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

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.

scholarly journals Synthesis of Human Milk Oligosaccharides: Protein Engineering Strategies for Improved Enzymatic Transglycosylation

Molecules ◽  
2019 ◽  
Vol 24 (11) ◽  
pp. 2033 ◽  
Author(s):  
Birgitte Zeuner ◽  
David Teze ◽  
Jan Muschiol ◽  
Anne S. Meyer
Keyword(s):  
Protein Engineering ◽  
Human Milk ◽  
Active Site ◽  
Enzymatic Synthesis ◽  
Spatial Arrangement ◽  
Glycoside Hydrolases ◽  
High Yield ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Enzyme Active Site

Human milk oligosaccharides (HMOs) signify a unique group of oligosaccharides in breast milk, which is of major importance for infant health and development. The functional benefits of HMOs create an enormous impetus for biosynthetic production of HMOs for use as additives in infant formula and other products. HMO molecules can be synthesized chemically, via fermentation, and by enzymatic synthesis. This treatise discusses these different techniques, with particular focus on harnessing enzymes for controlled enzymatic synthesis of HMO molecules. In order to foster precise and high-yield enzymatic synthesis, several novel protein engineering approaches have been reported, mainly concerning changing glycoside hydrolases to catalyze relevant transglycosylations. The protein engineering strategies for these enzymes range from rationally modifying specific catalytic residues, over targeted subsite −1 mutations, to unique and novel transplantations of designed peptide sequences near the active site, so-called loop engineering. These strategies have proven useful to foster enhanced transglycosylation to promote different types of HMO synthesis reactions. The rationale of subsite −1 modification, acceptor binding site matching, and loop engineering, including changes that may alter the spatial arrangement of water in the enzyme active site region, may prove useful for novel enzyme-catalyzed carbohydrate design in general.

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

Phospholipase D from Streptoverticillium cinnamoneum: protein engineering and application for phospholipid production

2003 ◽  
Vol 23 (2-6) ◽  
pp. 107-115 ◽  
Author(s):  
Chiaki Ogino ◽  
Shun’ichi Kuroda ◽  
Shinji Tokuyama ◽  
Akihiko Kondo ◽  
Nobuaki Shimizu ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Phospholipase D

Protein engineering of thioether monooxygenase to improve its thermostability for enzymatic synthesis of chiral sulfoxide

2021 ◽  
Vol 509 ◽  
pp. 111625
Author(s):  
Peng Zhao ◽  
Shi-Miao Ren ◽  
Feng Liu ◽  
Yu-Cong Zheng ◽  
Na Xu ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Enzymatic Synthesis

Isolation of Phospholipase D Mutants Having Phosphatidylinositol-Synthesizing Activity with Positional Specificity onmyo-Inositol

ChemBioChem ◽  
2009 ◽  
Vol 10 (3) ◽  
pp. 559-564 ◽  
Author(s):  
Atsushi Masayama ◽  
Kaori Tsukada ◽  
Chika Ikeda ◽  
Hideo Nakano ◽  
Yugo Iwasaki
Keyword(s):  
Phospholipase D ◽  
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

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
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