• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.135-153
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• 2015

The aromatic amino acids, which are composed of $\small{L}$-phenylalanine, $\small{L}$-tyrosine and $\small{L}$-tryptophan, are general components of protein synthesis as well as precursors for a wide range of secondary metabolites. These aromatic amino acids-derived compounds play important roles as ingredients of diverse phenolics including pigments and cell walls, and hormones like auxin and salicylic acid in plants. Moreover, they also serve as the natural products of alkaloids and glucosinolates, which have a high potential to promote human health and nutrition. The biosynthetic pathways of aromatic amino acids share a chorismate, the common intermediate, which is originated from shikimate pathway. Then, tryptophan is synthesized via anthranilate and the other phenylalanine and tyrosine are synthesized via prephenate, as intermediates. This review reports recent studies about all the enzymatic steps involved in aromatic amino acid biosynthetic pathways and their gene regulation on transcriptional/post-transcriptional levels. Furthermore, results of metabolic engineering are introduced as efforts to improve the production of the aromatic amino acids-derived secondary metabolites in plants.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.506-514
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• 2020

4-Hydroxybenzyl alcohol (4-HB alcohol) is one of the major active components of Gastrodia elata Blume, with beneficial effects on neurological disorders such as headache, convulsive behavior, and dizziness. Here, we developed a metabolically engineered Corynebacterium glutamicum strain able to produce 4-HB alcohol from 4-hydroxybenzoate (4-HBA). First, the strain APS963 was obtained from the APS809 strain via the insertion of aroK from Methanocaldococcus jannaschii into the NCgl2922-deleted locus. As carboxylic acid reductase from Nocardia iowensis catalyzes the reduction of 4HBA to 4-hydroxybenzaldehyde (4-HB aldehyde), we then introduced a codon-optimized car gene into the genome of APS963, generating the GAS177 strain. Then, we deleted creG coding for a putative short-chain dehydrogenase and inserted ubiCpr encoding a product-resistant chorismate-pyruvate lyase into the pcaHG-deleted locus. The resulting engineered GAS355 strain accumulated 2.3 g/l 4-HB alcohol with 0.32 g/l 4-HBA and 0.3 g/l 4-HB aldehyde as byproducts from 8% glucose after 48 h of culture.

• Journal of Microbiology and Biotechnology
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• v.25 no.1
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• pp.66-73
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• 2015

The pabS gene of Agaricus bisporus 02 encoding a putative PABA synthase was cloned, and then the recombinant protein was expressed in Escherichia coli BL21 under the control of the T7 promoter. The enzyme with an N-terminal GST tag or His tag, designated GST-AbADCS or His-AbADCS, was purified with glutathione Sepharose 4B or Ni Sepharose 6 Fast Flow. The enzyme was an aminodeoxychorismate synthase, and it was necessary to add with an aminodeoxychorismate lyase for synthesizing PABA. AbADCS has maximum activity at a temperature of approximately 25℃ and pH 8.0. Magnesium or manganese ions were necessary for the enzymatic activity. The Michaelis-Menten constant for chorismate was 0.12 mM, and 2.55 mM for glutamine. H₂O₂ did distinct damage on the activity of the enzyme, which could be slightly recovered by Hsp20. Sulfydryl reagents could remarkably promote its activity, suggesting that cysteine residues are essential for catalytic function.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.169-176
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• 2008

Salicylic acid(SA) is a phytohormone that is related to plant defense mechanism. The SA accumulation is triggered by abiotic and biotic stresses. SA acts as a signal molecular compound mediating systemic acquired resistance and hypersensitive response in plant. Although the role of SA has been studied extensively, an understanding of the SA regulatory mechanism is still lacking in plants. In order to comprehend SA regulatory mechanism, we have been transformed with a SID2 promoter:GUS::LUC fusion construct into siz1-2 mutant and wild plant(Col-0). SIZ1 encodes SUMO E3 ligase and negatively regulates SA accumulation in plants. SID2(SALICYLIC ACID INDUCTION DEFICIENT2) is a crucial enzyme of SA biosynthesis. The Arabidopsis SID2 gene encodes isochorismate synthase(ICS) that controls SA level by conversion of chorismate to isochorismate. We compared the regulation of SID2 in wild-type and siz1-2 transgenic plants that express SID2 promoter:GUS::LUC constructs respectively. The expressions of $\beta$-GLUCURONIDASE and LUCIFERASE were higher in siz 1-2 transgenic plant without any stress treatment. SID2 promoter:GUS::LUC/siz1-2 transgenic plant will be used as a starting material for isolation of siz1-2 suppressor mutants and genes involved in SA-mediated stress signaling pathway.