• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.552-559
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• 2008

Ethyl (S)-4-chloro-3-hydroxybutyrate is an intermediate for the synthesis of Atorvastatin, a chiral drug used for hypercholesterolemia. A Rhodococcus erythropolisstrain (No.7) able to convert 4-chloro-3-hydroxybutyronitrile into 4-chloro-3-hydroxybutyric acid has recently been isolated from soil. This activity has been regarded as having been caused by the successive actions of the nitrile hydratase and amidase. In this instance, the corresponding amidase gene was cloned from the R. erythropolis strain and expressed in Escherichia coli cells. A soluble active form of amidase enzyme was obtained at $18^{\circ}C$. The Ni column-purified recombinant amidase was found to have a specific activity of 3.89 U/mg toward the substrate isobutyramide. The amidase was found to exhibit a higher degree of activity when used with mid-chain substrates than with short-chain ones. Put differently, amongst the various amides tested, isobutyramide and butyramide were found to be hydrolyzed the most rapidly. In addition to amidase activity, the enzyme was found to exhibit acyltransferase activity when hydroxyl amine was present. This dual activity has also been observed in other enzymes belonging to the same amidase group (E.C. 3.5.1.4). Moreover, the purified enzyme was proven to be able to enantioselectively hydrolyze 4-chloro-3-hydroxybutyramide into the corresponding acid. The e.e. value was measured to be 52% when the conversion yield was 57%. Although this e.e. value is low for direct commercial use, molecular evolution could eventually result in this amidase being used as a biocatalyst for the production of ethyl (S)-4-chloro-3-hydroxybutyrate.

• Microbiology and Biotechnology Letters
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• v.18 no.5
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• pp.476-483
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• 1990

In order to investigate hydantoinase-producing strain of the genus Streptomyces, 523 strains of Streptomycee sp. isolated from soils were cultivated in various media and conversion activity of the enzyme was measured to DL-5-phenylhydantoin. A number of strains producing hydantoinase were detected and among them, the strain of Streptomyces sp. Y-183 was selected as a most powerful strain to producing the enzyme. The optimal culture conditions for the production of hydantoinase of the strain were studied, and it was found that almost all hydantoinase activity was produced in the cell fraction. The maximum activity of the enzyme, 17.8 unitstg of dried cells weight, was obtained when the strain was cultured at $30^{\circ}C$ for 72 hr in a medium containing 1.0% of glycerol. 0.5% of yeast extract. 0.5% of soytone, 0.5% of beef extract, 0.6% of KCI, 0.002% of $K_2HP0_4, 0.25% \;of \;CaC0_3, \;0.0002% \; of \; ZnSO_4, \; 0.0002%\; of\; FeS0_4$, and 0.4% of uracil as an inducer, and the pH of culture broth was adjusted ranging from 7.0 to 7.5.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.67-67
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• 2003

Tributyltin chloride (TBTCI) is an organotin compounds that have been widely used as antifouling agents and bioaccumulated in the food chain. TBTCI has been known to induce imposex in female gastropods. There are several reports that TBTCI increased testosterone level and inhibited the conversion of testosterone to estradiol by the aromatase cytochrome P450 enzyme. In this studies, we investigated the effects of TBTCI on steroidogenesis in testes, We dosed to 4-week-old Spragus-Dawleys (SD) male rats with TSTCI (0, 1, 5, 10, and 20mg/kg/day) daily by gavage for 14 days. TBTCI significantly decreased the weights of seminal vesicle, prostate, cowper's gland and LABC at 10 and 20mg/kg/day but significantly Increased the weights of liver at 10 and 20mg/kg/day and adrenals at 20mg/kg/day. mRNA levels of steroidogenic acute regulatory (StAR) and P450 aromatase were decreased and mRNA levels of cytochrome P450 17$\alpha$-hydroxylase/$C_{17-20}$ lyase (P450c17) were increased by TBTCI. TBTCI significantly increased serum testosterone level in dose-dependent manner. From above results, we found that TBTCI altered mRNA levels of enzymes related steroidogenesis, weights of organs and serum testosterone levels. This suggests that change of hormone levels may be due to alteration of mRNA levels of steroidogenic enzyme in testes, but further studies are necessary to investigate hormone levels in testis organ in order to find a relation of enzyme related to steroidogenesis with hormone levels. This work was supported by the Korea FDA Grant KFDA-03131-EDS-010.

