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Sugar-responsive transcription factor bZIP3 affects leaf shape in Arabidopsis plants. 糖反应转录因子bZIP3影响拟南芥叶片形状。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0410A
Miho Sanagi, Yu Lu, Shoki Aoyama, Yoshie Morita, Nobutaka Mitsuda, Miho Ikeda, M. Ohme-Takagi, Takeo Sato, J. Yamaguchi
Sugars are essential for plant metabolism, growth and development. Plants must therefore manage their growth and developmental processes in response to sugar availability. Sugar signaling pathways constitute a complicated molecular network and are associated with global transcriptional regulation. However, the molecular mechanisms underlying sugar signaling remain largely unclear. This study reports that the protein basic-region leucine zipper 3 (bZIP3) is a novel sugar-responsive transcription factor in Arabidopsis plants. The expression of bZIP3 was rapidly repressed by sugar. Genetic analysis indicated that bZIP3 expression was modulated by the SNF1-RELATED KINASE 1 (SnRK1) pathway. Moreover, transgenic plants overexpressing bZIP3 and dominant repressor form bZIP3-SRDX showed aberrant shaped cotyledons with hyponastic bending. These findings suggest that bZIP3 plays a role in plant responses to sugars and is also associated with leaf development.
糖是植物代谢、生长和发育所必需的。因此,植物必须管理其生长和发育过程,以响应糖的可用性。糖信号通路构成了一个复杂的分子网络,并与全球转录调控有关。然而,糖信号传导的分子机制仍不清楚。本研究报道了蛋白基区亮氨酸拉链3 (bZIP3)是拟南芥植物中一种新的糖应答转录因子。bZIP3的表达被糖迅速抑制。遗传分析表明,bZIP3的表达受SNF1-RELATED KINASE 1 (SnRK1)通路的调控。此外,过表达bZIP3和优势抑制因子bZIP3- srdx的转基因植株子叶形状异常,并出现下凸弯曲。这些发现表明,bZIP3在植物对糖的反应中起作用,并与叶片发育有关。
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引用次数: 9
ARF GTPase machinery at the plasma membrane regulates auxin transport-mediated plant growth. 质膜上的ARF GTP酶机制调节生长素转运介导的植物生长。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0312A
S. Naramoto, J. Kyozuka
VAN3 is a plant ACAP-type ADP-ribosylation factor-GTPase activating protein (ARF-GAP) that regulates auxin transport-mediated plant morphogenesis such as continuous venation and lateral root development in Arabidopsis. Previous studies suggested that VAN3 localizes at the plasma membrane (PM) and intracellular structures. However, the role of PM localization in mediating the van3 mutant phenotype is not clear. Here we performed subcellular localization analysis of VAN3 and its regulators CVP2 and VAB to determine their endogenous functions. We found that GFP-tagged CVP2 and VAB preferentially localize at the PM in stably transformed plants. We determined that transgenic plants with lower expression levels of GFP- or mRFP-tagged VAN3 displayed PM localization, which was sufficient to rescue the van3 mutant. Functional VAN3-mRFP and VAB-GFP colocalized at PMs. The van3 mutant phenotype was suppressed by mutation of VAN7/GNOM, which encodes an ARF-GEF that localizes at the PM and Golgi apparatus. These combined results suggest that ARF-GTPase machinery at the PM regulates auxin transport-mediated plant growth and development.
