• Title/Summary/Keyword: 박상동

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Enzymatic Properties of Barley $\alpha$-Amylase Chimeric Enzymes Produced by Staggered Extension Process (Staggered Extension Process를 통해 제조한 보리 알파아밀라제 Chimera 효소의 특성)

  • Kim, Tae-Jip;Choi, Seung-Ho;Jang, Myoung-Uoon;Park, Jung-Mi;Svensson, Birte
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.151-157
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    • 2010
  • Barley malt produces two different $\alpha$-amylase isozymes (AMY1 and AMY2), which share up to 80% of amino acid sequence identity with each other. However, their enzymatic properties differ remarkably. In this study, five chimeric enzymes between AMY1 and 2 were constructed by staggered extension process (StEP) technique, and their enzymatic properties were characterized. According to the results, chimeric AMY-D2, D8, and E12 showed the mixed or intermediate types of calcium-dependent activity between AMY1 and 2. Meanwhile, only AMY-E10 chimera could be significantly inhibited by barley $\alpha$-amylase/subtilisin inhibitor (BASI) protein. Chimera AMY-C6 showed the same calcium-dependency as AMY1, while AMY-E10 was closely similar to AMY2. As a result, it can be proposed that some amino acid residues in the region II, III, and IV of barley $\alpha$-amylases can play very important roles in the interaction with BASI, and those in III, V, VI, and VII may partly affect on the calcium-dependent activity.

S-adenosyl-L-homocysteine hydrolase gene is down-regulated in abnormal flower inducing environment in chyrsanthemum (국화 기형화 발생과 S-adenosyl-L-homocysteine hydrolase 유전자 발현)

  • Huh, Yeun Joo;Park, Sang Kun;Lim, Jin Hee;Choi, Seong Youl;Lee, Young Ryan
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.4
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    • pp.278-283
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    • 2010
  • This study was conducted to investigate the involvement of flower abnormality and S-adenosyl-L-homocysteine hydrolase (SAHH), which is one of the key enzyme in the maintenance of methylation. Plants exposed to high temperature (HT) and long day (LD) condition from 14-27days after short day (SD) produced abnormal flower, having numbers of ray florets. Numbers of ray florets were increased more than 2 folds by HT of $35/20^{\circ}C$ and LD of 14 hour comparing those of $25/20^{\circ}C$ (12 h/12 h). Full-length cDNA clone of S-adenosyl-L-homocysteine hydrolase (DgSAHH) in spray chrysanthemum 'Lerbin' contained an 1455 bp open reading frame coding for 485 amino acids. It showed highly conserved coding sequences among the different plant species with over 90% homology. DgSAHH expression was decreased in abnormal flower inducing treatment of HT and LD, while DgSAHH transcripts accumulated in flower bud of non abnormality inducing condition. This result implicate that DgSAHH expression is affected by temperature and photoperiod during flower development and suppression of DgSAHH is a one of the cause of abnormal flower under HT and LD condition.

Genetic relatedness of white spot syndrome virus (WSSV) from imported frozen shrimp (수입 냉동새우에서 검출된 WSSV의 유전학적 근연관계 조사)

  • Choi, So Won;Baek, Eun Jin;Choi, Ji Yeong;Tae, Won Jun;Kim, Hyoung Soon;Park, Woo Seong;Kim, Min Jae;Kim, Kwang Il
    • Journal of fish pathology
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    • v.34 no.2
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    • pp.141-147
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    • 2021
  • In this study, of the imported shrimps between 2017 and 2020, we investigated white spot syndrome virus (WSSV), covert mortality nodavirus (CMNV) and decapod iridescent virus 1 (DIV-1). Of the imported shrimps (a total of 29 groups), WSSV was detected as 31% (9/29) by nested PCR assay. And CMNV and DIV-1 were not identified in this study. To investigate the genetic relatedness of WSSV identified from imported shrimp, VR 14/15 region showed WSSV genomic variable loci was compared with reference isolates. Among the nine WSSV-positive samples, VR 14/15 region was amplified in only a sample (20-CH-1 isolate, imported from China in 2020). And the 20-CH-1 isolate showed 99.8% identity with WSSV-IN-05-01 which was reported in India in 2005, suggesting that those of WSSV have been spread from India to China. Furthermore, although the pathogenicity of WSSV identified from frozen shrimp was not evaluated, the international trade of diseased frozen shrimps could be led to the potential risk of virus transmission.