• Applied Biological Chemistry
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• v.27 no.2
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• pp.112-118
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• 1984

Inverase was entrapped in calcium alginate gel. The immobilized beads had excellent uniform size and configuration. To screen the optimal conditions possessing high enzyme activity, effects of sodium alginate concentration, $CaCl_2$ concentration, and the incubation time of beads in $CaCl_2$ solution during the immobilization procedure were investigated. Immobilized beads prepared from the optimal conditions had 18.8 units per ml of gel, which is equivalent to 68% of the activity of soluble enzyme. Several kinetic parameters were determined: Km value. 143 mM; optimum pH, 4.5; optimum temperature, $50^{\circ}C$. Enzyme loading capacity was 150 mg per 20 ml gel. No significant decrease in sugar conversion was observed during the column operation for 6 days.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.299-305
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• 2006

Aromatic L-amino acid transaminase is an enzyme that is able to transfer the amino group from L-glutamate to unnatural aromatic ${\alpha}-keto$ acids to generate ${\alpha}-ketoglutarate$ and unnatural aromatic L-amino acids, respectively. Enrichment culture was used to isolate thermophilic Bacillus sp. T30 expressing this enzyme for use in the synthesis of unnatural L-amino acids. The asymmetric syntheses of L-homophenylalanine and L-phenylglycine resulted in conversion yields of >95% and >93% from 150 mM 2-oxo-4-phenylbutyrate and phenylglyoxylate, respectively, using L-glutamate as an amino donor at $60^{\circ}C$. Synthesized L-homophenylalanine and L-phenylglycine were optically pure (>99% enantiomeric excess) and continuously pre-cipitated in the reaction solution due to their low solubility at the given reaction pH. While the solubility of the ${\alpha}-keto$ acid substrates is dependent on temperature, the solubility of the unnatural L-amino acid products is dependent on the reaction pH. As the solubility difference between substrate and product at the given reaction pH is therefore larger at higher temperature, the thermophilic transaminase was successfully used to shift the reaction equilibrium toward rapid product formation.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2004.11a
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• pp.95-96
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• 2004

An experiment was conducted to investigate the effects of dietary supplementation of feather meal (FM its) digests on the performance of broiler chicks and taurine content in broiler meat. A total of 1,000 broiler chickens were assigned to five dietary treatments : Control, FM diet(FM), FM+pyridoxine(FM+Pyridox), H$_2$O$_2$ treated FM diet(H$_2$O$_2$-FM) and enzyme treated FM diet (Enzyme-FM). Treated diets were supplemented with FM or FM digests at the level of 5 % to the control diet. During the stater period, weight gain of chicks fed FM+Pyridox was significantly higher(P<0.05) than those of the other FM or FM digest treatments but was not different from the control. Weight gam of overall period were not significantly different among treatments. Feed intake of the control was greater than that of FM or FM digest treatments. Feed conversion ratio(feed intake/gain) of chicks fed FM and H$_2$O$_2$-FM were significantly higher than those of Enzyme-FM and FM+Pyridox, but were not significantly different from the control. Taurine contents of leg and breast mucle were significantly (P<0.01) different among treatments but those of liver were not significantly different. Taurine content of FM+Pyridox was highest in both leg and breast muscle. It was 85 % higher in leg muscle and 15 % higher in breast muscle than that of the control. Sensory evaluation data showed significant but not consistant responses in various parameters. FM + Pyridox treatment showed highest score in aroma of raw leg muscle of male and in juiciness and tenderness of broiled breast muscle of male chickens. Control group showed highest color score in raw leg muscle of female and lowest overall acceptability score in broiled breast and leg muscle of male chicken. It is concluded that taurine can be enriched especially in broiler leg meat by 5 % FM diet supplemented with pyridoxine.

• Journal of Chest Surgery
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• v.21 no.5
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• pp.803-815
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• 1988

The present experiments were performed to confirm the hypothesis that xanthine oxidase[XOD], as a source and mechanism of oxygen radical production, plays an important role in the genesis of the reperfusion injury of ischemic myocardium. The experimental ischemic-reperfusion injury was induced in isolated, Langendorff preparations of rat hearts by 60 min. Of global ischemia with aortic clamping followed by 20 min. of reperfusion with oxygenated Krebs-Henseleit solution[pH 7.4, 37*C]. The results were as follows: 1. The releases of creatine phosphokinase and a lipid peroxidation product, malondialdehyde[MDA] into the coronary effluent were abruptly increased upon reperfusion of ischemic hearts. The increases of the enzyme and MDA were suppressed significantly in the hearts removed from rats pretreated with allopurinol, a specific XOD inhibitor[20mg/kg, oral, 24 hrs and 2 hrs before study]. This effect of allopurinol was comparable to that of oxygen radical scavengers, superoxide dismutase[5, 000U] and catalase[12, 500 U]. 2. The increased SOD-inhibitable reduction of ferricytochrome C, which was infused to the hearts starting with reperfusion, was significantly suppressed in allopurinol pretreated hearts. 3. Activities of myocardial XOD were compared in the normal control hearts and the ischemic ones. Total enzyme activities were not different in both hearts. However, comparing with the control, the ischemic ones showed higher activity in 0-form and lower activities in D-form and D/O-form. 4. In the ischemic hearts, phenylmethylsulfonyl fluoride, a serine protease inhibitor, prevented significantly the increase of 0-form and the decreases of D and D/O-form, while thiol reagents did not affect the changes of the enzyme. 5. The increase of 0-form and the decreases of D and D/0-form were not significant in both calcium-free perfused and pimozide, a calmodulin inhibitor, treated ischemic hearts. 6. The SOD-inhibitable reduction of ferricytochrome C were suppressed by PMSF and pimozide treatment as well as by calcium-free perfusion. It is suggested from these results that in the ischemic rat myocardium, xanthine oxidase is converted to oxygen radical producing 0-form by calcium, calmodulin-dependent proteolysis and plays a contributing role in the genesis of ischemic-reperfusion injury by producing oxygen free radicals.

