[논문]Biosynthetic Pathway of Indole-3-Acetic Acid in Basidiomycetous Yeast Rhodosporidiobolus fluvialis

Bunsangiam, Sakaoduoen; Sakpuntoon, Varunya; Srisuk, Nantana; Ohashi, Takao; Fujiyama, Kazuhito; Limtong, Savitree

doi:10.1080/12298093.2019.1638672

[국내논문] Biosynthetic Pathway of Indole-3-Acetic Acid in Basidiomycetous Yeast Rhodosporidiobolus fluvialis 원문보기

Mycobiology, v.47 no.3, 2019년, pp.292 - 300

Bunsangiam, Sakaoduoen (Department of Microbiology, Faculty of Science, Kasetsart University) , Sakpuntoon, Varunya (Department of Microbiology, Faculty of Science, Kasetsart University) , Srisuk, Nantana (Department of Microbiology, Faculty of Science, Kasetsart University) , Ohashi, Takao (International Center for Biotechnology, Osaka University) , Fujiyama, Kazuhito (International Center for Biotechnology, Osaka University) , Limtong, Savitree (Department of Microbiology, Faculty of Science, Kasetsart University)

Abstract ▼ AI-Helper

IAA biosynthetic pathways in a basidiomycetous yeast, Rhodosporidiobolus fluvialis DMKU-CP293, were investigated. The yeast strain showed tryptophan (Trp)-dependent IAA biosynthesis when grown in tryptophan supplemented mineral salt medium. Gas chromatography-mass spectrometry was used to further identify the pathway intermediates of Trpdependent IAA biosynthesis. The results indicated that the main intermediates produced by R. fluvialis DMKU-CP293 were tryptamine (TAM), indole-3-acetic acid (IAA), and tryptophol (TOL), whereas indole-3-pyruvic acid (IPA) was not found. However, supplementation of IPA to the culture medium resulted in IAA peak detection by high-performance liquid chromatography analysis of the culture supernatant. Key enzymes of three IAA biosynthetic routes, i.e., IPA, IAM and TAM were investigated to clarify the IAA biosynthetic pathways of R. fluvialis DMKU-CP293. Results indicated that the activities of tryptophan aminotransferase, tryptophan 2-monooxygenase, and tryptophan decarboxylase were observed in cell crude extract. Overall results suggested that IAA biosynthetic in this yeast strain mainly occurred via the IPA route. Nevertheless, IAM and TAM pathway might be involved in R. fluvialis DMKU-CP293.

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문제 정의

fluvialis on IAA biosynthesis has not yet been reported. Therefore, the aim of this study was to focus on the clarification of the IAA biosynthetic pathways in R. fluvialis.

제안 방법

To investigate whether R. fluvialis DMKU-CP293 employed Trp-dependent or Trp-independent IAA biosynthesis, culture media with and without tryptophan supplements were used. The inoculum was transferred into a 250 ml Erlenmeyer flask containing 50 ml mineral salt (MS) medium (gram per liter: KH₂PO₄, 1; K2HPO₄, 1; NaCl, 1; MgSO₄・7H₂。, 4; (NH₄)2 SO₄, 4; Na3C₆H₅O₇・7H₂。, 0.
The experiments were carried out by feeding the yeast with possible IAA precursors, such as IPA, IAM, TAM, and IAN. The precultured inoculum of the yeast strains was resuspended in MS medium from which 0.
fluvialis is still required. To investigate tryptophan dependency for IAA biosynthesis of R. fluvialis DMKU-CP293, the culture supernatant of yeast culture grown in MS medium, with or without 0.1% L- tryptophan, was analyzed by HPLC. After 5 days of incubation, yeast stain produced amount (363.
Finding of tryptophan aminotransferase activity in cell crude extract indicated that the IPA route is probably one of the IAA biosynthetic pathways in this yeast. To determine the key enzyme of the IAM pathway, the active conversion of tryptophan to IAM was investigated by an in vitro assay of the tryptophan 2-monooxygenase activity and IAM was measured by the GC-MS. The formation of IAM was detected to indicate that tryptophan 2-monooxygenase activity present in R.
To assure the validity of our results described above, feeding experiments with different types of IAA intermediates were carried out and IAA was detected by HPLC analysis. IPA, IAM, IAN, and TAM were individually added as a precursor.

성능/효과

fluvialis DMKU-CP293 may use for IAA biosynthesis by determining the pathway intermediates produced by yeast. The results indicated that TAM and IAM, but not IPA, were found in the culture medium of R. fluvialis DMKU-CP293. TOL, which is an aromatic alcohol and is a product of enzymatic reduction of IAAld, was also detected.
To investigate the involvement of the IAN pathway with IAA biosynthesis in this yeast nitrile hydratase and nitrilase activities were assayed but none was found (Table 1). Therefore, the results presented in this study provided evidence that R. fluvialis DMKU-CP293 may use IPA, IAM, and TAM pathways for IAA biosynthesis.

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[국내논문] Biosynthetic Pathway of Indole-3-Acetic Acid in Basidiomycetous Yeast Rhodosporidiobolus fluvialis 원문보기

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[국내논문] Biosynthetic Pathway of Indole-3-Acetic Acid in Basidiomycetous Yeast Rhodosporidiobolus fluvialis 원문보기

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