The purification and characterization of fatty acid hydroperoxide lyase in sunflower

Itoh, Aya; Vick, Brian

doi:10.1016/s0005-2760(98)00161-1

Cited by 25 publications

(37 citation statements)

References 40 publications

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“…Monsan and Combes [10] have suggested that the protective effect of several chemical additives on the enzyme thermal stability is attributed to their effect on the water activity; indeed, the high interaction of additives with water molecules will result in the formation of water clusters and then in the reduction of the amount of free water [31,32]. The thermal stability profiles of HPL are in agreement with those reported for HPL activity from other sources [16,31,33].…”

Section: Effect Of Additives On Hpl Thermal Stabilitysupporting

confidence: 83%

“…Compared to glycine and dithiothreitol, the effect of Triton X-100 was independent on its concentration; a specific activity of 0.14 µmol converted HPOD/(min mg protein) was obtained at a wide range of Triton X-100 concentrations, varying from 0.25% to 1.5% (w/v). Detergents are generally added to the HPL enzymatic extract at percentage ranging from 0.1% to 0.5% (w/v) [11,13]; they are regarded as inactivators for HPL activity at a concentration greater than 1% (w/v) [31].…”

Section: Effect Of Selected Additives On Hpl Activitymentioning

confidence: 99%

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Activation and Stabilization of The Hydroperoxide Lyase Enzymatic Extract from Mint Leaves (Mentha spicata) Using Selected Chemical Additives

Akacha

Karboune

Gargouri

et al. 2009

Appl Biochem Biotechnol

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The effects of selected lyoprotecting excipients and chemical additives on the specific activity and the thermal stability of the hydroperoxide lyase (HPL) enzymatic extract from mint leaves were investigated. The addition of KCl (5%, w/w) and dextran (2.5%, w/w) to the enzymatic extract, prior to lyophilization, increased the HPL specific activity by 2.0- and 1.2-fold, respectively, compared to the control lyophilized extract. From half-life time (t (1/2)), it can be seen that KCl has enhanced the HPL stability by 1.3- to 2.3-fold, during long-period storage at -20 degrees Celsius and 4 degrees Celsius. Among the selected additives used throughout this study, glycine appeared to be the most effective one. In addition to the activation effect conferred by glycine, it also enhanced the HPL thermal stability. In contrast, polyhydroxyl-containing additives were not effective for stabilizing the HPL enzymatic extract. On the other hand, there was no signification increase in HPL activity and its thermal stability with the presence of Triton X-100. The results also showed that in the presence of glycine (10%), the catalytic efficiency of HPL was increased by 2.45-fold than that without additive.

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Section: Effect Of Additives On Hpl Thermal Stabilitysupporting

confidence: 83%

Section: Effect Of Selected Additives On Hpl Activitymentioning

confidence: 99%

Activation and Stabilization of The Hydroperoxide Lyase Enzymatic Extract from Mint Leaves (Mentha spicata) Using Selected Chemical Additives

Akacha

Karboune

Gargouri

et al. 2009

Appl Biochem Biotechnol

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“…For the 13-HPL from green bell pepper fruits, Shibata et al (12) showed also that 13-HPL could be resolved into two isoforms by hydroxylapatite chromatography; in this case, both isoforms had molecular masses of 55 kD (12). Similar to the lyases of sunflower (13), the apparent molecular weight of the guava 13-HPL as determined by gel filtration is in the order of 200,000 kD, implying a tetrameric structure for the native enzyme. This is in agreement with Olias et al (10) who reported that 13-HPL from soybean seedlings had a native molecular mass of 240-260 kD and a subunit size of 62 kD.…”

Section: Discussionmentioning

confidence: 85%

“…It is also possible that other isoforms of hydroperoxide lyase might exist in guava. Of the two hydroperoxide lyases purified by Itoh and Vick (13) from sunflower hypocotyl, one resembled the guava and pepper enzymes and a second showed equal reactivity with the C18:2 analog, 13-hydroperoxylinoleic acid.…”

