• 제목/요약/키워드: hybrid enzyme

검색결과 70건 처리시간 0.023초

Physiological Functionalities of Vitis hybrid (Sheridan)-Rubus coreanus Red Wine Made by Saccharomyces cerevisiae

  • Jang, Jeong-Hoon;Kim, Jae-Ho;Ahn, Byung-Hak;Lee, Jong-Soo
    • Mycobiology
    • /
    • 제39권2호
    • /
    • pp.109-112
    • /
    • 2011
  • Vitis hybrid (Sheridan)-Robus coreanus red wine was vinified by fermentation of a mixture of Vitis hybrid and Robus coreanus must at $25^{\circ}C$ for 10 days. The Vitis hybrid-Robus coreanus red wine had ethanol contents of 10.9%. It had high antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity of 57.8% and antioxidant activity of 64.8%. Changes in the physicochemical properties and functionality of the Vitis hybrid-Robus coreanus red wine was investigated during a post-fermentation period of three months. The ACE inhibitory activity of the red wine increased as the post-fermentation period prolonged, and showed the highest ACE inhibitory activity of 70.4% 60 days post-fermentation. However, the antioxidant activity declined significantly to 47.2% during the post-fermentation period of 60 days. In terms of sensory evaluation, the Vitis hybrid-Robus coreanus red wine had the best acceptability 60 days post-fermentation.

표고 교잡균주들의 세포외 laccase 활성 검출에 미치는 배지성상과 발색반응 시약의 영향 분석 (Analysis of the Effect of Media Types and Chromagenic Chemicals on the Detection of Extracellular Laccase Activity among Lentinula edodes Strains)

  • 김준영;권혁우;당룡장;고한규;김성환
    • 한국균학회지
    • /
    • 제39권1호
    • /
    • pp.48-52
    • /
    • 2011
  • 본 연구에서는 표고 교잡균주들의 생화학적 특성을 이해하기 위한 하나의 시도로서 교잡균주간의 laccase 효소의 활성을 비교하는 빠르고 간편한 방법을 모색하고자 균사체 배양 배지의 성상과 염색물질이 laccase 활성 검출에 미치는 영향을 조사 비교하였다. 고체배지와 액체배지 모두에서 발색시약이 Remazol Brilliant Blue R(RBBR)일 경우는 potato dextrose 성분 보다 malt extract 성분 배지를 사용하는 것이 표고버섯균 균사가 분비하는 세포외 laccase 활성 검출이 용이하였다. Guaiacol을 발색시약으로 사용하는 경우는 potato dextrose 성분과 malt extract 성분 배지 모두 세포외 laccase 활성 검출이 용이하였다. 2ml microfuge tube를 이용한 액체 배양 방법은 RBBR와 Guaiacol 모두 malt extract 성분배지에서 3일만에 표고 교잡균주간의 세포외 laccase 활성 검출과 정량적 비교를 가능케 하였다.

Polychlorinated Biphenyl에 의한 백서간 Cytochrome P-$450_{LMII}$에 대한 Monoclonal Antibody 생성에 관한 연구 (Production of Monoclonal Antibody to Polychlorinated Biphenyl Induced Cytochrome P-450 LMII in Rat Liver)

  • 김정희;김재룡;이기영
    • Journal of Yeungnam Medical Science
    • /
    • 제3권1호
    • /
    • pp.103-110
    • /
    • 1986
  • Polychlorinated biphenyl(PCB)에 유도된 rat liver cytochrome $P_{450LMII}$을 Balb/c mouse에 주사하여 면역된 spleen cells과 $SP_2$ myeloma cell을 polyethylene glycol(PEG 3500)으로 세포융합 시켜 얻은 fused cell을 계속 배양하여 cloning을 반복하고 ELISA로 확인하여 monoclonal antibody(Mab)를 생산하는 hybrid cell을 얻었으며 mouse 복강내에 hybrid cell(${\times}10^7$)을 주사하여 ascites를 모아 cellulose ion exchange chromatography로 Mab을 정제하였으며 $I^{125}$로 label 시킨 Mab는 $CP_{450LMII}$ 항원과 hybridization시켜 binding을 관찰하였으며 SDS-polyacrylamide 전기영동에서 분자량 55,000 및 110,000인 두 개의 band를 관찰하였다.

