• Title/Summary/Keyword: hybrid enzyme

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Isolation and Culture of In Vitro Cultured Populus alba×P. grandidentata Protoplasts (Populus alba×P. grandidentata 조직배양(組織培養) 식물체(植物體) 원형질의 분리(分離)와 배양(培養))

  • Chun, Young Woo
    • Journal of Korean Society of Forest Science
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    • v.71 no.1
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    • pp.45-49
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    • 1985
  • Protoplast-source meterial and enzyme strength had a significant influence on protoplast yield from hybrid poplar, Populus alba ${\times}$ P. grandidentata. The yield of protoplasts from in vitro culture of 1 month-old plantlets was more than that from greenhouse grown 4 month-old stock plant. In vitro cultured plantlets regulary produced more viable protoplasts with E-I enzyme solution (0.5% cellulase and 0.1% macerase) than those with E-II enzyme solution (1.0% cellulase and 0.2% macerase) after overnight incubation. The mean yield of protoplasts from in vitro cultured plantlets was $4{\times}10^6$ with E-I enzyme solution. Cell division was observed in these protoplast cultures after 7-10 days. Protoplast-derived hybrid poplar cells survived over 3 weeks in culture and some continuous cell divisions were evident. Other aspects associated with protoplasts from in vitro cultured plantlet are also discussed.

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Effects of Vitis coignetiae on the Quality and Antihypertension of Vitis hybrid Red Wine (쉐리단 포도주의 항고혈압성과 품질에 머루첨가의 영향)

  • Jang, Jeong-Hoon;Yi, Sung-Hun;Kim, Jae-Ho;Lee, Dae-Hyoung;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.126-132
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    • 2011
  • The goal of this study was to develop a high value Korean red wine possessing antihypertension activity. The effects of some medicinal plants and grapes on the alcohol fermentation process and the angiotensin I-converting enzyme (ACE) inhibitory activity of Vitis hybrid red wine were investigated. Various Vitis hybrid red wines were vinified by the fermentation of a mixture of Vitis hybrid must and some medicinal plants and grapes at $25^{\circ}C$ for 10 days. Of these red wines, the Vitis hybrid-Vitis coignetiae red wine exhibited a high ethanol content of 12.0% and had a good level of acceptability. It also showed a high antihypertensive ACE inhibitory activity of 68.5%. After post-fermentation of 60 days, the ACE inhibitory activities of the Vitis hybrid-Vitis coignetiae red wine exhibited the highest ACE inhibitory activity of 80.7% ($IC_{50}$: 28 mg/mL) and also had the best acceptability. The $C_{18}$ solid phase extracts of the Vitis hybrid-Vitis coignetiae red wine, after 60 days post-fermentation, showed clear antihypertensive effects on spontaneously hypertensive rats. Our results reveal that the Vitis hybrid-Vitis coignetiae red wine has the potential to become a new functional red wine due to its good acceptability and high antihypertensive activity.

Production of Glucoamylase from Hybrid Constructed by Intergenic Nuclear Transfer between Saccharomycopsis sp. and Saccharomyces sp. (핵전이법에 의해 형성된 Saccharomycopsis 속과 Saccharomyces 속의 잡종에서 glucoamylase 생산에 관한 연구)

  • 양영기;임채영;김종권;문명님;이영하
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.182-188
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    • 2001
  • The glucoamylase was purified from the induced culture filtrate of hybrid between Saccharomycopsis sp. and Saccharomycopsis sp. made by nuclear transfer and characterized for some enzyme properties. The enzymewas purified 76-fold in an overall yield of 16% from the culture medium by ammonium sulfate fractionation,Sephadex G-150 gel permeation chromatography and DEAE-Sephadex A-50 ion exchage chromatography.The molecular weight of the purified glucoamylase was estimated to be 57.5 KDa on SDS-polyacrylamidegel electrophoresis and Sephadex G-150 gel permeation chromatography. The purified enzyme was active atpH-5.0 and $40^{\circ}C$. The Km value for soluble starch was 2.6 mg/ml. The enzymatic activity was stimulated inthe presence of TEX>$Ca^{2+}$, EDTA, $Co^{2+}$, $Mg^{2+}$, and $Mn^{2+}$

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The gene encoding guanidinoacetate methyltransferase (GAMT) maps to mouse chromosome 10 near the locus of hesitant mutation affecting male fertility

