Mining of a phospholipase D and its application in enzymatic preparation of phosphatidylserine

Bioengineered
Wen-Bin ZhouJin-Song Shi

Abstract

Phosphatidylserine (PS) is useful as the additive in industries for memory improvement, mood enhancement and drug delivery. Conventionally, PS was extracted from soybeans, vegetable oils, egg yolk, and biomass; however, their low availability and high extraction cost were limiting factors. Phospholipase D (PLD) is a promising tool for enzymatic synthesis of PS due to its transphosphatidylation activity. In this contribution, a new and uncharacterized PLD was first obtained from GenBank database via genome mining strategy. The open reading frame consisted of 1614 bp and potentially encoded a protein of 538-amino-acid with a theoretical molecular mass of 60 kDa. The gene was successfully cloned and expressed in Escherichia coli. Its enzymatic properties were experimentally characterized. The temperature and pH optima of PLD were determined to be 60°C and 7.5, respectively. Its hydrolytic activity was improved by addition of Ca2+ at 5 mM as compared with the control. The enzyme displayed suitable transphosphatidylation activity and PS could be synthesized with L-serine and soybean lecithin as substrates under the catalysis of PLD. This PLD enzyme might be a potential candidate for industrial applications in PS production. To the b...Continue Reading

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Citations

Apr 17, 2019·Bioprocess and Biosystems Engineering·Hai-Juan HouJin-Song Shi
Feb 8, 2019·Applied Microbiology and Biotechnology·Sebastián CerminatiMaría Eugenia Castelli
Jan 5, 2020·Applied Microbiology and Biotechnology·Israel García-CanoRafael Jiménez-Flores
Nov 5, 2020·Biomolecules·Vicneswarry DorairajNor Azian Abdul Murad
May 15, 2021·Critical Reviews in Biotechnology·Peng ZhangJin-Song Shi

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Datasets Mentioned

BETA
BAA75216.1

Methods Mentioned

BETA
PCR

Software Mentioned

BLASTP
GraphPad Prism
Protparam
MEGA
Clustal X

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