  • PDF

Biochemical Performance and Quantitative Assessment of F1 Hybrid of Two Ecoraces of Tropical Tasar Silkworm Antheraea Mylitta Drury (Lepidoptera: Saturniidae)

  • Lokesh, Gangadharaiah;Tirkey, Sushma Rani;Srivastava, Ashok Kumar;Kar, Prasant Kumar;Sinha, Manoj Kumar
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • 제26권2호
    • /
    • pp.67-73
    • /
    • 2013
  • Antheraea mylitta Drury is basically a crossbreeding species, as such it seems to be potentially a good material for the exploitation of heterosis. In the present study F1 hybrid of wild ecorace Laria (L) and semi-domestic Daba (D) was raised and evaluated for various quantitative traits and biochemical parameters during larval stage. Improved fecundity ($+18{\pm}1.8%$ and higher egg hatching rate ($+10.96{\pm}1.3%$) was recorded in the F1hybrid ($L{\times}D$). Biochemical parameters studied in the hemolymph, midgut and fatbody of the larva showed significantly higher (P<0.05) total proteins and carbohydrate concentration besides digestive enzyme activity. Correspondingly SDS-PAGE revealed more number of protein bands in the hemolymph sample of F1s, ranging between 29 kDa to 66 kDa compared to parental lines. The present study demonstrates the positive heterosis effect in the F1 hybrid of Laria ${\times}$ Daba. Biochemical analysis indicates that, there is possibilities of exploitation of hybrids with specific parents targeted for desirable commercial traits (silk yield and fecundity). Moreover, most of these biochemical parameters can be used as markers to analyze the genetic improvement in the tasar silkworms.

Organic-Inorganic Hybrid Nanoflowers as Potent Materials for Biosensing and Biocatalytic Applications

  • Tran, Tai Duc;Kim, Moon Il
    • BioChip Journal
    • /
    • 제12권4호
    • /
    • pp.268-279
    • /
    • 2018
  • Flower-shaped organic-inorganic hybrid nanostructures, termed nanoflowers, have received considerable recent attention as they possess greatly enhanced activity, stability, durability, and even selectivity of entrapped organic biomolecules, which are much better than those from the conventional methods. They can be synthesized simply via co-incubation of organic and inorganic components in aqueous buffer at room temperature and yield hierarchical nanostructures with large surface-to-volume ratios, allowing for low-cost production by easy scale-up, as well as the high loading capacity of biomolecules without severe mass transfer limitations. Since a pioneering study reported on hybrid nanoflowers prepared with protein and copper sulfate, many other organic and inorganic components, which endow nanoflowers with diverse functionalities, have been employed. Thanks to these features, they have been applied in a diverse range of areas, including biosensors and biocatalysis. To highlight the progress of research on organic-inorganic hybrid nanoflowers, this review discusses their synthetic methods and mechanisms, structural and biological characteristics, as well as recent representative applications. Current challenges and future directions toward the design and development of multi-functional nanoflowers for their widespread utilization in biotechnology are also discussed.

Molecular Cloning and Expression of a Xylanase Gene from Alkalophilic Bacillus sp.

  • Yu, Ju-Hyun;Kang, Yun-Sook;Park, Young-Seo;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
    • /
    • 제1권4호
    • /
    • pp.251-255
    • /
    • 1991
  • A 16 kilobase (kb) HindIII fragment of alkalophilic Bacillus sp. YC-335 containing a gene for xylanase synthesis was inserted at the HindIII site of pBR322 and cloned in Escherichia coli HB101. After subcloning of recombinant plasmid pYS52, the 1.5 kb fragment was found to code for xylanase activity, and the hybrid plasmid was named pYS55. The DNA insert of the plasmid was subjected to restriction enzyme mapping, which showed that pYS55 had single site for PuvII and SstI in the 1.5 kb insert fragment. Southern hybridization analysis revealed that the cloned gene was hybridized with chromosomal DNA from alkalophilic Bacillus sp. YC-335. About 64% of the enzyme activity was observed in the extracellular and periplasmic space of E. coli HB10l carrying pYS55.

  • PDF

Protein Analysis Using a Combination of an Online Monolithic Trypsin Immobilized Enzyme Reactor and Collisionally-Activated Dissociation/Electron Transfer Dissociation Dual Tandem Mass Spectrometry

  • Hwang, Hyo-Jin;Cho, Kun;Kim, Jin-Young;Kim, Young-Hwan;Oh, Han-Bin
    • Bulletin of the Korean Chemical Society
    • /
    • 제33권10호
    • /
    • pp.3233-3240
    • /
    • 2012
  • We demonstrated the combined applications of online protein digestion using trypsin immobilized enzyme reactor (IMER) and dual tandem mass spectrometry with collisionally activated dissociation (CAD) and electron transfer dissociation (ETD) for tryptic peptides eluted through the trypsin-IMER. For the trypsin-IMER, the organic and inorganic hybrid monolithic material was used. By employing the trypsin-IMER, the long digestion time could be saved with little or no sacrifice of the digestion efficiency, which was demonstrated for standard protein samples. For three model proteins (cytochrome c, carbonic anhydrase, and bovine serum albumin), the tryptic peptides digested by the IMER were analyzed using LC-MS/MS with the dual application of CAD and ETD. As previously shown by others, the dual application of CAD and ETD increased the sequence coverage in comparison with CAD application only. In particular, ETD was very useful for the analysis of highly-protontated peptide cations, e.g., ${\geq}3+$. The combination approach provided the advantages of both trypsin-IMER and CAD/ETD dual tandem mass spectrometry applications, which are rapid digestion (i.e., 10 min), good digestion efficiency, online coupling of trypsin-IMER and liquid chromatography, and high sequence coverage.

