• Applied Biological Chemistry
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• 제33권1호
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• pp.68-72
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• 1990

저온에서 일어나는 식물생육의 억제와 식물생장조절물질의 하나인 IAA 수준의 조절을 알아보기 위하여 암실재배한 백색 완두유묘를 재료로 저온처리에 따른 생육과 IAA 수준의 변화를 조사하였다. $25^{\circ}C$에서 성장한 백색 완두유묘는 $5^{\circ}C$의 저온에 접하면 생장이 거의 정지하였다. 유리 IAA 수준은 생체중 기준 상온묘의 17.6 ng/g에서 3일간 저온처리시에 4.9 ng/g으로, 총 IAA 수준은 113.2 ng/g에서 60.8 ng/g으로 현저하게 감소하였다. IAA의 전구체인 트립토판 수준은 생체중 기준 상온묘의 583 nmol/g에서 저온처리시에 414 nmol/g으로 약 30 % 감소하였으며, 시킴산경로에 의하여 트립토판과 함께 생합성되는 페닐알라닌과 티로신 수준도 감소하였다. 이 결과는 저온에서 IAA의 전구체인 트립토판 생합성의 억제를 통하여 IAA 수준을 일부 하향조절할 수 있을 것임을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제20권9호
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• pp.1259-1265
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• 2010

The filamentous cyanobacterium Arthrospira platensis strain MMG-9 was isolated from a rice field. The ability of this strain to synthesize the bioactive compound indole-3-acetic acid (IAA) was demonstrated. IAA was extracted from the culture of A. platensis strain MMG-9 and its identity was confirmed by thin-layer chromatography (TLC) as well as by high-performance liquid chromatography (HPLC). The IAA precursor L-tryptophan was required for IAA biosynthesis. Released IAA increased with the increase of the initial concentration of L-tryptophan in the medium and with the incubation time. A. platensis strain MMG-9 accumulated more IAA than it released into the medium. The bioactivity of the secreted IAA was shown by its effect on the formation of roots by Pisum sativum. There was a significant positive effect of the supernatant of cultures of A. platensis strain MMG-9 on the number of lateral roots of P. sativum, whereas a negative effect on root length was observed.

• 한국육종학회지
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• 제43권1호
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• pp.1-13
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• 2011

Low temperature stress is one of the major negative factors affecting vegetative and reproductive growth of rice. To better understand responses of rice plants to low temperature we analyzed transcriptome expression patterns in glumous flower of cold-tolerant japonica rice variety, Stejaree45, and cold-susceptible variety, HR19621-AC6 at booting stage under cold water irrigation. A total of 2,411 probes were differentially expressed by low temperature in glumous flowers of the two varieties. Some important genes involved in hormone biosynthesis showed variety-specific regulation. Expression of GA20ox3 and GA2ox, among the genes involved in GA biosynthesis, was regulated differentially in the two varieties. Among the genes involved in IAA biosynthesis, YUCCA1 and TAA1:1 showed variety-specific regulation. Among the genes involved in cytokinin biosynthsis and signaling, expression of LOG, HK1 and HK3 was significantly down-regulated only in the cold-susceptible variety. Among the genes involved in ABA biosynthesis, NSY and AAO3 were down-regulated only in the cold-tolerant variety. In general, genes involved in GA, IAA and cytokinin biosynthesis responded to cold temperature in such a way that capacity of those bioactive hormones is maintained at relatively higher levels under cold temperature in the cold-tolerant variety, which can help minimize cold stress imposed to developing reproductive organs in the cold-tolerant variety.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1726-1736
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• 2013

