Purification, sequencing and characterization of phospholipase D from Indian mustard seeds

Khatoon, Hafeeza; Mansfeld, Johanna; Schierhorn, Angelika; Ulbrich‐Hofmann, Renate

doi:10.1016/j.phytochem.2015.05.022

Cited by 18 publications

(9 citation statements)

References 57 publications

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“…The protein concentration in crude was found to be 14 mg and in purified sample was 0.2 mg. A 53.2 purification fold was obtained with recovery of 75.8%. From 5g of sample about 0.2 mg of purified PLD was obtained with specific activity of 4575 (U mg -1 ) which are not exactly same but near to the previously published on PLD from Streptomyces sanglieri [20] and from Indian mustard seed [18]. SDS-PAGE was done for molecular mass determination.…”

Section: Resultssupporting

confidence: 55%

Isolation, purification and characterization of Phospholipase D from almond (Amygdalus spinosissima) from Ziarat, Balochistan

Baloch¹

2022

PAB

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This study was carried out for the isolation, purification, and characterization of enzyme phospholipase D (PLD) from Amygdalus spinosissima Bunge (wild thorny almond). It was observed that the crude extract showed higher values of enzyme activity 1206 U than that of purified which was found to be 915 U. The optimum time for extraction was 1.5 hours, after which the activity started decreasing. Purification of enzyme was done by hydrophobic interaction chromatography with a specific effect of calcium using Octyl Sepharose. In the absence of calcium ion, the fraction eluted from the column showed high enzymatic activity. The highest specific activity (4575 Umg -1 ), purification fold (53.2) and yield (75.8%) were obtained. Molecular weight was estimated as 62 kDa by SDS-PAGE (sodium dodecyl sulfatepolyacrylamide gel electrophoresis) analysis. Highest activity of PLD was observed at 50℃ and at 6 pH. Calcium ion stability of PLD activity was found at 40-90 mM. It is concluded from the present study that PLD has numerous industrial applications. It can be extracted and purified from A. spinosissima which is a cheap and easily available source at industrial scale.

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Section: Resultssupporting

confidence: 55%

Isolation, purification and characterization of Phospholipase D from almond (Amygdalus spinosissima) from Ziarat, Balochistan

Baloch¹

2022

PAB

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“…Additionally, Nelson reported that PtdEtn conversion yield was only 71−77% of that with PtdCho (Nelson & Cox, ). It was found that an alkyl substitution at the sn‐2 position of PtdCho affects its ability to act as a substrate for cabbage PLD (Bugaut, Kuksis, & Myher, ; Khatoon, Mansfeld, Schierhorn, & Ulbrich‐Hofmann, ). Considering that the PtdSer yield obtained using sea cucumber PlsEtn was significantly lower than that obtained using sea cucumber PtdEtn, these results are consistent with our hypothesis that ether‐phospholipids would hinder the transphosphatidylation of PLD.…”

Section: Discussionmentioning

confidence: 99%

Reaction Specificity of Phospholipase D Prepared from Acinetobacter radioresistens a2 in Transphosphatidylation

Cao

Liu

Tian

et al. 2018

Lipids

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Phospholipase D (PLD) can react with phospholipids as substrates, generally phosphatidylcholine (PtdCho), and the PLD-substrate intermediate can be cleaved by another alcohol, resulting in transphosphatidylation of the substrate, which can be used in the production of special lipids. In this study, the reaction conditions affecting the transphosphatidylation of PtdCho with serine were optimized and the reaction specificity of a novel PLD prepared from Acinetobacter radioresistens a2 was evaluated for transphosphatidylation with a variety of phospholipid substrates and head group donors. Based on the yield of phosphatidylserine, experimental kinetic data, maximum transphosphatidylation rate, and kinetic constant, the specificity of PLD in transphosphatidylation was found to be affected by unsaturated fatty-acid phospholipid substrates. The catalytic efficiency of PLD prepared from A. radioresistens a2 on the synthesis of natural phospholipids is on the order of l-serine > ethanolamine and glycerol ≫ inositol. Moreover, it was found that the transphosphatidylation of PtdCho with saccharides was related to the length of the carbon chain and the number of saccharide units.

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“…[6][7][8][9] PLD (EC 3.1.4.4) is an enzyme that hydrolyzes phospholipids (PLs) into phosphatidic acids (PA) and free alcohol. 10 PLD is naturally distributed in mammals, 11 plants, 12 yeast, 13 and bacteria. [14][15][16] In addition to the important regulatory functions in organisms related to cytomembrane construction, vesicle trafficking, and signal transduction, PLD plays an indispensable role in synthesizing various scarce PLs, 10,17,18 such as PS, phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidyl-tyrosol (PT), and phosphatidylinositol (PI), which are relatively difficult to be obtained in pure forms and on large scale by chemical purification from raw materials.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a phospholipase D from Streptomyces cinnamoneum SK43.003 suitable for phosphatidylserine synthesis

Zhou

Duan

et al. 2021

Biotech and App Biochem

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A phospholipase D high producing strain with transphosphatidylation activity that is suitable for phosphatidylserine synthesis was screened by our laboratory and named as Streptomyces cinnamoneum SK43.003. The enzyme structural and biochemical properties were investigated using the molecular biology method. A 1521‐bp fragment of the phospholipase D gene from Streptomyces cinnamoneum SK43.003 was amplified by PCR and encoded for 506 amino acids. The primary structure contained two conserved HKD and GG/S motifs. The pld gene was cloned and expressed in Escherichia coli. The purified enzyme exhibited the highest activity at a pH value of 6.0 andtemperature of 60°C. The enzyme was stable within a pH range of 4–7 for 24 h or at temperatures below 50°C. In addition, Triton X‐100, Fe2+, and Al3+ were beneficial to the enzyme activity, whereas Zn2+ and Cu2+ dramatically inhibited its activity. In a two‐phase system, the enzyme could convert phosphatidylcholine to phosphatidylserine with a 92% transformation rate.

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Purification, sequencing and characterization of phospholipase D from Indian mustard seeds

Cited by 18 publications

References 57 publications

Isolation, purification and characterization of Phospholipase D from almond (Amygdalus spinosissima) from Ziarat, Balochistan

Isolation, purification and characterization of Phospholipase D from almond (Amygdalus spinosissima) from Ziarat, Balochistan

Reaction Specificity of Phospholipase D Prepared from Acinetobacter radioresistens a2 in Transphosphatidylation

Characterization of a phospholipase D from Streptomyces cinnamoneum SK43.003 suitable for phosphatidylserine synthesis

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