Protease Activity of Lactic Acid Bacteria Isolated from Korean Traditional Fermented Food (전통 발효식품으로부터 Protease 활성을 보유한 유산균의 분리 및 동정)

  • Kook, Moo Chang;Cho, Seok Cheol;Park, Hoon;Kim, Seung Seop;Pyun, Yu Ryang;Choi, Woon Yong;Lee, Hyeon Yong
    • Food Engineering Progress
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    • v.15 no.2
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    • pp.182-187
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    • 2011
  • A proteolytic lactic acid bacterium was isolated from Korean traditional fermented foods. The isolate BV-26, which had a protease activity (24 U/mg-crude protein), was identified as Lactobacillus plantarum by the API 50CHL kit and 16S rDNA analysis (99.9% of homology), and named as L. plantarum BV-26. Cell growth and protease activity of L. plantarum BV-26 was determined in MRS broth using 5L jar fermentor at $30^{\circ}C$. The maximum growth of L. plantarum BV-26 was reached at 18 hr in MRS broth, while protease activity of BV-26 was detectable at 12 hr and the highest activity was obtained after 16 hr cultivation. Therefore, we expect that the proteolytic lactic acid bacteria, L. plantarum BV-26, may be used as a starter for the fermentation of animal feed. Especially, the fermentation of soybean meal with the strain can be applied for improving feed utilization.

Occurrence of Leaf Spot Disease on Watermelon Caused by Pseudomonas syringae pv. syringae (Pseudomonas syringae pv. syringae에 의한 수박 잎점무늬병의 발생)

  • Park, Kyoung-Soo;Lee, Ji-Hye;Kim, Young-Tak;Kim, Hye-Seong;Lee, June-woo;Lee, Hyun-Su;Lee, Hyok-In;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.27 no.4
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    • pp.180-186
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    • 2021
  • Typical bacterial symptoms, water-soaking brown and black leaf spots with yellow halo, were observed on watermelon seedlings in nursery and field of Gyeongnam and Jeonnam provinces. Bacterial isolates from the lesion showed strong pathogenicity on watermelon and zucchini. One of them was rod-shaped with 4 polar flagella by observation of transmission electron microscopy. They belonged to LOPAT group 1. The phylogenical trees with nucleotide sequences of 16S rRNA and multi-locus sequencing typing with the 4 house-keeping genes (gapA, gltA, gyrB, and rpoD) of the isolates showed they were highly homologous to Pseudomonas syringae pv. syringae and grouped together with them, indicating that they were appeared as P. syringae genomospecies group 1. Morphological, physiological, and genetical characteristics of the isolates suggested they are P. syringae pv. syringae. We believe this is the first report that P. syringae pv. syringae caused leaf spot disease on watermelon in the Republic of Korea.

Effect of Cervi Pantotrichum Cornu Herbal acupuncture on protease activities, antioxidant in Rheumatoid arthritis rats (류마티스 관절염 실험용쥐의 활액에서 단백분해효소의 활성 및 항산화에 대한 녹용약침의 효과)

  • Park, Sang-Dong;Kim, Min-Jeong;Lee, A-Ram;Jang, Jun-Hyouk;Kim, Kyung-Ho
    • Journal of Acupuncture Research
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    • v.19 no.2
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    • pp.51-64
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    • 2002
  • We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.

