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Streptomyces Phospholipase D Cloning and Production

  • Yozo NakazawaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 861)

Abstract

The transphosphatidylation catalytic ability of phospholipase D (PLD, EC 3.1.4.4) is a powerful biochemical tool for the acquisition of rare phospholipids (PLs), e.g., phosphatidylglycerol (PG) and phosphatidylserine (PS), and artificial phospholipids, which do not occur in nature. Specifically, actinomycete PLD recognizes not only the alcohols (i.e., glycerol or serine) corresponding to the polar head groups of natural PLs, but also secondary alcohols, aromatic alcohols, saccharides, N-heterocyclic alcohols, and vitamins as acceptors. Therefore, actinomycete PLD is a valuable enzyme in food, cosmetics, fine chemical and pharmaceutical industries. Here, we describe a protocol for the screening for PLD-producing microorganisms, several PLD assays and methods of PLD production–purification and the strategy of cloning of the Streptomyces PLD gene.

Key words

Streptomyces Phospholipase D Screening Purification Cloning Transphos­phatidylation 

Notes

Acknowledgments

The author thanks Dr. Yoshimasa Sagane (Sars International Centre for Marine Molecular Biology, Bergen, Norway) for his excellent advice and encouragement with regard to this work.

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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  1. 1.Department of Food and Cosmetic Science, Faculty of BioindustryTokyo University of AgricultureAbashiriJapan

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