• Title/Summary/Keyword: enzyme transport

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Study of the Function of β-D-fructofuranosidase by Plant Histological Method (식물 조직학적 방법에 의한 β-D-fructofuranosidase의 기능 연구)

  • Donggiun Kim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.4
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    • pp.587-592
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    • 2023
  • Plant growth is regulated by a variety of factors, including organic matter availability. Organic nutrients are carbohydrate molecules from photosynthetic products produced by tissues associated with carbon and energy fixation called "sources". These compounds flow through plant vascular bundles into non-photosynthetic or growing tissues called "sinks". Among these possible compounds, the disaccharide fructosyl glucose, sucrose, is the most representative. During the transport of sucrose, the pathway from the source to the sinks can include hydrolysis of sucrose into glucose and fructose derivatives or direct transfer of sucrose. Among the enzymes involved in this, β-D-fructofuranosidase is the most important. Soluble neutral β-D-fructofuranosidase, one of several isoenzymes, is located in intracellular protoplasts and helps plant cells metabolize sucrose to produce energy. In order to track the activity of this enzyme during the course of plant growth, histological methods were used for the most effective immunolocalization. As a result, the activity was higher in the phloem and epidermis than in the mesophyll tissue in the leaf. In the growing stem, activity was high in the phloem, epidermis, and cortex. The activity of the root, which is a sink tissue, was high in all parts, but especially the highest in the root tip part. It is thought that this is because it helps unloading of sucrose in sink tissues that require sucrose degradation and plays a role in hydrolysising sucrose.

Field Tests for Assessing the Bioremediation Feasibility of a Trichloroethylene-Contaminated Aquifer (관측정 자연표류 실험을 통한 트리클로로에틸렌(Trichloroethylene) 오염 지하수의 생물학적 복원 타당성 연구)

  • Kim Young;Kim Jin-Wook;Ha Chul-Yoon;Kim Nam-Hee;Hong Kwang-Pyo;Kwon Soo-Yul;Ahn Young-Ho;Ha Joon-Su;Park Hoo-Won
    • Journal of Soil and Groundwater Environment
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    • v.10 no.3
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    • pp.38-45
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    • 2005
  • The feasibility of stimulating in situ aerobic cometabolic activity of indigenous microorganisms was investigated in a trichloroethylene (TCE)-contaminated aquifer. A series of single-well natural drift tests (SWNDTs) was conducted by injecting site groundwater amended with a bromide tracer and combinations of toluene, oxygen, nitrate, ethylene and TCE into an existing monitoring well and by sampling the same well over time. Three field tests, Push-pull Transport Test, Drift Biostimulation Test, and Drift Surrogate Activity Test, were performed in sequence. Initial rate of toluene degradation was much faster than the rate of bromide dilution resulting from natural groundwater drift, indicating stimulation of indigenous toluene-oxidizing microorganisms. Transformation of ethylene, a surrogate probing overall activity of TCE transformation, was also observed, and its transformation results in the production of ethylene oxide, suggesting that some tolueneoxidizing microorganisms stimulated may express a orthomonooxygenase enzyme. Also in situ transformation of TCE was confirmed by greater retardation of TCE than bromide after the stimulation of toluene-oxidizing microorganisms. These results indicate that, in this environment, toluene and oxygen additions stimulated the growth and aerobic cometabolic activity of indigenous microorganisms expressing orthomonooxygenase enzymes. The simple, low-cost field test method presented in this study provides an effective method for conducting rapid field assessments and pilot testing of aerobic cometabolism, which has previously hindered application of this technology to groundwater remediation.