Analysis of the Involvement of Chitin-Binding Domain of ChiCW in Antifungal Activity, and Engineering a Novel Chimeric Chitinase with High Enzyme and Antifungal Activities

  • Huang, Chien-Jui;Guo, Shu-Huei;Chung, Shu-Chun;Lin, Yu-Ju;Chen, Chao-Ying
    • Journal of Microbiology and Biotechnology
    • /
    • 제19권10호
    • /
    • pp.1169-1175
    • /
    • 2009
  • An antifungal chitinase, ChiCW, produced by Bacillus cereus 28-9 is effective against conidial germination of Botrytis elliptica, the causal agent of lily leaf blight. ChiCW as a modular enzyme consists of a signal peptide, a catalytic domain, a fibronectin type-III-like domain, and a chitin-binding domain. When two C-terminal domains of ChiCW were truncated, $ChiCW{\Delta}FC$ (lacking the chitin-binding domain and fibronectin type III-like domain) lost its antifungal activity. Since $ChiCW{\Delta}C$ (lacking the chitin-binding domain) could not be expressed in Escherichia coli as $ChiCW{\Delta}FC$ did, a different strategy based on protein engineering technology was designed to investigate the involvement of the chitin-binding domain of ChiCW ($ChBD_{ChiCW}$) in antifungal activity in this study. Because ChiA1 of Bacillus circulans WL-12 is a modular enzyme with a higher hydrolytic activity than ChiCW but not inhibitory to conidial germination of Bo. elliptica and the similar domain composition of ChiA1 and ChiCW, the C-terminal truncated derivatives of ChiA1 were generated and used to construct chimeric chitinases with $ChBD_{ChiCW}$. When the chitin-binding domain of ChiA1 was replaced with $ChBD_{ChiCW}$, the chimeric chitinase named ChiAAAW exhibited both high enzyme activity and antifungal activity. The results indicate that $ChBD_{ChiCW}$ may play an important role in the antifungal activity of ChiCW.

Expression of Nutritionally Well-balanced Protein, AmA1, in Saccharomyces cerevisiae

  • Kim, Tae-Geum;Kim, Ju;Kim, Dae-Hyuk;Yang, Moon-Sik
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제6권3호
    • /
    • pp.173-178
    • /
    • 2001
  • Food yeast, Saccharomyces cerevisiae, is a safe organism with a long history of use for the production of biomass rich in high quality proteins and vitamins. AmA1, a seed storage albumin from Amaranthus hypochondriacus, has a well-balanced amino acid composition and high levels of essential amino acids and offers the possibility of further improving food animal feed additives. In order to find an effective means of expressing AmA1 in yeast, the gene was cloned into an episomal shuttle vector. Four different promoters were tested: the glyceraldehyde-3-phosphate dehydrogenase promoter, galactose dehydrogenase 10 promoter, alcohol dehydrogenase II promoter, and a hybrid ADH2-GPD promoter. The recombinant AmA1 genes were then introduced into the yeast Saccharomyces cerevisiae 2805. Northern and Western blot analyses of the yeast under appropriate conditions revealed that AmA1 was expressed by all four promoters at varying levels. An enzyme-linked immunosorbent assay demonstrated that the amount of AmA1 protein in the recombinant yeast was 1.3-4.3% of the total soluble proteins. The highest expression level was obtained from the hybrid ADH2-GPD promoter.

  • PDF

Design of Smart Polymer Constructs for Use in Microfluidic Diagnostic Assays

  • Hoffman Allan S.
    • 한국고분자학회:학술대회논문집
    • /
    • 한국고분자학회 2006년도 IUPAC International Symposium on Advanced Polymers for Emerging Technologies
    • /
    • pp.42-43
    • /
    • 2006
  • We have prepared random, block and graft copolymers with single or dual sensitivities to various stimuli. We have conjugated these polymers to proteins at random lysine sites or at specific sites designed into the protein by genetic engineering. We are also grafting the smart polymers to the surfaces of nanobeads. We are applying these smart conjugates and smart nanobeads in microfluidic devices for various applications, including diagnostics, affinity separations and enzyme bioprocesses. In this talk I will update our work with these interesting hybrid systems.

  • PDF