VAN3是一种植物acap型adp -核糖基化因子- gtpase激活蛋白(ARF-GAP),在拟南芥中调节生长素运输介导的植物形态发生,如连续脉化和侧根发育。先前的研究表明,VAN3定位于质膜(PM)和细胞内结构。然而,PM定位在介导van3突变表型中的作用尚不清楚。本研究对VAN3及其调控因子CVP2和VAB进行了亚细胞定位分析,以确定其内源性功能。我们发现gfp标记的CVP2和VAB在稳定转化的植物中优先定位于PM。我们确定GFP-或mrfp -标记的VAN3表达水平较低的转基因植物显示PM定位,这足以拯救VAN3突变体。功能性VAN3-mRFP和VAB-GFP共定位于pm。van3突变表型被VAN7/GNOM突变抑制,该突变编码位于PM和高尔基体的ARF-GEF。这些综合结果表明,PM上的ARF-GTPase机制调节生长素运输介导的植物生长和发育。
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引用次数: 11
Characterization of rice KT/HAK/KUP potassium transporters and K+ uptake by HAK1 from Oryza sativa. 水稻KT/HAK/KUP钾转运蛋白的特性及水稻HAK1对K+的吸收。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0308A
Tomoyuki Okada, Sousuke Yamane, M. Yamaguchi, K. Kato, A. Shinmyō, Yuta Tsunemitsu, K. Iwasaki, D. Ueno, T. Demura
Plant high-affinity K+ (HAK) transporters are divided into four major clusters. Cluster I transporters, in particular, are thought to have high-affinity for K+. Of the 27 HAK genes in rice, eight HAK transporters belong to cluster I. In this study, we investigated the temporal expression patterns during K+ deficiency and K+ transport activity of these eight HAK transporters. The expression of seven HAK genes except OsHAK20 was detected. Expression of OsHAK1, OsHAK5 and OsHAK21 was induced in response to K+ deficiency; however, that of other genes was not. Six of the eight HAK transporters-OsHAK1, OsHAK5, OsHAK19, OsHAK20, OsHAK21, and OsHAK27-complemented the K+-transporter-deficient yeast or bacterial strain. Further, the yeast cells expressing OsHAK1 were more sensitive to Na+ than those expressing OsHAK5. Mutant analysis showed that the high-affinity K+ uptake activity was almost undetectable in oshak1 mutants in a low-K+ medium (0.02 mM). In addition, the high-affinity K+ uptake activity of wild-type plants was inhibited by mild salt stress (20 mM NaCl); however, Na+ permeability of OsHAK1 was not detected in Escherichia coli cells. The high-affinity K+ uptake activity by leaf blades was detected in wild-type plants, while it was not detected in oshak1 mutants. Our results suggest that OsHAK1 and OsHAK5 are the two important components of cluster I corresponding to low-K+ conditions, and that the transport activity of OsHAK1, unlike that of OsHAK5, is sensitive to Na+. Further, OsHAK1 is suggested to involve in foliar K+ uptake.
植物高亲和力K+转运蛋白可分为四大类。特别是簇I转运蛋白被认为对K+具有高亲和力。在水稻的27个HAK基因中,有8个HAK转运蛋白属于I簇。在本研究中,我们研究了K+缺乏期间的时间表达模式和这8个HAK转运蛋白的K+转运活性。检测到除OsHAK20外的7个HAK基因的表达。OsHAK1、OsHAK5和OsHAK21的表达是响应K+缺乏而诱导的;然而,其他基因则不然。八种HAK转运蛋白OsHAK1、OsHAK5、OsHAK19、OsHAK20、OsHAK21和OsHAK27中的六种补充了缺乏K+转运蛋白的酵母或细菌菌株。此外,表达OsHAK1的酵母细胞比表达OsHAK5的酵母细胞对Na+更敏感。突变体分析表明,在低K+培养基(0.02 mM)。此外,野生型植物的高亲和力K+吸收活性受到轻度盐胁迫的抑制(20 mM NaCl);但是在大肠杆菌细胞中未检测到OsHAK1的Na+渗透性。叶片对K+的高亲和力吸收活性在野生型植物中检测到,而在oshak1突变体中没有检测到。我们的结果表明,OsHAK1和OsHAK5是簇I的两个重要组成部分,对应于低K+条件,并且OsHAK1的转运活性不同于OsHAK5,对Na+敏感。此外,OsHAK1被认为参与了叶片K+的吸收。
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引用次数: 20
The functional balance between the WUSCHEL-RELATED HOMEOBOX1 gene and the phytohormone auxin is a key factor for cell proliferation in Arabidopsis seedlings. WUSCHEL相关同源物X1基因和植物激素生长素之间的功能平衡是拟南芥幼苗细胞增殖的关键因素。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0427A
Miyuki T Nakata, Toshiaki Tameshige, M. Takahara, Nobutaka Mitsuda, K. Okada
The WUSCHEL-RELATED HOMEOBOX1 (WOX1) transcription factor and its homolog PRESSED FLOWER (PRS) are multifunctional regulators of leaf development that act as transcriptional repressors. These genes promote cell proliferation under certain conditions, but the related molecular mechanisms are not well understood. Here, we present a new function for WOX1 in cell proliferation. To identify the WOX1 downstream genes, we performed a microarray analysis of shoot apices of transgenic Arabidopsis thaliana lines harboring [35Sp::WOX1-glucocorticoid receptor (GR)] in which the WOX1 function was temporarily enhanced by dexamethasone. The downregulated genes were significantly enriched for the Gene Ontology term "response to auxin stimulus", whereas the significantly upregulated genes contained auxin transport-associated PIN1 and AUX1 and the auxin response factor MP, which are involved in formation of auxin response maxima. Simultaneous treatments of synthetic auxin and dexamethasone induced the formation of green compact calli and the unorganized proliferation of cells in the hypocotyl. A microarray analysis of 35Sp::WOX1-GR plants treated with indole-3-acetic acid and dexamethasone revealed that WOX1 and auxin additively influenced their common downstream genes. Furthermore, in the presence of an auxin-transport inhibitor, cell proliferation during leaf initiation was suppressed in the prs mutant but induced in a broad region of the peripheral zone of the shoot apical meristem in the ectopic WOX1-expressing line FILp::WOX1. Thus, our results clarify the additive effect of WOX1/PRS and auxin on their common downstream genes and highlight the importance of the balance between their functions in controlling cell proliferation.