Section: Discussionmentioning

confidence: 99%

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Purification, molecular cloning, and expression of the gene encoding fatty acid 13‐hydroperoxide lyase from guava fruit (Psidium guajava)

Tijet

Wäspi

Gaskin³

et al. 2000

Lipids

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Guava fruit was identified as a particularly rich source of 13-hydroperoxide lyase activity. The enzyme proved stable to chromatographic procedures and was purified to homogeneity. Based on gel filtration and gel electrophoresis, the native enzyme appears to be a homotetramer with subunits of 55 kD. Starting with primers based on the peptide sequence, the enzyme was cloned by polymerase chain reaction with 3' and 5' rapid amplification of cDNA ends. The sequence shows approximately 60-70% identity to known 13-hydroperoxide lyases and is classified in cytochrome P450 74B subfamily as CYP74B5. The cDNA was expressed in Escherichia coli (BL21 cells), with optimal enzyme activity obtained in the absence of isopropyl-beta-D-thiogalactopyranoside and delta-aminolevulinic acid. The expressed enzyme metabolized 13(S)-hydroperoxylinolenic acid over 10-fold faster than 13(S)-hydroperoxylinoleic acid and the 9-hydroperoxides of linoleic and linolenic acids. 13(S)-Hydroperoxylinolenic acid was converted to 12-oxododec-9(Z)-enoic acid and 3(Z)-hexenal, as identified by gas chromatography-mass spectrometry. The turnover number with this substrate, with enzyme concentration estimated from the Soret absorbance, was approximately 2000/s, comparable to values reported for the related allene oxide synthases. Distinctive features of the guava 13-hydroperoxide lyase and related cytochrome P450 are discussed.

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Production of tomato flavor volatiles from a crude enzyme preparation using a hollow-fiber reactor

Cass

Schade

Robinson

et al. 2000

Biotechnol. Bioeng.

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In recent years there has been an increase in the interest in the production of compounds by isolation from natural sources or through processes that can be deemed “natural”. This is of particular interest in the food and beverage industry for flavors and aromas. Hexanal, organoleptically known to possess “green character”, is of considerable commercial interest. The objective of this study was to determine if the enzyme template known to be responsible for the synthesis of hexanal from linoleic acid (18:2) in tomato fruits could be harnessed using a hollow‐fiber reactor. A hollow‐fiber reactor system was set up and consisted of a XAMPLER ultrafiltration module coupled to a reservoir. The enzyme template was extracted from ripe tomato fruits and processed through an ultrafiltration unit (NMWC of 100 kDa) to produce a retentate enriched in soluble and membrane‐associated lipoxygenase (LOX) and hydroperoxide lyase (HPL). This extract was recirculated through the lumen of the hollow‐fiber ultrafiltration unit with the addition of substrate in the form of linoleic acid, with buffer addition to the reaction flask to maintain a constant retentate volume. Product formation was measured in the permeate using solid phase microextraction (SPME) developed for this system. At exogenous substrate concentrations of 16 mM and a transmembrane pressure of 70 kPa, hexanal production rates are in the order of 5.1 μg/min. Addition of Triton X‐100 resulted in membrane fouling and reduced flux. The reactor system has been run for periods of up to 1 week and has been shown to be stable over this period. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 67: 372–377, 2000.

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The purification and characterization of fatty acid hydroperoxide lyase in sunflower

Cited by 25 publications

References 40 publications

Activation and Stabilization of The Hydroperoxide Lyase Enzymatic Extract from Mint Leaves (Mentha spicata) Using Selected Chemical Additives

Activation and Stabilization of The Hydroperoxide Lyase Enzymatic Extract from Mint Leaves (Mentha spicata) Using Selected Chemical Additives

Purification, molecular cloning, and expression of the gene encoding fatty acid 13‐hydroperoxide lyase from guava fruit (Psidium guajava)

Production of tomato flavor volatiles from a crude enzyme preparation using a hollow-fiber reactor

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