Simple and Rapid Method for the Selective Detection of Individual Lipoxygenase Isoenzymes in Soybean Seeds

Suda, Ikuo; Hajika, Makita; Nishiba, Yoichi; Furuta, Shu; Igita, Kazunori

doi:10.1021/jf00051a034

Cited by 61 publications

(49 citation statements)

References 13 publications

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“…The lipoxygenase assay was performed based on the decolorization of methyleneblue during the enzymatic reaction (Anthon & Barrett, 2003;Suda et al, 1995). Linoleic acid solution as a 50 mM stock was first prepared in a volume of 1.0 mL, containing linoleic acid (56 mg), Tween 20 (112 mg), 1 N NaOH (24 mL), and water (811.2 mL), and stored at À20 C under N 2 gas.…”

Section: Lipoxygenase Inhibitionmentioning

confidence: 99%

Inhibitory effects of the ethyl acetate extract from bulbs ofScilla scilloideson lipoxygenase and hyaluronidase activities

Nishida

Sugahara

Wada

et al. 2014

Pharmaceutical Biology

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Context: Scilla scilloides Druce (Liliaceae) is a folk medicine to treat dermal inflammation; however, the medicinal properties of this plant have not been completely established. Objective: The current study investigates the potent anti-inflammatory effects of S. scilloides bulbs for its traditional usage using lipoxygenase and hyaluronidase as the inflammation model. To gain insight into the active constituents, nine homoisoflavones (1-9) were subsequently tested. Materials and methods: Lipoxygenase and hyaluronidase inhibition of ethyl acetate extract from the bulbs of this plant within 2000 mg/mL or homoisoflavones within 1000 mM were determined by colorimetric methods. RAW264.7 cells were incubated with 10 or 50 mM homoisoflavones plus lipopolysaccharide (LPS) for 24 h. The culture media were collected and analyzed for determination of the nitric oxide (NO) level by the colorimetric Griess method to measure the extent of inflammation. Results: The extract exhibited inhibitory effects on lipoxygenase and hyaluronidase activities with IC 50 values 31.5 and 169 mg/mL, respectively. Among the nine homoisoflavones tested, four (1 and 3-5) resulted in 79.3-97.9% higher lipoxygenase inhibition than 6.7-32.7% of the others at 500 mM. Calculated IC 50 values indicated 5 as the compound responsible for strong lipoxygenase inhibition with 15.8 mM as the IC 50 value. In the hyaluronidase assay, all homoisoflavones tested at 1000 mM demonstrated 16.2-58.0% inhibition. Incubating the cells in the presence of all nine homoisoflavones tested at 50 mM significantly suppressed the NO production, downward to 1.5-66.0%, in the LPS-activated macrophage cells as a model. Discussion and conclusion: These results may indicate a potential role of S. scilloides for anti-inflammatory purposes.

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Section: Lipoxygenase Inhibitionmentioning

confidence: 99%

Inhibitory effects of the ethyl acetate extract from bulbs ofScilla scilloideson lipoxygenase and hyaluronidase activities

Nishida

Sugahara

Wada

et al. 2014

Pharmaceutical Biology

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“…The fatty acid composition was determined by gas liquid chromatography of fatty acid methyl esters according to the modified method of Bannon et al (1982) Estimates of ω-6 and ω-3 desaturase activities (%) were derived from measurements of unsaturated fatty acids contents (Cherif et al 1975) Suda et al (1995). The 3lx lines were confirmed by SDS polyacrylamide gel electrophoresis (Kitamura et al 1983).…”

Section: Analysis Of Seed LX and La Contentmentioning

confidence: 99%

Relationships and inheritance of linolenic acid and seed lipoxygenases in soybean crosses designed to combine these traits

Reinprecht¹,

Rajcan²,

Poysa³

et al. 2005

Can. J. Plant Sci.

