Intercellular Localization of Cysteine Synthase and Alliinase in Bundle Sheaths of Allium Plants

Yamazaki, Mami; Sugiyama, Mitsuyo; Saito, Kazuki

doi:10.5511/plantbiotechnology.19.7

Cited by 16 publications

(7 citation statements)

References 11 publications

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“…Interestingly, bulk XAS spectra of epidermal peels did not show corresponding changes on rubbing (not illustrated), even when rubbing was quite extensive (causing essentially 100% cell breakage), although the features attributable to sulfoxide content were comparable to and slightly more intense than the onion spectra depicted in Figure 4. Presumably this lack of reactivity is due to a lack of alliinase in the epidermal cell layers, which is consistent with earlier work indicating preferential localization of alliinase in bundle sheath cells (30).…”

Section: Resultssupporting

confidence: 91%

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Localizing the Chemical Forms of Sulfur in Vivo Using X-ray Fluorescence Spectroscopic Imaging: Application to Onion (Allium cepa) Tissues

et al. 2009

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Sulfur has a particularly rich biochemistry and fills a number of important roles in biology. In situ information on sulfur biochemistry is generally difficult to obtain because of a lack of biophysical techniques that have sufficient sensitivity to molecular form. We have recently reported that sulfur K-edge X-ray absorption spectroscopy can be used as a direct probe of the sulfur biochemistry of living mammalian cells [Gnida, M., et al. (2007) Biochemistry 46, 14735-14741]. Here we report an extension of this work and develop sulfur K-edge X-ray fluorescence spectroscopic imaging as an in vivo probe of sulfur metabolism in living cells. For this work, we have chosen onion (Allium cepa) as a tractable model system with well-developed sulfur biochemistry and present evidence of the localization of a number of different chemical forms. X-ray absorption spectroscopy of onion sections showed increased levels of lachrymatory factor (LF) and thiosulfinate and decreased levels of sulfoxide (LF precursor) following cell breakage. In intact cells, X-ray fluorescence spectroscopic imaging showed elevated levels of sulfoxides in the cytosol and elevated levels of reduced sulfur in the central transport vessels and bundle sheath cells.

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Section: Resultssupporting

confidence: 91%

“…This study indicated that the sulfoxide precursor and the alliinase enzyme are located in the cell vacuole and cytosol, respectively. Yamazaki et al (30) later used immunofluorescent labeling to demonstrate that alliinase is preferentially localized in bundle sheath cells.…”

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confidence: 99%

Localizing the Chemical Forms of Sulfur in Vivo Using X-ray Fluorescence Spectroscopic Imaging: Application to Onion (Allium cepa) Tissues

et al. 2009

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“…Alliinase has been purified from garlic, onion, and other plants of the genus Allium (Schwimmer and Mazelis, 1963, Mazelis and Crews, 1968, Tobkin and Mazelis, 1979, Nock and Mazelis, 1987, Landshuter et al, 1994, Lohmuller et al, 1994, Rabinkov et al, 1994, Manabe et al, 1998). The enzymatic production of allicin occurs in garlic, because of the injury of the plant tissue that enables interaction of the enzyme in vacuoles with alliin gathered in the cytosol (Yamazaki et al, 2002); hence allicin production has been discussed as a defense mechanism of the plant against microbial infection or insect attack (Slusarenko et al, 2008). Allicin generated from alliinase which was purified from garlic, has been studied for fungicidal activity against Magnaporthe grisea (Fry et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Purification and characterisation of alliinase produced by Cupriavidus necator and its application for generation of cytotoxic agent: Allicin

Chhabria

Desai

2018

Saudi Journal of Biological Sciences

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Allicin, an extremely active constituent of freshly crushed garlic, is produced upon reaction of alliin with the enzyme alliinase (EC 4.4.1.4). A bacterium Cupriavidus necator with the ability of alliinase production was isolated from a soil sample and was identified by morphological, biochemical and 16S rRNA sequence. Alliinase production was optimised and it was further purified to apparent homogeneity with 103-fold purification and specific activity of 209 U/mg of protein by using DEAE Cellulose and Sephadex G-100 chromatography. The enzyme is a homodimer of molecular weight 110 kDa with two subunits of molecular weight 55 kDa each. The optimum activity of the purified enzyme was found at pH 7 and the optimum temperature was 35 °C. The enzyme exhibited maximum reaction rate (Vmax) at 74.65 U/mg and Michaelis–Menten constant (Km) was determined to be 0.83 mM when alliin was used as a substrate. The cytotoxic activity of in-situ generated allicin using purified alliinase and alliin was assessed on MIA PaCa-2 cell line using MTT assay and Acridine orange–ethidium bromide staining. This approach of in-situ allicin generation suggests a novel therapeutic strategy wherein alliin and alliinase work together synergistically to produce cytotoxic agent allicin.

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“…2 In whole intact garlic cloves, alliinase accumulated in bundle sheath cells converts (+)-alliin to allicin after garlic is crushed or sliced. 3 The resulting allicin appears to be chemically unstable, as it spontaneously changes into various sulfide-containing compounds. 2,4 These products exhibit antibacterial, cholesterol-lowering, anti-inflammatory, antioxidant, and anticancer activity; therefore, they have healthpromoting effects for human.…”

Section: Introductionmentioning

confidence: 99%

One-Pot Synthesis of Useful S-Substituted-l-cysteine Sulfoxides Using Genetically Engineered Escherichia coli

Mizutani,

Hara,

Takeuchi

et al. 2024

J. Agric. Food Chem.

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S-Substituted-L-cysteine sulfoxides are valuable compounds that are contained in plants. Particularly, (+)-alliin and its degraded products have gained significant attention because of their human health benefits. However, (+)-alliin production has been limited to extraction from plants and chemical synthesis; both methods have drawbacks in terms of stability and safety. Here, we proposed the enzymatic cascade reaction for synthesizing (+)-alliin from readily available substrates. To achieve a one-pot (+)-alliin production, we constructed Escherichia coli coexpressing the genes encoding tryptophan synthase from Aeromonas hydrophila ssp. hydrophila NBRC 3820 and L-isoleucine hydroxylase from Bacillus thuringiensis 2e2 for the biocatalyst. Deletion of tryptophanase gene in E. coli increased the yield about 2-fold. Under optimized conditions, (+)-alliin accumulation reached 110 mM, which is the highest productivity thus far. Moreover, natural and unnatural S-substituted-L-cysteine sulfoxides were synthesized by applying various thiols to the cascade reaction. These results indicate that the developed bioprocess would enable the supply of diverse Ssubstituted-L-cysteine sulfoxides.

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Intercellular Localization of Cysteine Synthase and Alliinase in Bundle Sheaths of Allium Plants

Cited by 16 publications

References 11 publications

Localizing the Chemical Forms of Sulfur in Vivo Using X-ray Fluorescence Spectroscopic Imaging: Application to Onion (Allium cepa) Tissues

Localizing the Chemical Forms of Sulfur in Vivo Using X-ray Fluorescence Spectroscopic Imaging: Application to Onion (Allium cepa) Tissues

Purification and characterisation of alliinase produced by Cupriavidus necator and its application for generation of cytotoxic agent: Allicin

One-Pot Synthesis of Useful S-Substituted-l-cysteine Sulfoxides Using Genetically Engineered Escherichia coli

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