  • Chae, Young-Jin;Chung, Chan-Ee;Kim, Byung-Jin;Lee, Mun-Han;Lee, Hang
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.50-51
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    • 1998
  • guanidinoacetate methyltransferase (GAMT) catalyzes the last step of creatine biosynthesis in mammals. Creatine plays an important role in cellular energy metabolism in variety of tissues including brain and male reproductive tract. Congenital deficiency of the enzyme leads to a neurologic disorder in humans. We used an interspecific backcross DNA panel to map Gamt to the central region of mouse Chromosome (Chr) 10 near the locus of hesitant mutation affecting male fertility. We assigned the human GAMT gene to Chr 19 by PCR analysis of a human/rodent somatic hybrid cell line DNA panel, and further localized the human gene to Chr 19 at band p13.3 by PCR analysis of a human radiation hybrid DNA panel. Human chr 19p13.3 is homologous to the central part of mouse Chr 10 where mouse Gamt is located. Furthermore, this part of mouse Chr 10 contains mutant loci the phenotype of which is similar to the GAMT deficiency in human.

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Expression analysis of UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT) gene in an interspecific hybrid grape between Vitis ficifolia var. ganebu and Vitis vinifera cv. Muscat of Alexandria

  • Poudel, Puspa Raj;Goto-Yamamoto, Nami;Mochioka, Ryosuke;Kataoka, Ikuo;Beppu, Kenji
    • Plant Biotechnology Reports
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    • v.2 no.4
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    • pp.233-238
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    • 2008
  • Kadainou R-1, an interspecific hybrid grape derived from red (Vitis ficifolia var. ganebu) and white (V. vinifera cv. Muscat of Alexandria) grapes, accumulates high concentrations of anthocyanin in the berry skin. Hence, the expression of uridine 50 -diphosphate (UDP)-glucose:flavonoid 3-O-glucosyltransferase (UFGT), the key enzyme of the anthocyanin pathway, was examined in the berry skin of Kadainou R-1. As information on gene sequences of V. ficifolia var. ganebu and other wild grape species was unavailable, we performed GeneChip hybridization using biotin-labeled genomic deoxyribonucleic acid (DNA) to investigate how the genomic sequences of V. vinifera varieties and that of V. ficifolia var. ganebu differ. The study showed a lower correlation coefficient between V. vinifera cultivars and V. ficifolia var. ganebu than that among V. vinifera cultivars. The sequences of the UFGT gene derived from both parents of the red and white cultivars were sequenced in Kadainou R1 and revealed that both were expressed irrespective of the fact that it was not expressed in the white grape (male parent).

Interaction of GAT1 with Ubiquitin-Specific Protease Usp14 in Synaptic Terminal (GAT1과 ubiquitin-specific protease Usp14의 결합)

  • Seog, Dae-Hyun;Kim, Sang-Jin;Joung, Young-Ju;Yea, Sung-Su;Park, Yeong-Hong;Kim, Moo-Seong;Moon, Il-Soo;Jang, Won-Hee
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1005-1011
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    • 2010
  • $\gamma$-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system. GABA transporters (GATs) control extracellular GABA levels by reuptake of released GABA from the synaptic cleft. However, how GATs are regulated has not yet been elucidated. Here, we used the yeast two-hybrid system to identify the specific binding protein(s) that interacts with the carboxyl (C)-terminal region of GAT1, the major isoform in the brain and find a specific interaction with the ubiquitin-specific protease 14 (Usp14), a deubiquitinating enzyme. Usp14 protein bound to the tail region of GAT1 and GAT2 but not to other GAT members in the yeast two-hybrid assay. The C-terminal region of Usp14 is essential for interaction with GAT1. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to GAT1 specifically co-immunoprecipitated Usp14 from mouse brain extracts. These results suggest that Usp14 may regulate the number of GAT1 at the cell surface.

