 respectively. The rabbits were randomly distributed into the 3 diets (32 rabbits/treatment; 4 rabbits/replicate) and fed their respective experimental diets for 8 weeks. Results: The body weight and daily weight gain of the rabbits fed on GLM free diet and those on GLM-based diets (diets 1 and 2) were similar at finishing period of 63-91 day but have lower (P < 0.01) values than those rabbits fed GLM + E based diet (diet 3) at finishing period (63-91 days) and whole fattening period (35-91 days). The apparent dry matter and crude protein digestibility of rabbits fed control diet and those fed 15% GLM based diet were lower (P < 0.05) than those fed 15% GLM + E-based diet. Triglycerides concentration of rabbits fed 15% GLM-based diet without enzyme addition were lower (P < 0.05) than those observed for rabbits on the rest test diets. Cholesterol and Low-Density Lipoprotein levels of rabbits fed 15% GLM and 15% GLM + E-based diets were lower (P < 0.05) than those fed the GLM free diet. The superoxide dismutase and glutathione peroxidase of rabbits fed the GLM free diet (diet 1) were significantly (P < 0.05) lower than those fed the 15%GLM and 15% GLM + E-based diets. Conclusion: Dietary inclusion of GLM at 15% of the diet did not have a negative effect on the rabbits postweaning period (35-63 days) but will require multi-enzyme supplementation to enhance growth indices at finishing period (63-91 day) without precipitating negative effect on the rabbits' health status.