Direct Enzymatic Synthesis of 1-Phosphatidyl-β-D-glucose by Engineered Phospholipase D

2016 ◽  
Vol 1 (13) ◽  
pp. 4121-4125 ◽  
Author(s):  
Arisa Inoue ◽  
Masaatsu Adachi ◽  
Jasmina Damnjanović ◽  
Hideo Nakano ◽  
Yugo Iwasaki
Keyword(s):  
Phospholipase D ◽  
Enzymatic Synthesis

Acyl chain that matters: introducing sn-2 acyl chain preference to a phospholipase D by protein engineering

2019 ◽  
Vol 32 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Jasmina Damnjanović ◽  
Hideo Nakano ◽  
Yugo Iwasaki
Keyword(s):  
Protein Engineering ◽  
Phospholipase D ◽  
Enzymatic Synthesis ◽  
Acyl Chain ◽  
Saturation Mutagenesis ◽  
Structure Model ◽  
Streptomyces Antibioticus ◽  
In The Wild ◽  
Structured Phospholipids ◽  
Experimental Findings

AbstractPhospholipase D (PLD) is an enzyme widely used for enzymatic synthesis of structured phospholipids (PLs) with modified head groups. These PLs are mainly used as food supplements and liposome ingredients. Still, there is a need for an enzyme that discriminates between PLs and lysoPLs, for specific detection of lysoPLs in various specimens and enzymatic synthesis of certain PLs from a mixed substrate. To meet this demand, we aimed at altering sn-2 acyl chain recognition of a PLD, leading to a variant enzyme preferably reacting on lysoPLs, by protein engineering. Based on the crystal structure of Streptomyces antibioticus PLD, W166 was targeted for saturation mutagenesis due to its strong interaction with the sn-2 acyl chain of the PL. Screening result pointed at W166R and W166K PLDs to selectively react on lysophosphatidylcholine (lysoPC), while not on PC. These variants showed a negative correlation between activity and sn-2 chain length of PL substrates. This behavior was not observed in the wild-type (WT)-PLD. Kinetic analysis revealed that the W166R and W166K variants have 7–10 times higher preference to lysoPC compared to the WT-PLD. Additionally, W166R PLD showed detectable activity toward glycero-3-phosphocholine, unlike the WT-PLD. Applicability of the lysoPC-preferring PLD was demonstrated by detection of lysoPC in the mixed PC/lysoPC sample and by the synthesis of cyclic phosphatidic acid. Structure model analyses supported the experimental findings and provided a basis for the structure model-based hypothesis on the observed behavior of the enzymes.

Facile Enzymatic Synthesis of Phosphatidylthreonine Using an Engineered Phospholipase D

2018 ◽  
Vol 120 (6) ◽  
pp. 1800089 ◽  
Author(s):  
Jasmina Damnjanović ◽  
Nozomi Matsunaga ◽  
Masaatsu Adachi ◽  
Hideo Nakano ◽  
Yugo Iwasaki
Keyword(s):  
Phospholipase D ◽  
Enzymatic Synthesis

Directing positional specificity in enzymatic synthesis of bioactive 1-phosphatidylinositol by protein engineering of a phospholipase D

2015 ◽  
Vol 113 (1) ◽  
pp. 62-71 ◽  
Author(s):  
Jasmina Damnjanović ◽  
Chisato Kuroiwa ◽  
Hidetoshi Tanaka ◽  
Ken Ishida ◽  
Hideo Nakano ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Phospholipase D ◽  
Enzymatic Synthesis ◽  
Positional Specificity

Enzymatic synthesis of structural analogs of PAF-acether by phospholipase D-catalysed transphosphatidylation

1992 ◽  
Vol 1123 (3) ◽  
pp. 347-350 ◽  
Author(s):  
Valérie Testet-Lamant ◽  
Brigitte Archaimbault ◽  
Jacqueline Durand ◽  
Michel Rigaud
Keyword(s):  
Phospholipase D ◽  
Enzymatic Synthesis

Enzymatic Synthesis of Enantiomerically Pure Chiral Synthons: Lipase-Catalyzed Resolution of (R/S, 4E)-2-Methyl-4-hexen-l-ol

Synlett ◽  
1991 ◽  
Vol 1991 (04) ◽  
pp. 310-312
Author(s):  
Patrizia Ferraboschi ◽  
Daria Brembilla ◽  
Paride Grisenti ◽  
Enzo Santaniello
Keyword(s):  
Enzymatic Synthesis ◽  
Enantiomerically Pure

A common precursor to both prebiotic and biological pathways to RNA nucleotides

2018 ◽  
Author(s):  
Andrea Pérez-Villa ◽  
Thomas Georgelin ◽  
Jean-François Lambert ◽  
Marie-Christine Maurel ◽  
François Guyot ◽  
...  
Keyword(s):  
Enzymatic Synthesis ◽  
De Novo ◽  
Life Forms ◽  
Hydrothermal Conditions ◽  
Modern Life ◽  
Ab Initio Simulations ◽  
Spontaneous Formation ◽  
Common Precursor ◽  
Open Issue ◽  
Anomeric Carbon

Understanding the mechanism of spontaneous formation of ribonucleotides under realistic prebiotic conditions is a key open issue of origins-of-life research. In cells, <i>de novo</i> and salvage nucleotide enzymatic synthesis combines 5-phospho-α -D-ribose-1-diphosphate ( α-PRPP) and nucleobases. Interestingly, these reactants are also known as prebiotically plausible compounds. Combining ab initio simulations with mass spectrometry experiments, we compellingly demonstrate that nucleobases and α -PRPP spontaneously combine, through the same facile mechanism, forming both purine and pyrimidine ribonucleotides, under mild hydrothermal conditions. Surprisingly, this mechanism is very similar to the biological one, and yields ribonucleotides with the same anomeric carbon chirality as in biological systems. These results suggest that natural selection might have optimized – through enzymes – a pre-existing ribonucleotide formation mechanism, carrying it forward to modern life forms.

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