Induction of Phase I, II and III Drug Metabolism/Transport by Xenobiotics

  • Xu Chang Jiang;Li Christina YongTao;Kong AhNg Tony
    • Archives of Pharmacal Research
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    • v.28 no.3
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    • pp.249-268
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    • 2005
  • Drug metabolizing enzymes (DMEs) play central roles in the metabolism, elimination and detoxification of xenobiotics and drugs introduced into the human body. Most of the tissues and organs in our body are well equipped with diverse and various DMEs including phase I, phase II metabolizing enzymes and phase III transporters, which are present in abundance either at the basal unstimulated level, and/or are inducible at elevated level after exposure to xenobiotics. Recently, many important advances have been made in the mechanisms that regulate the expression of these drug metabolism genes. Various nuclear receptors including the aryl hydrocarbon receptor (AhR), orphan nuclear receptors, and nuclear factor-erythoroid 2 p45-related factor 2 (Nrf2) have been shown to be the key mediators of drug-induced changes in phase I, phase II metabolizing enzymes as well as phase III transporters involved in efflux mechanisms. For instance, the expression of CYP1 genes can be induced by AhR, which dimerizes with the AhR nuclear translocator (Arnt) , in response to many polycyclic aromatic hydrocarbon (PAHs). Similarly, the steroid family of orphan nuclear receptors, the constitutive androstane receptor (CAR) and pregnane X receptor (PXR), both heterodimerize with the ret-inoid X receptor (RXR), are shown to transcriptionally activate the promoters of CYP2B and CYP3A gene expression by xenobiotics such as phenobarbital-like compounds (CAR) and dexamethasone and rifampin-type of agents (PXR). The peroxisome proliferator activated receptor (PPAR), which is one of the first characterized members of the nuclear hormone receptor, also dimerizes with RXR and has been shown to be activated by lipid lowering agent fib rate-type of compounds leading to transcriptional activation of the promoters on CYP4A gene. CYP7A was recognized as the first target gene of the liver X receptor (LXR), in which the elimination of cholesterol depends on CYP7A. Farnesoid X receptor (FXR) was identified as a bile acid receptor, and its activation results in the inhibition of hepatic acid biosynthesis and increased transport of bile acids from intestinal lumen to the liver, and CYP7A is one of its target genes. The transcriptional activation by these receptors upon binding to the promoters located at the 5-flanking region of these GYP genes generally leads to the induction of their mRNA gene expression. The physiological and the pharmacological implications of common partner of RXR for CAR, PXR, PPAR, LXR and FXR receptors largely remain unknown and are under intense investigations. For the phase II DMEs, phase II gene inducers such as the phenolic compounds butylated hydroxyanisol (BHA), tert-butylhydroquinone (tBHQ), green tea polyphenol (GTP), (-)-epigallocatechin-3-gallate (EGCG) and the isothiocyanates (PEITC, sul­foraphane) generally appear to be electrophiles. They generally possess electrophilic-medi­ated stress response, resulting in the activation of bZIP transcription factors Nrf2 which dimerizes with Mafs and binds to the antioxidant/electrophile response element (ARE/EpRE) promoter, which is located in many phase II DMEs as well as many cellular defensive enzymes such as heme oxygenase-1 (HO-1), with the subsequent induction of the expression of these genes. Phase III transporters, for example, P-glycoprotein (P-gp), multidrug resistance-associated proteins (MRPs), and organic anion transporting polypeptide 2 (OATP2) are expressed in many tissues such as the liver, intestine, kidney, and brain, and play crucial roles in drug absorption, distribution, and excretion. The orphan nuclear receptors PXR and GAR have been shown to be involved in the regulation of these transporters. Along with phase I and phase II enzyme induction, pretreatment with several kinds of inducers has been shown to alter the expression of phase III transporters, and alter the excretion of xenobiotics, which implies that phase III transporters may also be similarly regulated in a coordinated fashion, and provides an important mean to protect the body from xenobiotics insults. It appears that in general, exposure to phase I, phase II and phase III gene inducers may trigger cellular 'stress' response leading to the increase in their gene expression, which ultimately enhance the elimination and clearance of these xenobiotics and/or other 'cellular stresses' including harmful reactive intermediates such as reactive oxygen species (ROS), so that the body will remove the 'stress' expeditiously. Consequently, this homeostatic response of the body plays a central role in the protection of the body against 'environmental' insults such as those elicited by exposure to xenobiotics.