Biosynthesis of indole-3-acetic acid (IAA) via the indole-3-pyruvic acid pathway involves three kinds of enzymes; aminotransferase encoded by aspC, indole-3-pyruvic acid decarboxylase encoded by ipdC, and indole-3-acetic acid dehydrogenase encoded by iad1. The ipdC from Enterobacter cloacae ATCC 13047, aspC from Escherichia coli, and iad1 from Ustilago maydis were cloned and expressed under the control of the tac and sod promoters in E. coli. According to SDS-PAGE and enzyme activity, IpdC and Iad1 showed good expression under the control of $P_{tac}$, whereas AspC was efficiently expressed by $P_{sod}$ originating from Corynebacterium glutamicum. The activities of IpdC, AspC, and Iad1 from the crude extracts of recombinant E. coli Top 10 were 215.6, 5.7, and 272.1 nmol/min/mg-protein, respectively. The recombinant E. coli $DH5{\alpha}$ expressing IpdC, AspC, and Iad1 produced about 1.1 g/l of IAA and 0.13 g/l of tryptophol (TOL) after 48 h of cultivation in LB medium with 2 g/l tryptophan. To improve IAA production, a tnaA gene mediating indole formation from tryptophan was deleted. As a result, E. coli IAA68 with expression of the three genes produced 1.8 g/l of IAA, which is a 1.6-fold increase compared with wild-type $DH5{\alpha}$ harboring the same plasmids. Moreover, the complete conversion of tryptophan to IAA was achieved by E. coli IAA68. Finally, E. coli IAA68 produced 3.0 g/l of IAA after 24 h cultivation in LB medium supplemented with 4 g/l of tryptophan.

• 생명과학회지
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• 제19권8호
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• pp.1139-1144
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• 2009

옥수수 뿌리에 BL을 처리하면 양성굴중성 반응이 촉진되고, ethylene 생성도 증가한다는 것이 알려져 있다. BL에 의해 유도된 굴중성 반응과 ethylene 생성과의 관계를 조사하였다. ethylene 생성 억제제인 $10^{-4}$ M AVG를 처리하면 ethylene 생성은 90% 이상 억제되었으나, 굴중성 반응은 13% 정도 억제되었다. AVG를 BL과 함께 처리한 뿌리는 ethylene 생성은 약 60% 억제되었으나 굴중성 반응은 대조구 보다 증가하였다. 다른 ethylene 생성 억제제인 cobalt ion을 처리하면 ethylene 생성은 약 10% 정도 억제되었으나 굴중성 반응은 억제되지 않았다. BL과 cobalt ion을 함께 처리한 뿌리는 ethylene 생성이 억제되었으나 굴중성 반응은 증가되었다. 이러한 BL의 효과가 auxin transport와 관계가 있는지 알아보기 위하여 auxin transport inhibitor인 TIBA를 처리하였다. $10^{-5}$ M TIBA와 BL과 TIBA를 함께 처리한 경우(BL+TIBA), ethylene 생성은 각각 96%, 132% 증가하였으나 굴중성 반응은 모두 일어나지 않았다. 또한, BL, TIBA 그리고 IAA를 함께 처리한 뿌리 (BL+TIBA+IAA)는 음성굴중성 반응을 나타냈으나 뿌리 생장은 오히려 증가시켰으며, 이는 수평으로 있는 뿌리에서 IAA가 아랫면으로 transport 되지 못하고 윗면에 축적된 것을 의미한다. 이러한 결과는 BL이 뿌리 내에 존재하는 IAA의 차등분포에 영향을 주어 양성굴중성 반응을 촉진할 가능성을 제시한다.

• Molecules and Cells
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• 제41권12호
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• pp.1033-1044
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• 2018

