• 제목/요약/키워드: Enzyme Conversion

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Using Enzyme Supplemented, Reduced Protein Diets to Decrease Nitrogen and Phosphorus Excretion of White Leghorn Hens

  • Jacob, Jacqueline P.;Ibrahim, Sami;Blair, Robert;Namkung, Hwan;Paik, In Kee
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권12호
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    • pp.1743-1749
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    • 2000
  • An experiment was conducted to investigate the effect of supplementation of commercial phytase and ${\beta}-glucanase$ to wheat-soybean meal based layer diets. Control (17% CP) and reduced protein (13.5% CP) diets were compared with and without phytase and/or ${\beta}-glucanase$. Reducing dietary crude protein levels reduced the amount of N excreted by laying hens with no adverse affect on egg production or overall feed conversion ratio. There was, however, a slight reduction in average egg weight. When phytase was added to the control protein diets it was possible to reduce the level of dicalcium phosphate in the diet without a loss in performance and daily P output was reduced significantly. When phytase was added to the reduced protein diets, however, there was a dramatic loss in performance in the last four weeks of the study. Supplementation of ${\beta}-glucanase$ to wheat based layer diet did not appear to have beneficial affects in terms of laying performance and reducing nitrogen or phosphorus excretion. Combination of phytase and ${\beta}-glucanase$ had no positive effects on laying performance or reduction of DM, N and P.

Characterization and Culture Optimization of Regiospecific Cyclosporin Hydroxylation in Rare Actinomycetes Species

  • PARK, NAM-SIL;MYEONG, JI-SEON;PARK, HYUN-JOO;HAN, KYU-BOEM;KIM, SANG-NYUN;KIM, EUNG-SOO
    • Journal of Microbiology and Biotechnology
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    • 제15권1호
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    • pp.188-191
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    • 2005
  • Abstract Cyclosporins are a family of clinically-important immunosuppressive cyclic peptides produced by Tolypocladium inflatum. The structural modification of cyclosporins via hydroxylation at various positions of N-methyl leucines in cyclosporin A leads to a dramatic change of their bioactive spectra. Among over 100 soil actinomycetes screened, two actinomycetes species, Sebekia benihana and Pseudonocardia autotrophica, were identified to contain superior cyclosporin A hydroxylation activities. A HPLC-based cyclosporin A hydroxylation assay revealed that each strain possesses distinctive hydroxylation specificity and regiospecificity; mono-hydroxylation at the 4th N-methyl leucine of cyclosporin A by S. benihana, and di-hydroxylations at both 4th and 9th N-methyl leucines of cyclosporin A by P. autotrophica. The conversion yields for cyclosporin A hydroxylation by both S. benihana and P. autotrophica were significantly improved from less than 10% and 18% up to 58% and 45%, respectively, in the optimized culture containing molybdenum with 0.05 g/l of cyclosporin A concentration. An ancymidol-specific inhibition of cyclosporin hydroxylation also suggested that the regiospecific cyclosporin hydroxylation might be catalyzed by a putative cytochrome P450 mono-oxygenase enzyme.

Dietary chitosan enhances hepatic CYP7A1 activity and reduces plasma and liver cholesterol concentrations in diet-induced hypercholesterolemia in rats

  • Moon, Min-Sun;Lee, Mak-Soon;Kim, Chong-Tai;Kim, Yang-Ha
    • Nutrition Research and Practice
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    • 제1권3호
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    • pp.175-179
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    • 2007
  • The present study was performed to elucidate the hypocholesterolemic action of chitosan on the diet-induced hypercholesterolemia in rats. Male Sprague-Dawley rats (n=24) were fed with chitosan-free diet (Control), diets containing 2% or 5% chitosan for 4 weeks. Hypercholesterolemia was induced by adding 1% cholesterol and 0.5% cholic acid to all diets. Body weight gain and food intake of rats did not differ among the groups. The chitosan treated groups showed significant improvement in the plasma concentration of total cholesterol and LDL-cholesterol compared to the control group (p<0.05). Also, the chitosan treated groups decreased the liver concentration of total lipid and total cholesterol compared to the control group (p<0.05). The activity of hepatic cholesterol $7{\alpha}-hydroxylase$ (CYP7A1), the rate-limiting enzyme in the conversion of cholesterol to bile acids, was increased by 123% and 165% for the 2% or 5% chitosan diets, respectively. These findings suggest that enhancement of hepatic CYP7A1 activity may be a mechanism, which can partially account for the hypocholesterolemic effect of dietary chitosan in cholesterol metabolism.