No Trace of Introduced cpDNA of Pinus thunbergii in Pinus densiflor for. erecta Postulated as an Introgressive Hybrid between Pinus densiflora and Pinus Thunbergii (소나무와 곰솔간 이입교잡종(移入交雜種)으로 추정(推定)되어온 금강송(金剛松)에 있어서 곰솔 cpDNA 의 부재(不在))

  • Hong, Yong-Pyo;Kim, Kyu-Sik;Noh, Eui-Rae;Shin, Eun-Myeong;Kim, Zin-Suh
    • Journal of Korean Society of Forest Science
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    • v.87 no.4
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    • pp.543-548
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    • 1998
  • Portions of chloroplast genes(psbD and rbcL) were amplified from Pinus thunbergii(Japanese black pine : black pine) and Pinus densiflora(Japanese red pine : red pine) by PCR and digested by a restriction enzyme, HaeIII, respectively. Two species specific cpDNA markers were identified. With the observed cpDNA markers, paternal inheritance of cpDNA in pine hybrids was verified in an artificial hybrid family between black pine(Chollanam 37) and red pine(Chungchongbuk 3). On the basis of paternal inheritance of chloroplast genome in a hybrid, 2 portions of cpDNA amplified from 115 individuals of Pinus densiflora for. erecta were screened to detect any traces of black pine specific cpDNA markers in P. densiflora for. erecta which has been postulated as an introgressive hybrid between red pine and black pine(Hyun el al., 1967). All the analyzed individuals of Pinus densiflora for. erects revealed the identical profiles of HaeIII digested psbD and rbcL genes to red pine. This result suggests that there is no introduced chloroplast genome of black pine in Pinus densiflora for. erecta and that there is no concrete evidence of treating P. densiflora for, erecta as an introgressive hybrid between red pine(♀) and black pine(♂).

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Engineered Recombinant PON1-OPH Fusion Hybrids: Potentially Effective Catalytic Bioscavengers against Organophosphorus Nerve Agent Analogs

  • Lee, Nari;Yun, Hyeongseok;Lee, Chan;Lee, Yikjae;Kim, Euna;Kim, Sumi;Jeon, Hyoeun;Yu, Chiho;Rho, Jaerang
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.144-153
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    • 2021
  • Organophosphorus nerve agents (OPNAs), including both G- and V-type nerve agents such as sarin, soman, tabun and VX, are extremely neurotoxic organophosphorus compounds. Catalytic bioscavengers capable of hydrolyzing OPNAs are under development because of the low protective effects and adverse side effects of chemical antidotes to OPNA poisoning. However, these bioscavengers have certain limitations for practical application, including low catalytic activity and narrow specificity. In this study, we generated a fusion-hybrid form of engineered recombinant human paraoxonase 1 (rePON1) and bacterial organophosphorus hydrolase (OPH), referred to as GV-hybrids, using a flexible linker to develop more promising catalytic bioscavengers against a broad range of OPNAs. These GV-hybrids were able to synergistically hydrolyze both G-type OPNA analogs (paraoxon: 1.7 ~ 193.7-fold, p-nitrophenyl diphenyl phosphate (PNPDPP): 2.3 ~ 33.0-fold and diisopropyl fluorophosphates (DFP): 1.4 ~ 22.8-fold) and V-type OPNA analogs (demeton-S-methyl (DSM): 1.9 ~ 34.6-fold and malathion: 1.1 ~ 4.2-fold above) better than their individual enzyme forms. Among the GV-hybrid clones, the GV7 clone showed remarkable improvements in the catalytic activity toward both G-type OPNA analogs (kcat/Km (106 M-1 min-1): 59.8 ± 0.06 (paraoxon), 5.2 ± 0.02 (PNPDPP) and 47.0 ± 6.0 (DFP)) and V-type OPNA analogs (kcat/Km (M-1 min-1): 504.3 ± 48.5 (DSM) and 1324.0 ± 47.5 (malathion)). In conclusion, we developed GV-hybrid forms of rePON1 and bacterial OPH mutants as effective and suitable catalytic bioscavengers to hydrolyze a broad range of OPNA analogs.