wuschell相关的HOMEOBOX1 (WOX1)转录因子及其同源物PRESSED FLOWER (PRS)是叶片发育的多功能调控因子,可作为转录抑制因子。这些基因在一定条件下促进细胞增殖,但相关的分子机制尚不清楚。在这里,我们提出了WOX1在细胞增殖中的新功能。为了鉴定WOX1下游基因,我们对含有[35Sp::WOX1-糖皮质激素受体(GR)]的转基因拟南芥植株的茎尖进行了微阵列分析,其中WOX1的功能被地塞米松暂时增强。在基因本体术语“生长素刺激反应”中,下调基因显著富集,而生长素转运相关的PIN1、AUX1和生长素反应因子MP则显著上调,参与生长素反应最大值的形成。合成生长素和地塞米松同时处理可诱导绿色致密愈伤组织的形成和下胚轴细胞的无组织增殖。对经吲哚-3-乙酸和地塞米松处理的35Sp::WOX1- gr植株进行微阵列分析发现,WOX1和生长素共同影响了它们的下游基因。此外,在生长素运输抑制剂的存在下,prs突变体在叶片形成过程中的细胞增殖受到抑制,但在异位WOX1表达系FILp::WOX1的茎尖分生组织的外围区却受到了广泛的诱导。因此,我们的研究结果阐明了WOX1/PRS和生长素对其共同下游基因的加性作用,并强调了它们之间的功能平衡在控制细胞增殖中的重要性。
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引用次数: 15
Control of plant growth and development by overexpressing MAP3K17, an ABA-inducible MAP3K, in Arabidopsis. 通过在拟南芥中过表达MAP3K17(一种ABA诱导的MAP3K)来控制植物生长和发育。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0412A
D. Matsuoka, Kaori Soga, Takuto Yasufuku, T. Nanmori
Abscisic acid (ABA) plays an important role in plant growth, development, and stress responses. ABA regulates many aspects of plant growth and development, including seed maturation, dormancy, germination, the transition from vegetative to reproductive growth, leaf senescence and responses to environmental stresses, such as drought, high salinity and cold. It is also known that mitogen-activated protein kinase (MAPK) cascades function in ABA signaling. Recently, we and another group have identified the ABA-inducible MAP3Ks MAP3K17 and MAP3K18 as the upstream MAP3Ks of MKK3, implicating the MAP3K17/18-MKK3-MPK1/2/7/14 cascade in ABA signaling. It has also been reported that overexpression of MAP3K18 in Arabidopsis causes an early leaf senescence phenotype, ABA hypersensitive stomata closing, and drought tolerance. In this study, we generated transgenic plants overexpressing MAP3K17 (35S:MAP3K17) and its kinase-inactive form (35S:MAP3K17KN). The bolting of 35S:MAP3K17 was earlier than WT, and the fresh weights of the seedlings were smaller, whereas 35S:MAP3K17KN showed the opposite phenotype. These results indicate that the transition from vegetative to reproductive growth can be regulated by overexpression of MAP3K17 and its kinase-inactive form. Moreover, 35S:MAP3K17 showed lower sensitivity to ABA during post-germinated growth, whereas 35S:MAP3K17 KN showed the opposite phenotype, suggesting the negative roles of MAP3K17 in the response to ABA. Our work provides the possibility to regulate plant growth and development by the genetic manipulation of ABA-induced MAPK cascades, leading to improved crop growth and productivity.