키토산 비드에 고정화된 셀룰라아제의 특성 (Properties of Cellulase Immobilized on Chitosan Beads)

  • 이상헌;하용일;김보영;김범수
    • KSBB Journal
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    • 제29권4호
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    • pp.239-243
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    • 2014
  • Recently, there is a growing interest in efficient biomass pretreatment and saccharification processes to produce biofuels and biochemicals from renewable non-food biomass resources. In this study, glucose was produced from cellulose by immobilizing cellulase enzyme on chitosan beads which was reported to have high pH and temperature stability. The immobilized amounts of cellulase on chitosan beads linearly increased with increasing the concentrations of cellulase solution. The glucose production increased to 7.2 g/L from 1% carboxymethyl cellulose (CMC) substrate when immobilized at 20% cellulase solution. The maximum specific activity was 0.37 unit/mg protein when immobilized at 8% cellulase solution. At pH 7 and $37^{\circ}C$, the optimum reaction composition was 0.5 g beads/L from 1% CMC substrate. At this condition, the conversion to glucose completed at ca. 20 min.

Biochemical Changes during Embryonic Diapause in Domestic Silkworm, Bombyx mori L. (Lepidoptera: Bombycidae)

  • Singh, Tribhuwan;Saratchandra, Beera
    • International Journal of Industrial Entomology and Biomaterials
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    • 제5권1호
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    • pp.1-12
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    • 2002
  • Ecophysiologically diapause represents a syndrome of physiological and biochemical characteristics, all of which ensure survival during a long period of dormancy. Since, silkworm enters diapause as embryo at the early embryonic stage, the duration of egg life depends on the duration of embryonic diapause. The nature of diapause in silkworm, Bombyx mori, is primarily determined by genetic characters and endocrinologicnl mechanisms, mediated by environmental factors such as temperature and photoperiod. Hibernating potency value besides nucleic acid and carbohydrate metabolism, production and utilization of sorbitol are also equally responsible for induction, initiation, determination, maintenance and termination of diapause. Embryonic diapause in Bombyx moir, induced by active secretion of sub-oesophageal ganglion is attributed to hormonal system and metabolic adjustment, which serves to bring about a new physiological state. Metabolic conversion of trehalose to glycogen at induction, glycogen to sorbitol at initiation and sorbitol to glycogen at termination of diapause is correlated and in each metabolic shift a key enzyme becomes active in response to hormonal and environmental stimulation. An attempt has been made in this review article to discuss briefly the nature of embryonic diapause, influence of various factors on diapause nature, hormonal mechanism of diapause besides biochemical composition of egg, nucleic acid and carbohydrate metabolism, production and utilization of sorbitol in relation to induction, determination, maintenance, initiation and termination of diapause in the silkworm, Bombyx mori.

Isolation and Characterization of Kasugamycin Biosynthetic Genes from Streptomyces kasugaensis KACC 20262

  • JO YOU-YOUNG;LIU JING;JIN YING-YU;YANG YOUNG-YELL;SUH JOO-WON
    • Journal of Microbiology and Biotechnology
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    • 제15권3호
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    • pp.491-496
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    • 2005
  • The biosynthetic gene cluster for the aminoglycoside antibiotic kasugamycin was isolated and characterized from the kasugamycin producing strain, Streptomyces kasugaensis KACC 20262. By screening a fosmid library using kasA, the gene encoding aminotransferase, we isolated a 22 kb DNA fragment. The fragment contained seventeen complete open reading frames (ORFs); one of these ORFs, kasD, was identified as the gene for dNDP-glucose 4,6-dehydratase, which catalyzes the conversion of dNDP-glucose to 4-keto-6-deoxy-dNDP-glucose. The enzyme showed a broad spectrum of substrate specificity. In addition, ksR was overexpressed in E. coli BL21 and proved to be a self-resistance gene against kasugamycin. These findings suggest that the isolated gene cluster is highly likely responsible for the biosynthesis of kasugamycin.

Cloning and Characterization of a Glyoxalase I Gene from the Osmotolerant Yeast Candida magnoliae

  • Park, Eun-Hee;Lee, Dae-Hee;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • 제21권3호
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    • pp.277-283
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    • 2011
  • Glyoxalase I catalyzes the conversion of methylglyoxal to S-D-lactoylglutathione in the presence of glutathione. The structural gene of glyoxalase I (GLO1) was cloned from an osmotolerant yeast, Candida magnoliae, which produces a functional sweetener, erythritol, from sucrose. DNA sequence analysis revealed that the uninterrupted open reading frame (ORF) of C. magnoliae GLO1 (CmGLO1) spans 945 bp, corresponding to 315 amino acid residues, and shares 45.2% amino acid sequence identity to Saccharomyces cerevisiae Glo1. The cloned ORF in a multicopy constitutive expression plasmid complemented the glo1 mutation of S. cerevisiae, confirming that it encodes Glo1 in C. magnoliae. The responses of CmGLO1 to environmental stresses were different from those of S. cerevisiae, which only responds to osmotic stress. An enzyme activity assay and reverse transcription polymerase chain reaction revealed that the expression of CmGLO1 is induced by stress inducers such as methylglyoxal, $H_2O_2$, KCl, and NaCl. The GenBank Accession No. for CmGLO1 is HM000001.