A study of the lipoprotein lipase inhibitory mechanism of Poncirus trifoliata water extracts (탱자 (Poncirus trifoliata)의 lipoprotein lipase 억제메커니즘)

  • Lee, Sung Mee;Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Tae Woo;Choe, Myeon
    • Journal of Nutrition and Health
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    • v.48 no.1
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    • pp.9-18
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    • 2015
  • Purpose: Poncirus trifoliata has been reported to have anti-inflammatory, antioxidant, and immune activities. However, its anti-obesity activity and the mechanism by which the water extract of dried, immature fruit of Poncirus trifoliata (PF-W) acts are not clear. This study suggests a potential mechanism associated with the anti-obesity activity of PF-W. Methods: We measured the effect of PF-W on lipoprotein lipase (LPL) regulation using enzyme-linked immunosorbent assay (ELISA) and an activity assay. The LPL regulation mechanism was examined by reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression of biomarkers related to protein transport and by western blot for analysis of the protein expression of the transcription factor CCAAT-enhancer-binding protein ($C/EBP{\beta}$). Results: The total polyphenol and flavonoid content of PF-W was $52.15{\pm}4.02$ and $6.56{\pm}0.47mg/g$, respectively. PF-W treatment decreased LPL content in media to $58{\pm}5%$ of that in control adipocyte media, and increased LPL content to $117{\pm}3.5%$ of that in control adipocytes, but did not affect the mRNA expression of LPL. PF-W also increased the mRNA expression of sortilin-related receptor (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL, in a concentration- and time-dependent manner. Finally, cell fractionation revealed that PF-W treatment induced the expression of $C/EBP{\beta}$, a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. Conclusion: The LPL secretion and activity assay showed PF-W to be an LPL secretion inhibitor, and these results suggest the potential mechanism of PF-W involving inhibition of LPL secretion through $C/EBP{\beta}$-mediated induction of SorLA expression.

Effects of SNP Markers of the Apolipoprotein E (APOE) Gene on Meat Quantity and Quality Traits in Korean Cattle (한우 아포지단백질 E (APOE) 유전자의 SNP Marker가 육량 및 육질형질에 미치는 영향)

  • Shin, Ki-Hyun;Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.108-113
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    • 2009
  • Apolipoprotein E (APOE) is a plasma lipoprotein in mammals and plays an important role in the transport and metabolism of lipids such as phospholipids and triglycerides. Therefore, the APOE gene could be a candidate gene controlling lipid metabolism in beef cattle. This study was performed to identify single nucleotide polymorphisms (SNP) in the APOE gene and to investigate the effects of SNP genotype on the carcass traits such as meat quantity and quality in Korean cattle. For PCR amplification, pooled DNA made from unrelated 60 individuals was prepared and primer pairs were designed based on the cDNA sequence of exon 4 region of the bovine APOE gene. A SNP was identified at position 2034 (T/C substitution) of the exon 4 region in the APOE gene. PCR-RFLP procedure with restriction enzyme ACC I was developed for determining the SNP genotype for each of a total of 309 animals with pedigree information and performance records through the national progeny testing program. The frequencies of the genotypes TT, TC and CC were 10.9, 46.9 and 42.2%. Gene frequencies were 0.344 for T allele and 0.656 for C allele. The g.2034T>C SNP genotype showed a significant effect (p<0.05) on dressing percentage and meat color, respectively. Animals with the TT genotype showed higher dressing percentage than those with the CC genotype, and TT genotype had desirable meat color compared with CC genotype. These results suggest that the g.2034T>C SNP genotype of the APOE gene may be useful as a DNA marker for meat quantity index and dressing percentage in Korean cattle.

