Folate analysis in complex food matrices: Use of a recombinant Arabidopsis γ-glutamyl hydrolase for folate deglutamylation

Ramos-Parra, Perla A.; Urrea-López, Rafael; Garza, Rocío I. Díaz de la

doi:10.1016/j.foodres.2013.06.026

Cited by 25 publications

(23 citation statements)

References 32 publications

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“…Folates were extracted and quantified as previously described (Ramos‐Parra et al ., ) with some modifications. Seeds (0.5 g) were ground in liquid nitrogen, and the extract was treated for 2 h with protease (Sigma, St. Louis, MO).…”

Section: Methodsmentioning

confidence: 99%

Metabolic engineering of folate and its precursors in Mexican common bean (Phaseolus vulgaris L.)

Rivera

García‐Salinas

Aragão

et al. 2016

Plant Biotechnology Journal

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SummaryFolate (vitamin B9) deficiency causes several health problems globally. However, folate biofortification of major staple crops is one alternative that can be used to improve vitamin intakes in populations at risk. We increased the folate levels in common bean by engineering the pteridine branch required for their biosynthesis. GTP cyclohydrolase I from Arabidopsis (AtGchI) was stably introduced into three common bean Pinto cultivars by particle bombardment. Seed‐specific overexpression of AtGCHI caused significant increases of up to 150‐fold in biosynthetic pteridines in the transformed lines. The pteridine boost enhanced folate levels in raw desiccated seeds by up to threefold (325 μg in a 100 g portion), which would represent 81% of the adult recommended daily allowance. Unexpectedly, the engineering also triggered a general increase in PABA levels, the other folate precursor. This was not observed in previous engineering studies and was probably caused by a feedforward mechanism that remains to be elucidated. Results from this work also show that common bean grains accumulate considerable amounts of oxidized pteridines that might represent products of folate degradation in desiccating seeds. Our study uncovers a probable different regulation of folate homoeostasis in these legume grains than that observed in other engineering works. Legumes are good sources of folates, and this work shows that they can be engineered to accumulate even greater amounts of folate that, when consumed, can improve folate status. Biofortification of common bean with folates and other micronutrients represents a promising strategy to improve the nutritional status of populations around the world.

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Section: Methodsmentioning

confidence: 99%

Metabolic engineering of folate and its precursors in Mexican common bean (Phaseolus vulgaris L.)

Rivera

García‐Salinas

Aragão

et al. 2016

Plant Biotechnology Journal

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“…The cells were disrupted using a FastPrep Instrument (MP Biomedicals), boiled for 10 min, and centrifuged (16,000× g, 10 min, 4 • C). The supernatants were collected and treated with AtGGH2, as described by Ramos-Parra et al [39]. Folates were purified by affinity chromatography and analyzed by UPLC-coupled electrochemical detector (CoulArray Model 5600A; ESA, Chelmsford, MA) as described previously [40].…”

Section: Folate Profilingmentioning

confidence: 99%

In Vivo Rate of Formaldehyde Condensation with Tetrahydrofolate

Noor

Ramos-Parra

et al. 2020

Metabolites

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Formaldehyde is a highly reactive compound that participates in multiple spontaneous reactions, but these are mostly deleterious and damage cellular components. In contrast, the spontaneous condensation of formaldehyde with tetrahydrofolate (THF) has been proposed to contribute to the assimilation of this intermediate during growth on C1 carbon sources such as methanol. However, the in vivo rate of this condensation reaction is unknown and its possible contribution to growth remains elusive. Here, we used microbial platforms to assess the rate of this condensation in the cellular environment. We constructed Escherichia coli strains lacking the enzymes that naturally produce 5,10-methylene-THF. These strains were able to grow on minimal medium only when equipped with a sarcosine (N-methyl-glycine) oxidation pathway that sustained a high cellular concentration of formaldehyde, which spontaneously reacts with THF to produce 5,10-methylene-THF. We used flux balance analysis to derive the rate of the spontaneous condensation from the observed growth rate. According to this, we calculated that a microorganism obtaining its entire biomass via the spontaneous condensation of formaldehyde with THF would have a doubling time of more than three weeks. Hence, this spontaneous reaction is unlikely to serve as an effective route for formaldehyde assimilation.

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“…An earlier study found pentaglutamyl and heptaglutamyl folate are the major forms in black beans while hexaglutamyl folate is the major form in the tomato fruit; however, the polyglutamyl folate forms in the alfalfa sprouts shift to short-chain forms such as monoglutamyl and tetraglutamyl forms (23). The authors have realized that the polyglutamyl folate of alfalfa shifting towards short chains could be ascribed to the action of GGH activity during extraction.…”

Section: Pre-boiling Time Optimizationmentioning

confidence: 99%

“…There are several studies about quantitative polyglutamyl folate profiles in vegetables (17,(20)(21)(22)(23)(24)(25)(26). An earlier study found pentaglutamyl and heptaglutamyl folate are the major forms in black beans while hexaglutamyl folate is the major form in the tomato fruit; however, the polyglutamyl folate forms in the alfalfa sprouts shift to short-chain forms such as monoglutamyl and tetraglutamyl forms (23).…”

Section: Pre-boiling Time Optimizationmentioning

confidence: 99%

Quantification of Total Folate, Folate Species and Polyglutamyl Folate Distribution in Winged Beans (<i>Psophocarus tetragonolobus</i> (L) DC) from Different Cultivars and Growth Stages by Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry

Luo

Duan

Zou

et al. 2017

J Nutr Sci Vitaminol

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Summary Winged beans are an important natural source of some micronutrients. This paper presents the first complete characterization of folate derivatives including polyglutamyl 5-methyltetrahydrofolate (5-CH3-H4PteGlun), folate species and total folate accumulating in pods and immature seeds of winged beans from 9 cultivars and 7 growth stages. 5-CH3-H4PteGlun and folate species were determined with a UHPLC-MS/MS method. Accurate determination of 5-CH3-H4PteGlun and folate species was optimized and validated according to EMA guidelines including method selectivity, sensitivity, linearity, accuracy, precision, matrix effect and carry-over. The level of total folate is in the range of 73-200 g/100 g in the pods and 33-61 g/100 g in the immature seeds. The predominant folate species in winged beans is 5-CH3-H4PteGlu1. 5-CH3-H4PteGlu5 is the major polyglutamyl folate derivative. The level of total folate is increased about 4 fold with advancing maturity. For pods, the chain length is increased with growth which shifts from 5-CH3-H4PteGlu1 in the early stage to 5-CH3-H4PteGlu5 and 5-CH3-H4PteGlu6 in the 7th stage. Our findings demonstrate that winged beans are good source of folate. The validated UHPLC-MS/MS method allows for the determination of 5-CH3-H4PteGlun and folate species from other vegetable matrices.

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Folate analysis in complex food matrices: Use of a recombinant Arabidopsis γ-glutamyl hydrolase for folate deglutamylation

Cited by 25 publications

References 32 publications

Metabolic engineering of folate and its precursors in Mexican common bean (Phaseolus vulgaris L.)

Metabolic engineering of folate and its precursors in Mexican common bean (Phaseolus vulgaris L.)

In Vivo Rate of Formaldehyde Condensation with Tetrahydrofolate

Quantification of Total Folate, Folate Species and Polyglutamyl Folate Distribution in Winged Beans (<i>Psophocarus tetragonolobus</i> (L) DC) from Different Cultivars and Growth Stages by Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry

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