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Reinprecht, Y., Rajcan, I., Poysa, V. W., Ablett, G. R. and Pauls, K. P. 2005. Relationships and inheritance of linolenic acid and seed lipoxygenases in soybean crosses designed to combine these traits. Can. J. Plant Sci. 85: 593-602. To improve oxidative stability of soybean oil and reduce off-flavours, we previously developed low linolenic acid, lipoxygenase-free (LLA.3lx) soybean germplasm. The objectives of this study were to characterize the patterns of inheritance and determine the relationships between the low linolenic acid (LLA) trait derived from the lines RG10 and PI 361088B and seed lipoxygenase nulls (3lx) from a triple null line OX948 that were used to create the new LLA.3lx germplasm. Reciprocal crosses between RG10 and OX948 and between PI 361088B and OX948 were made and populations derived from them were evaluated for their fatty acid profiles and seed lipoxygenases (LX) at the F 2 , F 5 and F 6 generations. Both RG10 and PI 361088B contain a single gene that controls linolenic acid (LA) content with alleles acting in an additive manner. No significant cytoplasmic effects were observed on LA content. The LLA trait was highly heritable in both RG10 × OX948 (RO) and PI 361088B × OX948 (PO) crosses and stable in different environments (Belize and Ontario, Canada). The three seed LX were each controlled by single genes, but lipoxygenase 1 (Lx1) and lipoxygenase 2 (Lx2) were tightly linked. No significant correlations were observed between LA content and seed LX. The simple and independent inheritance of LA and seed LX in these crosses should simplify breeding for soybean cultivars that are resistant to oxidative degradation. La teneur en LA ne subit aucune influence significative du cytoplasme. Le caractère LLA se transmet très facilement dans les croisements RG10 × OX948 (RO) et PI 361088B × OX948 (PO), et reste stable dans divers milieux (Bélize, Ontario au Canada). Les trois lipoxygénases de la graine sont régulées par des gènes uniques, mais la lipoxygénase 1 (Lx1) et la lipoxygénase 2 (Lx2) sont étroitement associées. Les auteurs n'ont observé aucune corrélation significative entre la teneur en LA et les LX de la graine. L'héritabilité simple et indépendante des deux caractères dans les croisements devrait simplifier l'hybridation de cultivars résistants à la dégradation par l'oxydation.

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“…As sementes da geração F2 foram colhidas e identificadas para presença de lipoxigenases por meio de testes colorimétricos (Suda et al, 1995) e determinação da atividade de lipoxigenases. Foi realizada, também, a determinação da composição de ácidos graxos da fração óleo por cromatografia gasosa, com a utilização de metodologia de Bubeck et.…”

Section: Methodsunclassified

Lipoxigenases e teor de ácido linolênico relacionados à qualidade de sementes de soja

Oliveira

Piovesan

José

et al. 2006

Rev. bras. sementes

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Os teores de ácidos graxos da fração óleo das sementes de soja não têm sido considerados na avaliação da qualidade das sementes em trabalhos de pesquisa recentes. O objetivo deste trabalho foi determinar o efeito da ausência de lipoxigenases e baixo teor de ácido linolênico na qualidade de sementes de soja colhidas em diferentes épocas após o estádio R8. Quatro "bulks" de sementes de soja foram formados, com as características: (a) presença de lipoxigenases e teor normal de ácido linolênico, (b) presença de lipoxigenases e baixo teor de ácido linolênico, (c) ausência de lipoxigenases e teor normal de ácido linolênico e (d) ausência de lipoxigenases e baixo teor de ácido linolênico. Sementes de cada "bulk" da geração F5 foram colhidas nos estádios R8, R8+10, R8+20 e R8+30 dias. A qualidade fisiológica das sementes foi avaliada através dos testes de germinação, envelhecimento acelerado e índice de velocidade de emergência. Foi determinado também os teores de ácido linolênico das sementes. Sementes dos "bulks" com baixo teor de ácido linolênico apresentaram maior germinação e menor número de plântulas anormais após o estádio R8+20 dias. Sementes dos "bulks" com lipoxigenases apresentaram maior velocidade de emergência. O baixo teor de ácido linolênico presente na fração óleo da semente de soja favorece a produção de sementes de melhor qualidade e a presença de lipoxigenase favorece a maior velocidade de emergência de plântulas.

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Simple and Rapid Method for the Selective Detection of Individual Lipoxygenase Isoenzymes in Soybean Seeds

Cited by 61 publications

References 13 publications

Inhibitory effects of the ethyl acetate extract from bulbs ofScilla scilloideson lipoxygenase and hyaluronidase activities

Inhibitory effects of the ethyl acetate extract from bulbs ofScilla scilloideson lipoxygenase and hyaluronidase activities

Relationships and inheritance of linolenic acid and seed lipoxygenases in soybean crosses designed to combine these traits

Lipoxigenases e teor de ácido linolênico relacionados à qualidade de sementes de soja

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