• Microbiology and Biotechnology Letters
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• v.2 no.1
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• pp.45-50
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• 1974

1 With cysteinedesulfhydrase (E. C.4.4.1.1.) from Aerobactor aerogenes, an enzyme which catalyzes the stoichiometric conversion of L-cysteine to pyruvate, ammonia and sulfide, reversibility of the degradation of L-cysteine was investigated. It was found that the enzyme also catalized the reverse reaction of $\alpha$, $\beta$-elimination to synthesize L-cysteine derivatives from pyruvate, ammonia and sulfides when large amounts of substrates were added to the reaction mixtures. 2. The synthetic reaction by cysteinedesulfhydrase proceeded linearly with incubation time and enzyme concentrations. The optimal pH for the synthetic reaction was 10.0. 3. The results of the isolation and identification of the products showed that the L-cysteine derivatives synthesized by this enzymatic method were identical with S-methyl-L-cysteine and S-ethyl-L-cysteine respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.2
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• pp.181-190
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• 2020

This study was conducted to estimate the optimum dietary DHA oil level and replacement level of enzyme treated fish meal (EFM) with sardine fish meal for juvenile Atlantic bluefin tuna Thunnus thynnus. Four diets were used: 1) EFM75 in which 75% EFM and 4% DHA oil were applied, 2) EFM60, with 60% EFM and 15% sardine fish meal, 3) DHA2 with 2% of DHA oil, and 4) SL as a raw fish feed. In a feeding trial, juvenile bluefin tuna (body weight 30.1 g) were randomly stocked into four experimental tanks (69 tones) and fed the experimental diets for 13 days. Fish weight gain was higher in the EFM75 and SL groups than in the DHA2 and EFM60 groups. The feed conversion ratio was lower in the EFM75 and DHA2 groups than in the EFM60 and SL groups. Survival was higher in fish fed the formulated diet groups (EFM75, EFM60 and DHA2) than in fish fed SL. This study clearly indicates that up to 10% dietary sardine fish meal can be used in juvenile T. thynnus diets, with an optimum dietary DHA oil level of approximately 3%.

• Archives of Pharmacal Research
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• v.29 no.3
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• pp.241-248
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• 2006

Angiotensin converting enzyme (ACE) inhibitors have cardioprotective effects in different species including human. This cardioprotective effect is mainly due to the inhibition of bradykinin (BK) degradation rather than inhibition of the conversion of angiotensin I to angiotensir. II. Bradykinin, a nonapeptide, has been considered to be the potential target for various enzymes including ACE, neutral endopeptidase 24.11, carboxypeptidase M, carboxypeptidase N, proline aminopeptidase, endopeptidase 24.15, and meprin. In the present study, the coronary vascular beds of Sprague Dawley rat isolated hearts were perfused (single passage) with Krebs solution alone or with different concentrations of BK i.e. $2.75{\times}10^{-10},\;10^{-7},\;10^{-6}\;and\;10^{-5}M$ solution. Percent degradation of BK was determined by radioimmunoassay. The degradation products of BK after passing through the isolated rat-hearts were determined using RP-HPLC and mass spectroscopy. All the four doses of BK significantly decreased the perfusion pressure during their passage through the hearts. The percentage degradation of all four doses was decreased as the concentration of drug was increased, implying saturation of a fixed number of active sites involved in BK degradation. Bradykinin during a single passage through the hearts degraded to give [1-7]-BK as the major metabolite, and [1-8]-BK as a minor metabolite, detected on HPLC. Mass spectroscopy not only confirmed the presence of these two metabolites but also detected traces of [1-5]-BK and arginine. These findings showed that primarily ACE is the major cardiac enzyme involved in the degradation of bradykinin during a single passage through the coronary vascular of bed the healthy rat heart, while carboxypeptidase M may have a minor role.