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Egg Development and Larvae of the Rose Bitterling Rhodeus ocellatus (KNER) (흰줄납줄개의 난발생과 부화자어)

  • KIM Yong Uk;PARK Yang Seong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.6
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    • pp.586-593
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    • 1985
  • The rose bitterling, Rhodeus ocellatus (KNER) is commonly distributed in the fresh waters of Korea and Japan. On January 15, February 15, and March 18 in 1984, mature adults of rose bitterling were caught in the watercourse of Maeri, Kimhae, South Korea. The authors fertilized the eggs employing dry method in the laboratory on May 7, 16 and 25 in 1984. Hatched larvae were reared in small aquariums at $17{\sim}25.5^{\circ}C$ (average around $21.2^{\circ}C$). Mollusks, Anodonta woodiana in the gill chamber of which, rose bitterling lay eggs were caught in order to study natural spawning of the rose bitterling in the same watercourse. The eggs of this species are not adhesive and demersal. The size of the eggs varies from 2.54 to 2.75mm in long diameter and 1.45 to 1.65mm in short diameter. The eggs are cylindrical in form when they are extruded from ovipositor, immediately after entering water, but they acquire their distinctive form of a greatly elongated pear. Hatching took place in ca. 39 hours after fertilization. The newly hatched larvae were $2.65{\sim}2.70mm$ in total length possessing yolk sac and 13-14 myotomes. Thirteen days after hatching, the prelarvae attained 6.5 mm in total length, and the first melanophores appeared on the head, and the anterior part and sides of the yolk sac. One month after hatching, the postlarvae attained 8.5mm. in total length and emerged from the gills of the mollusks. Then the yolk sac was completely resorbed. Two months after hatching, the rose bitterling attained 14.4mm in total length, and entered the fingerling period of life. All the rays already present were the D. III, 11-12, A. III, 11-12, P. 10, V. 7. and a distinguishing, feature is the presence of a black pigment spot in the lobe of the dorsal fin.

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Identification of a new marine bacterium Ruegeria sp. 50C-3 isolated from seawater of Uljin in Korea and production of thermostable enzymes (대한민국 울진 연안 해양에서 분리한 해양 미생물 Ruegeria sp. 50C-3의 동정 및 내열성 효소 생산)

  • Chi, Won-Jae;Kim, Jong-Hee;Park, Jae-Seon;Hong, Soon-Kwang
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.344-351
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    • 2016
  • A marine bacterium, designated as strain 50C-3, was isolated from a seawater sample collected from the East Sea of South Korea. The strain is a Gram-negative, aerobic, yellow colored polar-flagellated bacterium that grows at $20-50^{\circ}C$ and pH 5.5-8.5. Optimal growth occurred at $40-50^{\circ}C$, at pH 6.5-7.5, and in the presence of 2% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, the isolate was considered to represent a member of the genus Ruegeria. The result of this analysis showed that strain 50C-3 shared 99.4% and 96.98% sequence similarity with Ruegeria intermedia CC-GIMAT-$2^T$ and Ruegeria lacuscaerulensis ITI-$1157^T$, respectively. Furthermore, strain 50C-3 showed clear differences from related strains in terms of several characteristics such as motility, carbon utilization, enzyme production, etc. The DNA G+C content was 66.7 mol%. Chemotaxonomic analysis indicated ubiquinone-10 (Q-10) as the predominant respiratory quinone. Based on phenotypic, chemotaxonomic, and phylogenetic characteristics, the isolate represents a novel variant of the Ruegeria intermedia CC-GIMAT-$2^T$, for which we named Ruegeria sp. 50C-3 (KCTC23890=DSM25519). Strain 50C-3 did not produce cellulase and agarase, but produced alkaline phosphatase, ${\alpha}$-galactosidase, and ${\beta}$-galactosidase. The three enzymes showed stable activities even at $50^{\circ}C$ and thus regarded as thermostable enzymes. Especially, the ${\beta}$-galactosidase activity enhanced by 1.9 times at $50^{\circ}C$ than that at $37^{\circ}C$, which may be very useful for industrial application.