脱落酸(ABA)在植物生长发育和胁迫反应中起着重要作用。ABA调节植物生长发育的许多方面,包括种子成熟、休眠、发芽、从营养生长到生殖生长的过渡、叶片衰老以及对干旱、高盐度和寒冷等环境胁迫的反应。众所周知,丝裂原活化蛋白激酶(MAPK)级联在ABA信号传导中发挥作用。最近,我们和另一个小组已经确定ABA诱导型MAP3Ks MAP3K17和MAP3K18是MKK3的上游MAP3Ks,涉及ABA信号传导中的MAP3K17/18-MKK3-MPK1/2/7/14级联。据报道,MAP3K18在拟南芥中的过表达导致叶片早衰表型、ABA超敏气孔关闭和耐旱性。在本研究中,我们产生了过表达MAP3K17(35S:MAP3K17)及其激酶非活性形式(35S:MAP3K17KN)的转基因植物。35S:MAP3K17的抽薹早于WT,幼苗的鲜重较小,而35S:MAP3K17KN表现出相反的表型。这些结果表明,从营养生长到生殖生长的转变可以通过过表达MAP3K17及其激酶非活性形式来调节。此外,35S:MAP3K17在发芽后生长过程中对ABA的敏感性较低,而35S:MMAP3K17KN表现出相反的表型,这表明MAP3K17在对ABA的反应中起着负面作用。我们的工作提供了通过ABA诱导的MAPK级联的遗传操作来调节植物生长和发育的可能性,从而提高作物生长和生产力。
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引用次数: 11
An improved anther culture procedure for obtaining new commercial Mediterranean temperate japonica rice (Oryza sativa) genotypes. 用于获得新的商业地中海温带粳稻(Oryza sativa)基因型的改良花药培养程序。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0409A
C. López-Cristoffanini, X. Serrat, Eduardo Ramos-Fuentes, Isidre Hooghvorst, Roser Llaó, M. López-Carbonell, S. Nogués
Rice is one of the greatest calorie supply for the world population, especially since its production is almost entirely destined to direct human consumption and its demand will increase along with the world population. There are efforts worldwide to increase rice yields by obtaining new improved and stabilized rice lines. The rice anther culture, a fast and cheap technique, allows to obtain double haploid lines in less than one year. We report its application with an improved protocol in four Mediterranean japonica rice genotypes at F2 generation. We performed a screening test for cold-pretreatment at 5.0±0.1°C and concluded that the optimum duration was 9 days as it produced the higher rate of anther-derived callus induction. This revised protocol was successfully applied to the four genotypes, obtaining good results in all the procedure's steps. At the end, more than 100 of double haploid green plants were generated. Moreover, 9 lines obtained from the anther culture procedure showed good qualities for the Spanish market at the growing, farming and grain production level during the field assays. Therefore, we report an improved anther culture procedure for obtaining double haploid lines from temperate japonica rice genotypes showing high commercialization expectance.
大米是世界人口最大的卡路里供应之一,特别是因为它的生产几乎完全是为了直接供人类消费,它的需求将随着世界人口的增加而增加。全世界都在努力通过获得新的改良和稳定的水稻品系来提高水稻产量。水稻花药培养是一种快速而廉价的技术,可以在不到一年的时间内获得双单倍体系。本文报道了其改良方案在4个地中海粳稻基因型F2代的应用。我们进行了5.0±0.1°C冷预处理的筛选试验,得出最佳处理时间为9天,其花药愈伤组织诱导率较高。修订后的方案成功地应用于四种基因型,在所有步骤中都获得了良好的结果。最终获得100多株双单倍体绿色植株。此外,从花药培养过程中获得的9个品系在种植、耕作和粮食生产水平上都表现出良好的品质,可供西班牙市场使用。因此,我们报告了一种改进的花药培养程序,从具有高商业化预期的温带粳稻基因型中获得双单倍体系。
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引用次数: 7
AgarTrap Protocols on your Benchtop: Simple Methods for Agrobacterium-mediated Genetic Transformation of the Liverwort Marchantia polymorpha. 台式AgarTrap协议:农杆菌介导的多形地茅遗传转化的简单方法。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0312B
Shoko Tsuboyama, Y. Kodama
Agrobacterium-mediated genetic transformation is a powerful technique in plant biology. We recently developed a simplified Agrobacterium-mediated genetic transformation method for the liverwort Marchantia polymorpha, named AgarTrap (agar-utilized transformation with pouring solutions). AgarTrap is easy to perform; all procedures can be completed within a week using a single plate of solid medium, and basic operations involve simply pouring the appropriate solutions onto the solid medium. Thus far, we have developed three types of AgarTrap methods (S-AgarTrap, G-AgarTrap, and T-AgarTrap) using three different M. polymorpha tissues: sporelings, intact gemmalings, and mature thallus pieces, respectively. Each AgarTrap method can be used to transform tissues at high efficiency, thereby producing sufficient numbers of transformants for study. The ease and efficiency of these AgarTrap methods will likely prompt widespread molecular biological analyses of M. polymorpha. In this review, we describe the basic characteristics of the three AgarTrap methods and present the detailed protocols used in our laboratory.