As sessile organisms, plants have evolved to adjust their growth and development to environmental changes. It has been well documented that the crosstalk between different plant hormones plays important roles in the coordination of growth and development of the plant. Here, we describe a novel recessive mutant, mildly insensitive to ethylene (mine), which displayed insensitivity to the ethylene precursor, ACC (1-aminocyclopropane-1-carboxylic acid), in the root under the dark-grown conditions. By contrast, mine roots exhibited a normal growth response to exogenous IAA (indole-3-acetic acid). Thus, it appears that the growth responses of mine to ACC and IAA resemble those of weak ethylene insensitive (wei) mutants. To understand the molecular events underlying the crosstalk between ethylene and auxin in the root, we identified the MINE locus and found that the MINE gene encodes the pyridoxine 5′-phosphate (PNP)/pyridoxamine 5′-phosphate (PMP) oxidase, PDX3. Our results revealed that MINE/PDX3 likely plays a role in the conversion of the auxin precursor tryptophan to indole-3-pyruvic acid in the auxin biosynthesis pathway, in which TAA1 (TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1) and its related genes (TRYPTOPHAN AMINOTRANSFERASE RELATED 1 and 2; TAR1 and TAR2) are involved. Considering that TAA1 and TARs belong to a subgroup of PLP (pyridoxal-5′-phosphate)-dependent enzymes, we propose that PLP produced by MINE/PDX3 acts as a cofactor in TAA1/TAR-dependent auxin biosynthesis induced by ethylene, which in turn influences the crosstalk between ethylene and auxin in the Arabidopsis root.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1998년도 한국생물과학협회 학술발표대회
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• pp.365.1-365
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• 1998

No Abstract, See Full Text

• 한국미생물·생명공학회지
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• 제47권2호
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• pp.242-249
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• 2019

Biosynthesis of indole-3-acetate (IAA) from L-tryptophan via indole-3-pyruvate pathway requires three enzymes including aminotransferase, indole-3-pyruvate decarboxylase, and indole-3-acetate dehydrogenase. To establish a bio-based production of IAA, the aspC, ipdC, and iad1 from Escherichia coli, Enterobacter cloacae, and Ustilago maydis, respectively, were expressed under control of the tac, ilvC, and sod promoters in C. glutamicum. Cells harboring ipdC produced tryptophol, indicating that the ipdC product is functional in this host. Analyses of SDS-PAGE and enzyme activity revealed that genes encoding AspC and Iad1 were efficiently expressed from the sod promoter, and their enzyme activities were 5.8 and 168.5 nmol/min/mg-protein, respectively. The final resulting strain expressing aspC, ipdC, and iad1 produced 2.3 g/l and 7.3 g/l of IAA from 10 g/l L-tryptophan, respectively, in flask cultures and a 5-L bioreactor.

• Journal of Microbiology and Biotechnology
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• 제18권7호
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• pp.1235-1244
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• 2008

Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.

• Applied Biological Chemistry
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• 제28권3호
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• pp.187-195
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• 1985

당근에서 cytokinin, auxin 또는 GA에 의해 조절되는 polypeptide 및 단백질들을 동정하기 위하여, kinetin, BA, IAA, NAA 혹은 $GA_3$가 각각 $10^{-4},\;10^{-5},\;10^{-6}M$인 배지에서 suspension culture 한 당근 callus들의 추출물의 전기영동양상을 조사하였다. 생장조절제를 처리하지 않은 callus에서 15개 polypeptide band가 관찰되었는데, 이들의 분자량은 각각 $18._4,\;20._2,\;24._0,\;34._9,\;35._7,\;37._4,\;40._3,\;42._2,\;44._1,\;44._4,\;49._3,\;55._0,\;56._6,\;58._1$ 그리고 $59._9KD$였다. Kinetin, BA, IAA, NAA 그리고 $GA_3$에 의하여 합성이 촉진되는 것으로 보이는 polypeptide band는 각각 2개, 6개, 1개, 2개 그리고 4개였고, 합성이 억제되는 것으로 보이는 것은 각각 5개, 3개, 4개, 3개, 그리고 2개였다. 분자량 $40._3KD$ 및 $42._2KD$의 polypeptide들은 cytokinin류 생장조절제에 의하여 합성이 촉진되는 것으로 나타났고, $44._1KD$의 것은 auxin류에 의하여 합성이 촉진되며 $37._4,\;44._4,\;56._6KD$의 것은 auxin류에 의하여 합성이 억제되는 것으로 나다났다. Kinetin, BA, IAA, NAA 및 $GA_3$처리에서 상대 이동도가 0.56, 0.84, 0.92인 단백질 band들이 증가된 것이 관찰되었으나, 각 생장조절제들에 특유한 단백질은 동정할 수 없었다.