Objectives and Contents of Basic Medical Sciences in Nursing Education (간호학 교육에서 기초의과학 교과목별 목표와 내용에 대한 연구)

  • 최명애;신기수
    • Journal of Korean Academy of Nursing
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    • v.29 no.6
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    • pp.1455-1468
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    • 1999
  • The purpose of this study was to investigate the objectives and contents of basic medical sciences at department of nursing in college of nursing, and junior college of nursing, thus ultimately providing the basic data to standardize the curriculum of the basic medical sciences in nursing education. Seventy eight professors who were in charge of teaching basic medical sciences to at 22 colleges of nursing/ department of nursing, and 20 junior colleges of nursing responded to the questionnaires that consisted of the questions regarding objectives and contents, of basic medical sciences. Based on the description of objectives, the description related to nursing, nurse, nursing science was cathegorized as on objective applicable to nursing science, the description related to medicine or clinical medicine as medical model, the description without description related to medicine was cathegorized as knowledge acquisition. The number of schools corresponding to each category were summerized in descending order. The objectives of basic medical sciences were categorized by concepts and number of schools corresponding to the categorized concept. The findings of the study are as follows ; 1. The subjects of basic medical science identified were physiology, anatomy, biochemistry, pathology, microbiology, and pharmacology in most colleges of nursing and junior colleges. Two colleges of nursing/department of nursing (9.1%) and 19 junior colleges of nursing(95%) did not offer biochemistry, 1 college of nursing /department of nursing(5%) did not offer pathology & pharmacology. 2 junior colleges of nursing (10%) did not offer pharmacology, 1 junior college of nursing(5%) did not offer pathology. The other 1 junior college of nursing did not offer microbiology. 2. Objectives of physiology were to acquire knowledge and understanding on human function in both 6 (50%) colleges and 5 junior colleges. Objectives of anatomy were to acquire knowledge on human structure in both 4 (57%) colleges and 2 (50%) junior colleges; knowledge applicable to nursing sciences in both 3 (42.8%) colleges and 2 (50%) junior colleges. Objectives of biochemistry was to obtain knowledge and understanding on biochemistry, and understanding of basic concepts about biochemistry. Objectives of pathology were to obtain knowledge and understanding on pathology in both 4 (57.1%) colleges and 5(62.5%) junior colleges. Objectives of microbiology were to acquire knowledge and understanding on microbiology in both 5(83.8%) colleges and 6(85.7%) junior colleges. Objectives of pharmacology were to acquire knowledge on pharmacology in both 7(100%) colleges and 8(100%) junior colleges. 3. Contents of physiology in 19 (100%) schools were membrane transport, digestion, circulation, nervous system and respiration. In 16(84.2%) were kidney and muscle, that in 13(68.4%) were endocrine physiology. In 11(57.9%) were introduction and that in 9(47.4%) were structure and function of cells. Contents of anatomy in 11(100%) schools were skeletal system, muscle system, digestive system, circulatory system, concepts regarding human structure. In 10(90.9%) schools were endocrine system and nervous system, and in 5(45.5%) schools were blood, urinary system and cell. Contents of biochemistry in 6(100%) schools were history of biochemistry, body regulating factor, bioenergy, health and nutrition, nutrition of cell, energy production system. In 5(83.3%) schools were metabolism of protein and carbohydrate and enzyme, and in 3(50%) schools were metabolism of energy and fat. Contents of microbiology in 13(100%) schools were environment and influenc of bacteria, virus, G(-) rods, purulent cocci, G(+) rods. In 10 (76.9%) were immunity, diphtheria, enterobacteria, and in 9(69.2%) were spirochete, rickettsia and clamydia, and that in 6(46.2%) were sterilization and disinfection. Contents of pathology in 14(100%) schools were cell injury and adaptation, inflammation, respiratory diseases, circulatory diseases. In 10(71.4%) were neurological disorders, in 8(57.1%) were immunity and disease, and in 7 (50%) were tumor and progressive changes. Contents of pharmacology in 15(100%) were cardivascular drugs, introduction to pharmacology, hypnotics, analgesics, local anesthetics, an ticonvulsants. In 12(80%) were drugs activity on sympathetic and parasympathetic nervous system, and in 11(73%) were sulfa drugs, antibiotics, drug abuse and addiction.