农杆菌介导的遗传转化是植物生物学中一项强有力的技术。我们最近开发了一种简化的农杆菌介导的多态地衣遗传转化方法,命名为AgarTrap(琼脂利用转化与浇注溶液)。AgarTrap很容易执行;所有的步骤都可以在一周内用一块固体培养基完成,基本操作包括简单地将适当的溶液倒在固体培养基上。到目前为止,我们已经开发了三种类型的AgarTrap方法(S-AgarTrap, G-AgarTrap和T-AgarTrap),分别使用三种不同的多态m组织:孢子、完整的小苗和成熟的菌体片段。每种AgarTrap方法都可以高效地转化组织,从而产生足够数量的转化子用于研究。这些AgarTrap方法的简便和高效可能会促进多形霉分子生物学分析的广泛开展。在这篇综述中,我们描述了三种AgarTrap方法的基本特征,并介绍了我们实验室使用的详细方案。
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引用次数: 20
Efficient identification of NLR by using a genome-wide protein domain and motif survey program, Ex-DOMAIN. 使用全基因组蛋白质结构域和基序调查程序Ex domain有效鉴定NLR。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/plantbiotechnology.18.0418a
M. Narusaka, H. Yunokawa, Y. Narusaka
Genomic and amino acid sequences of organisms are freely available from various public databases. We designed a genome-wide survey program, named "Ex-DOMAIN" (exhaustive domain and motif annotator using InterProScan), of protein domains and motifs to aid in the identification and characterization of proteins by using the InterProScan sequence analysis application, which can display information and annotations of targeted proteins and genes, conserved protein domains and motifs, chromosomal locations, and structural diversities of target proteins. In this study, we indicated the disease resistance genes (proteins) that play an important role in defense against pathogens in Arabidopsis thaliana (thale cress) and Cucumis sativus (cucumber), by searches based on the conserved protein domains, NB-ARC (a nucleotide-binding adaptor shared by the apoptotic protease-activating factor-1, plant resistance proteins, and Caenorhabditis elegans death-4 protein) and C-terminal leucine-rich repeat (LRR), in the nucleotide-binding domain and LRR (NLR) proteins. Our findings suggest that this program will enable searches for various protein domains and motifs in all organisms.
生物体的基因组和氨基酸序列可以从各种公共数据库中免费获得。我们设计了一个名为“Ex-DOMAIN”(使用InterProScan的详尽结构域和基序注释器)的全基因组蛋白质结构域和基序调查程序,以通过使用InterProScan-序列分析应用程序帮助识别和表征蛋白质,该程序可以显示靶向蛋白质和基因、保守蛋白质结构域,染色体位置和靶蛋白的结构多样性。在本研究中,我们通过基于保守蛋白结构域的搜索,指出了在拟南芥(thale cress)和黄瓜(Cucumis sativus)中对病原体防御起重要作用的抗病基因(蛋白质),NB-ARC(一种由凋亡蛋白酶激活因子-1、植物抗性蛋白和秀丽隐杆线虫死亡-4蛋白共享的核苷酸结合接头)和核苷酸结合结构域和LRR(NLR)蛋白中的C-末端富含亮氨酸重复序列(LRR)。我们的发现表明,该程序将能够在所有生物体中搜索各种蛋白质结构域和基序。
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引用次数: 1
Identification and characterization of a novel sesquiterpene synthase, 4-amorphen-11-ol synthase, from Artemisia maritima. 一种新的倍半萜合酶,4-阿莫酚-11-醇合酶的鉴定和鉴定。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0324A
Paskorn Muangphrom, H. Seki, S. Matsumoto, M. Nishiwaki, E. O. Fukushima, T. Muranaka
Artemisinin, a sesquiterpene lactone exhibiting effective antimalarial activity, is produced by only Artemisia annua plant. A key step in artemisinin biosynthesis is the cyclization of farnesyl pyrophosphate (FPP) to amorpha-4,11-diene catalyzed by amorpha-4,11-diene synthase (AaADS). Intriguingly, several non-artemisinin-producing Artemisia plants also express genes highly homologous to AaADS. Our previous functional analysis of these homologous enzymes revealed that they catalyzed the synthesis of rare natural sesquiterpenoids. In this study, we analyzed the function of another putative sesquiterpene synthase highly homologous to AaADS from A. maritima. Unlike AaADS, in vivo enzymatic assay showed that this enzyme cyclized FPP to 4-amorphen-11-ol, a precursor of several gastroprotective agents. The discovery of 4-amorphen-11-ol synthase (AmAOS) and the successful de novo production of 4-amorphen-11-ol in engineered yeast demonstrated herein provides insights into the methods used to enhance its production for future application.