Mutational Analysis of MECP2 Gene in 34 Rett Syndrome (Rett 증후군 34례의 MECP2 유전자 변이에 관한 연구)

  • Park, Sang Jo;Hwang, Tae Gyu;Son, Byeong Hee;Kim, Chul Min
    • Clinical and Experimental Pediatrics
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    • v.45 no.10
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    • pp.1263-1272
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    • 2002
  • Purpose : Rett syndrome(RTT) is an X-linked dominant neurodevelopmental disorder affecting 1 per 10,000-15,000 female births worldwide. It was initially described by Andreas Rett in 1966. RTT involves developmental regression characterized stereotypic hand movements, tremors, gait apraxia, seizures, deceleration of head growth after the age of 6-18 months. The disease-causing gene was identified as MECP2 on chromosome Xq28. We carried out mutational analysis of MECP2 genes in RTT patients. Methods : Whole blood(5 cc) of 34 sporadic RTT patients was collected in EDTA-anticoagulated tubes. Genomic DNA was extracted from peripheral blood using the E.Z.N.A. blood DNA kit. Four exons of the MECP2 gene were amplified by PCR in 34 Korean with RTT. We carried out PCR divided the exon three into two parts and the exon four into five parts. Primer sequences designed by Amir et al. in 1999 were almost used(AF030876). Sequencing primers used were the same as PCR. DNA sequencing reactions were performed using an ABI 377 DNA sequencer and ABI PRISM dye terminator cycle sequencing reaction kit(Perkin-elmer). The results were compared with the normal DNA sequence(X99686). To confirm the change of sequence on novel mutations, RFLP analysis was performed. Results : The MECP2 mutations were detected in 23(67.6%) of the 34 patients. The mutations consisted of 12 different types including nine missense and three nonsense mutations. Of these, three (L100V, G161E and T311M) mutations were newly identified. Most of the mutations discovered are located within MBD(39.1%) and TRD(39.1%). In this study, three(T158M, R270X, R306C) mutations were identified high frequency. Conclusion : MECP2 gene was also an important cause of Korean RTT patients. MECP2 gene study is an important tool for diagnosis of Korean RTT patients.

Characterization of an Extracellular Xylanase from Bacillus sp. HY-20, a Bacterium in the Gut of Apis mellifera (꿀벌(Apis mellifera)의 장내 세균인 Bacillus sp. HY-20이 분비하는 Xylanase의 특성)

  • Lee, Lan-Hee;Kim, Do-Young;Han, Mi-Kyoung;Oh, Hyun-Woo;Ham, Su-Jin;Park, Doo-Sang;Bae, Kyung-Sook;Sok, Dai-Eun;Shin, Dong-Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.332-338
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    • 2009
  • A xylan-decomposing bacterium, HY-20, was isolated from the gut of a honeybee, Apis mellifera, and identified as Bacillus sp. The extracellular GH11 xylanase (XylP) gene (687-bp) of strain HY-20 encoded a protein of 228 amino acids with a deduced molecular mass of 25,522 Da and a calculated pI of 9.33. The primary structure of XylP was 97% identical to that of B. pumilus xylanase (GenBank accession no.: AY526092) that has not been characterized yet. The recombinant His-tagged enzyme (rXylP) overexpressed in Escherichia coli BL21 harboring pET-28a(+)/xylP was purified to electrophoretic homogeneity by cation exchange and gel permeation chromatographies. The purified enzyme exhibited the highest catalytic activity toward birchwood xylan at pH 6.5 and $50^{\circ}C$ and retained approximately 50% of its original activity when pre-incubated at $55^{\circ}C$ for 15 min. The recombinant enzyme was completely inactivated by $Hg^{2+}$ (1 mM) and N-bromosuccinimide (5 mM), while its activity was slightly stimulated by approximately 10% in the presence of $Mn^{2+}$ (1 mM), $Fe^{2+}$ (1 mM), and sodium azide (5 mM). rXylP was able to efficiently degrade various polymeric xylose-based substrates but PNP-sugar derivatives and glucose-based polymers were not susceptible to the enzyme.