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Role of Citrate Synthase in Acetate Utilization and Protection from Stress-Induced Apoptosis

  • Lee, Yong-Joo;Kang, Hong-Yong;Maeng, Pil Jae
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.39-41
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    • 2008
  • The yeast Saccharomyces cerevisiae has been shown to contain three isoforms of citrate synthase (CS). The mitochondrial CS, Cit1, catalyzes the first reaction of the TCA cycle, i.e., condensation of acetyl-CoA and oxaloacetate to form citrate [1]. The peroxisomal CS, Cit2, participates in the glyoxylate cycle [2]. The third CS is a minor mitochondrial isofunctional enzyme, Cit3, and related to glycerol metabolism. However, the level of its intracellular activity is low and insufficient for metabolic needs of cells [3]. It has been reported that ${\Delta}cit1$ strain is not able to grow with acetate as a sole carbon source on either rich or minimal medium and that it shows a lag in attaining parental growth rates on nonfermentable carbon sources [2, 4, 5]. Cells of ${\Delta}cit2$, on the other hand, have similar growth phenotype as wild-type on various carbon sources. Thus, the biochemical basis of carbon metabolism in the yeast cells with deletion of CIT1 or CIT2 gene has not been clearly addressed yet. In the present study, we focused our efforts on understanding the function of Cit2 in utilizing $C_2$ carbon sources and then found that ${\Delta}cit1$ cells can grow on minimal medium containing $C_2$ carbon sources, such as acetate. We also analyzed that the characteristics of mutant strains defective in each of the genes encoding the enzymes involved in TCA and glyoxylate cycles and membrane carriers for metabolite transport. Our results suggest that citrate produced by peroxisomal CS can be utilized via glyoxylate cycle, and moreover that the glyoxylate cycle by itself functions as a fully competent metabolic pathway for acetate utilization in S. cerevisiae. We also studied the relationship between Cit1 and apoptosis in S. cerevisiae [6]. In multicellular organisms, apoptosis is a highly regulated process of cell death that allows a cell to self-degrade in order for the body to eliminate potentially threatening or undesired cells, and thus is a crucial event for common defense mechanisms and in development [7]. The process of cellular suicide is also present in unicellular organisms such as yeast Saccharomyces cerevisiae [8]. When unicellular organisms are exposed to harsh conditions, apoptosis may serve as a defense mechanism for the preservation of cell populations through the sacrifice of some members of a population to promote the survival of others [9]. Apoptosis in S. cerevisiae shows some typical features of mammalian apoptosis such as flipping of phosphatidylserine, membrane blebbing, chromatin condensation and margination, and DNA cleavage [10]. Yeast cells with ${\Delta}cit1$ deletion showed a temperature-sensitive growth phenotype, and displayed a rapid loss in viability associated with typical apoptotic hallmarks, i.e., ROS accumulation, nuclear fragmentation, DNA breakage, and phosphatidylserine translocation, when exposed to heat stress. Upon long-term cultivation, ${\Delta}cit1$ cells showed increased potentials for both aging-induced apoptosis and adaptive regrowth. Activation of the metacaspase Yca1 was detected during heat- or aging-induced apoptosis in ${\Delta}cit1$ cells, and accordingly, deletion of YCA1 suppressed the apoptotic phenotype caused by ${\Delta}cit1$ mutation. Cells with ${\Delta}cit1$ deletion showed higher tendency toward glutathione (GSH) depletion and subsequent ROS accumulation than the wild-type, which was rescued by exogenous GSH, glutamate, or glutathione disulfide (GSSG). Beside Cit1, other enzymes of TCA cycle and glutamate dehydrogenases (GDHs) were found to be involved in stress-induced apoptosis. Deletion of the genes encoding the TCA cycle enzymes and one of the three GDHs, Gdh3, caused increased sensitivity to heat stress. These results lead us to conclude that GSH deficiency in ${\Delta}cit1$ cells is caused by an insufficient supply of glutamate necessary for biosynthesis of GSH rather than the depletion of reducing power required for reduction of GSSG to GSH.