青蒿素是一种倍半萜内酯,具有有效的抗疟活性,仅由青蒿属植物产生。青蒿素生物合成的一个关键步骤是在阿莫酚-4,11-二烯合酶(AaADS)的催化下,法尼焦磷酸酯(FPP)环化为阿莫酚-4,11-二烯。有趣的是,一些不产生青蒿素的青蒿素植物也表达与AaADS高度同源的基因。我们之前对这些同源酶的功能分析表明,它们催化了罕见的天然倍半萜的合成。在这项研究中,我们分析了另一种推测的倍半萜合成酶的功能,该合成酶与来自A.maritima的AaADS高度同源。与AaADS不同,体内酶测定显示,这种酶将FPP环化为4-阿莫酚-11-醇,这是几种胃保护剂的前体。本文证明的4-阿莫酚-11-醇合酶(AmAOS)的发现和在工程酵母中成功从头生产4-阿莫芬-11-醇为未来应用提供了对用于增强其生产的方法的深入了解。
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引用次数: 4
Comparative analysis of CYP716A subfamily enzymes for the heterologous production of C-28 oxidized triterpenoids in transgenic yeast. 转基因酵母中异源产生C-28氧化三萜的CYP716A亚家族酶的比较分析。
IF 1.6 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2018-06-25 DOI: 10.5511/PLANTBIOTECHNOLOGY.18.0416A
Hayato Suzuki, E. O. Fukushima, N. Umemoto, K. Ohyama, H. Seki, T. Muranaka
Several enzymes of the CYP716A subfamily have been reported to be involved in triterpenoid biosynthesis. Members of this subfamily oxidize various positions along the triterpenoid backbone and the majority of them catalyze a three-step oxidation at the C-28 position. Interestingly, C-28 oxidation is a common feature in oleanolic acid, ursolic acid, and betulinic acid, which are widely distributed in plants and exhibit important biological activities. In this work, three additional CYP716A enzymes isolated from olive, sugar beet, and coffee, were characterized as multifunctional C-28 oxidases. Semi-quantitative comparisons of in vivo catalytic activity were made against the previously characterized enzymes CYP716A12, CYP716A15, and CYP716A52v2. When heterologously expressed in yeast, the isolated enzymes differed in both catalytic activity and substrate specificity. This study indicates that the screening of enzymes from different plants could be a useful means of identifying enzymes with enhanced catalytic activity and desired substrate specificity. Furthermore, we show that "naturally-evolved" enzymes can be useful in the heterologous production of pharmacologically and industrially important triterpenoids.
据报道,CYP716A亚家族的几种酶参与了三萜的生物合成。这个亚家族的成员沿着三萜骨架氧化不同的位置,其中大多数在C-28位置催化三步氧化。有趣的是,C-28氧化是齐墩果酸、熊果酸和白桦酸的共同特征,它们广泛分布于植物中并具有重要的生物活性。在这项工作中,从橄榄,甜菜和咖啡中分离出另外三种CYP716A酶,被表征为多功能C-28氧化酶。与先前表征的酶CYP716A12、CYP716A15和CYP716A52v2进行了体内催化活性的半定量比较。当在酵母中异种表达时,分离的酶在催化活性和底物特异性上都有所不同。该研究表明,从不同植物中筛选酶可能是鉴定具有增强催化活性和所需底物特异性的酶的有效手段。此外,我们发现“自然进化”的酶可用于异源生产药理学和工业上重要的三萜。
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引用次数: 18
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Plant Biotechnology
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