The embryological studies on the interspecific hybrid of ginseng plant (Panax ginseng x P. Quiuquefolium) with special references to the seed abortion (인삼의 종간잡종 Panax ginseng x P Quinquefoilium의 발생학적 연구 특히 결실불능의 원인에 관하여)

  • Jong-Kyu Hwang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.5 no.1
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    • pp.69-86
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    • 1969
  • On the growing of the interspecific hybrid ginseng plant, the phenomena of hybrid vigoures are observed in the root, stem, and leaf, but it can not produce seeds favorably since the ovary is abortive in most cases in interspecific hybrid plants. The present investigation was undertaken in an attempt to elucidate the embryological dses of the seed failure in the interspecific hybrid of ginseng (Panax Ginseng ${\times}$ P. Quinque folium). And the results obtained may be summarized as follows. 1). The vegetative growth of the interspecific hybrid ginseng plant is normal or rather vigorous, but the generative growth is extremely obstructed. 2). Even though the generative growth is interrupted the normal development of ovary tissue of flower can be shown until the stage prior to meiosis. 3). The division of the male gameto-genetic cell and the female gameto-genetic cell are exceedingly irregular and some of them are constricted prior to meiosis. 4). At meiosis in the microspore mother cell of the interspecific hybrid, abnormal division is observed in that the univalent chromosome and chromosome bridge occure. And in most cases, metaphasic configuration is principally presented as 23 II+2I, though rarely 22II+4I is also found. 5). Through the process of microspore and pollen formation of F1, the various developmental phases occur even in an anther loclus. 6). Macro, micro and empty pollen grains occur and the functional pollen is very rare. 7). After the megaspore mother cell stage, the rate of ovule development is, on the whole, delayed but the ovary wall enlargement is nearly normal. 8). Degenerating phenomena of ovules occur from the megaspore mother cell stage to 8-nucleate embryo sac stage, and their beginning time of constricting shape is variously different. 9). The megaspore arrangement in the parent is principally of the linear type, though rarely the intermediate type is also observed, whereas various types, viz, linear, intermediate, Tshape, and I shape can be observed in hybrid. 10). After meiosis, three or five megaspore are some times counted. 11). Charazal end megaspore is generally functional in the parents, whereas, in F1, very rarely one of the center megaspores (the second of the third megaspore) grows as an embryo sac mother cell. 12). In accordance with the extent of irregularity or abnormality in meiosis, division of embryo sac nuclei and embryo sac formation cause more nucellus tissue to remain within th, embryo sac. 13). Even if one reached the stage of embryo sac formation, the embryo sac nuclei are always precarious and they can not be disposed to theil proper, respective position. 14). Within the embryo sac, which is lacking the endospermcell, the 4-celled proembryo, linear arrangement, is observed. 15). Through the above respects, the cause of sterile or seed failure of interspecific hybrid would be presumably as follows, By interspecific crossing gene reassortments takes place and the gene system influences the metabolism by the interference of certain enzyme as media. In the F1 plant, the quantity and quality of chemicals produced by the enzyme system and reaction system are entirely different from the case of the parents. Generally, in order to grow, form, and develop naw parts it is necessary to change the materials and energy with reasonable balance, whereas in the F1 plant the metabolic process becomes abnormal or irregular because of the breakdown of the balancing. Thus the changing of the gene-reaction system causes the alteration of the environmental condition of the gameto-genetic cells in the anther and ovule; the produced chemicals cause changes of oxidatio-reduction potential, PH value, protein denaturation and the polarity, etc. Then, the abnormal tissue growing in the ovule and emdryo sac, inhibition of normal development and storage of some chemicals, especially inhibitor, finally lead to sterility or seed failure. Inconclusion, we may presume that the first cause of sterile or seed abortion in interspecific hybrids is the gene reassortment, and the second is the irregularity of the metabolic system, storage of chemicals, especially inhibitor, the growth of abnormal tissue and the change of the polarity etc, and they finally lead to sexual defect, sterility and seed failure.

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Seasonal Expression of OMT Gene in Relation to Lignin biosynthesis in two Poplar Species (종의 포플라수종에서 리그닌생합성에 관계된 OMT유전자의 발현)

  • Park, Young-Goo;Park, Hee Sung;Choi, Jang Won;Sul, Ill Whan;Chung, Il Kyung;Shin, Dong Ill
    • Journal of Life Science
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    • v.8 no.4
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    • pp.443-448
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    • 1998
  • We analyzed lignin content and wxpression of OMT gene during growth season in two hybrid poplar species. OMT gene expression was observed mainy in the developing secondary xylem where major quantity of lignin occurs. Lignin content in the xylem tissue increased as plant resumed growth in the spring and reached the highest in the late August. Change in lignin content was concurrent with that of OMT gene expression, indicating OMT is a key enzyme in lignin biosynthesis.

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