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Current Status of the Research on the Postharvest Technology of Melon(Cucumis melo L.) (멜론(Cucumis melo L.) 수확 후 관리기술 최근 연구 동향)

  • Oh, Su-Hwan;Bae, Ro-Na;Lee, Seung-Koo
    • Food Science and Preservation
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    • v.18 no.4
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    • pp.442-458
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    • 2011
  • Among Cucubitaceae, melon (Cucumis melo) is one of the most diversified fruits, with various forms, sizes, pulps, and peel colors, In addition, it is a commercially important crop because of its high sweetness, deep flavor, and abundant juice. In the species, there are both climacteric and non-climacteric melons depending on the respiration and ethylene production patterns after harvest. Ethylene is also considered a crucial hormone for determining sex expression, Phytohormones other than ethylene interact and regulate ripening, There are some indices that can be used to evaluate the optimum harvest maturity. The harvest time can be estimated after the pollination time, which is the most commonly used method of determining the harvest maturity of the fruit. Besides the physiological aspects, the biochemical alterations, including those of sweetness, firmness, flavor, color, and rind, contribute to the overall fruit quality. These changes can be categorized based on the ethylene-dependent and ethylene-independent phenomena due to the ethylene-suppressed transgenic melon. After harvest, the fruits are precooled to $10^{\circ}C$ to reduce the field heat, after which they are sized and packed. The fruits can be treated with hot water ($60^{\circ}C$ for 60 min) to prevent the softening of the enzyme activity and microorganisms, and with calcium to maintain their firmness. 1-methylenecyclopropene (1-MCP) treatment also maintains their storability by inhibiting respiration and ethylene production. The shelf life of melon is very short even under cold storage, like other cucurbits, and it is prone to obtaining chilling injury under $10^{\circ}C$. In South Korea, low-temperature ($10^{\circ}C$) storage is known to be the best storage condition for the fruit. For long-time transport, CA storage is a good method of maintaining the quality of the fruit by reducing the respiration and ethylene. For fresh-cut processing, washing with a sanitizing agent and packing with plastic-film processing are needed, and low-temperature storage is necessary. The consumer need and demand for fresh-cut melon are growing, but preserving the quality of fresh-cut melon is more challenging than preserving the quality of the whole fruit.

The Heterotrimeric Kinesin-2 Family Member KIF3A Directly Binds to Creatine Kinase B (Heterotrimeric kinesin-2의 KIF3A와 creatine kinase B의 결합)

  • Jeong, Young Joo;Park, Sung Woo;Seo, Mi Kyoung;Kim, Sang-Jin;Lee, Won Hee;Kim, Mooseong;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.31 no.3
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    • pp.257-265
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    • 2021
  • Heterotrimeric kinesin-2 is a molecular motor protein of the kinesin superfamily (KIF) that moves along a microtubule plus-end directed motor protein. It consists of three different motor subunits (KIF3A, KIF3B, and KIF3C) and a kinesin-associated protein 3 (KAP3) that form a heterotrimeric complex. Heterotrimeric kinesin-2 interacts with many different binding proteins through the cargo-binding domain of the KIF3s. The activity of heterotrimeric kinesin-2 is regulated to ensure that the cargo is directed to the right place at the right time. How this regulation occurs, however, remains in question. To identify the regulatory proteins for heterotrimeric kinesin-2, we performed yeast two-hybrid screening and found a specific interaction with creatine kinase B (CKB), which is the brain isoform of cytosolic creatine kinase enzyme. CKB bound to the cargo-binding domain of KIF3A but did not interact with the KIF3B, KIF5B, or KAP3 in the yeast two-hybrid assay. The carboxyl (C)-terminal region of CKB is essential for the interaction with KIF3A. Another protein kinase, CaMKIIa, interacted with KIF3A, but GSK3a did not interact with KIF3A in the yeast two-hybrid assay. KIF3A interacted with GST-CKB-C but not with GSK-CKB-N or GST alone. When co-expressed in HEK-293T cells, CKB co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. These results suggest that the CKB-KIF3A interaction may regulate the cargo transport of heterotrimeric kinesin-2 under energy-compromised conditions in cells.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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