Expression and purification of human mPGES-1 in E. coli and identification of inhibitory compounds from a drug-library

  • Kim, Woo-Il;Choi, Kyung-A;Do , Hyun-Soo;Yu, Yeon-Gyu
    • BMB Reports
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    • 제41권11호
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    • pp.808-813
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    • 2008
  • Human microsomal prostaglandin E synthase-1 (mPGES-1) is a membrane associated protein that catalyzes the conversion of prostaglandin $H_2$ ($PGH_2$) into prostaglandin $E_2$ ($PGE_2$). In this study, the expression of human mPGES-1 in E. coli was significantly enhanced by modifying the utility of specific codons and the recombinant mPGES-1 was efficiently purified to homogeneity. The $K_m$ and $V_{max}$ of the purified enzyme were determined and the trimeric state characterized by chemical cross-linking with glutaraldehyde. The purified mPGES-1 was used for the screening of a chemical library of bioactive or drug compounds to identify novel inhibitors, and oxacillin and dyphylline were identified as moderately inhibiting mPGES-1 with $I_{C50}$ values of 100 and 200 ${\mu}M$, respectively. As these compounds competitively inhibited the catalysis of $PGH_2$, their binding sites appeared to be located near the $PGH_2$ binding pocket.

Optimization of Ascorbic Acid-2-Phosphate Production from Ascorbic Acid Using Resting Cell of Brevundimonas diminuta

  • Shin, Woo-Jung;Kim, Byung-Yong;Bang, Won-Gi
    • Journal of Microbiology and Biotechnology
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    • 제17권5호
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    • pp.769-773
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    • 2007
  • With the aim to produce ascorbic acid-2-phosphate(AsA-2-P) from L-ascorbic acid(AsA, Vitamin C), nine bacteria conferring the ability to transform AsA to AsA-2-P were isolated from soil samples alongside known strains from culture collections. Most isolates were classified to the genus Brevundimonas by 16S phylogenetic analysis. Among them, Brevundimonas diminuta KACC 10306 was selected as the experimental strain because of its the highest productivity of AsA-2-P. The optimum set of conditions for the AsA-2-P production from AsA using resting cells as the source of the enzyme was also investigated. The optimum cultivation time was 16 h and the cell concentration was 120g/l(wet weight). The optimum concentrations of AsA and pyrophosphate were 550mM and 450mM, respectively. The most effective buffer was 50mM sodium formate. The optimum pH was 4.5 and temperature was $40^{\circ}C$. Under the above conditions, 27.5g/l of AsA-2-P was produced from AsA after 36 h of incubation, which corresponded to a 19.7% conversion efficiency based on the initial concentration of AsA.

담배 (Nicotiana tabacum L.) 현탁배양세포의 Elicitor 유도성 5-epi-Aristolochene Hydroxylase (Elicitor-Inducible 5-epi-Aristolochene Hydroxylase in Suspension Cultures of Tobacco (Nicotiana tabacum L.))

  • KWON, Soon-Tae;CHAPPELL, Joseph
    • 식물조직배양학회지
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    • 제25권3호
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    • pp.141-146
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    • 1998
  • [$^3$H] 5-epi-aristolochene (5-EAS)를 담배 현탁배양세포에 투여하여 elicitor에 의해 유도된 세포가 생합성하여 배지로 방출하는 [$^3$H]-capsidiol의 량을 측정함으로써 5-EAS hydroxylase의 활성을 검정하였고, 이 반응의 전 단계에 관여하는 효소인 sesquiterpene cyclase의 발현특성과 비교하였다. 5-EAS hydroxylase는 정상세포에는 전혀 활성을 보이지 않으나 elicitor로써 cellulase를 처리한 세포는 9시간 후부터 유도를 시작하여 18시간 후에 최대 활성을 보였는데 동일한 세포내에서 유도되는 cyclase와 유사한 패턴을 보였다. Cyt P450계 효소의 특이적 억제제로 알려진 ancymidol과 ketoconazole에 의해 5-EAS hydroxylase의 활성은 강한 억제를 보인 반면 cyclase의 활성은 억제를 보이지 않아 5-EAS hydroxylase가 P450계 효소임이 시사되었다.

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