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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Yang SF, Freer S, Benson AA (1967) Transphosphatidylation by phospholipase D. J Biol Chem 242:477–484
Morris AJ, Frohman MA, Engebrecht J (1997) Measurement of phospholipase D activity. Anal Biochem 252:1–9
D’Arrigo P, de Ferra L, Piergianni V et al (1996) Preparative transformation of natural phospholipids catalyzed by phospholipase D from Streptomyces. J Chem Soc Perkin Trans I 21:2651–2656
D’Arrigo P, de Ferra L, Piergianni V et al (1994) Phospholipase D from Streptomyces catalyses the transfer of secondary alcohols. J Chem Soc Chem Commun 14:1709–1710
Takami M, Hidaka N, Suzuki Y (1994) Phospholipase D-catalyzed synthesis of phosphatidyl aromatic compounds. Biosci Biotechnol Biochem 58:2140–2144
Kokusyo Y, Tsunoda A, Kato S et al (1993) Production of various phosphatidylsaccharides by phospholipase D from Actinomadura sp. strain No. 362. Biosci Biotechnol Biochem 57:1302–1305
Aurich I, Dürrschmidt P, Hirche F et al (1997) Transesterification of alkylphosphate esters by phospholipase D. Biotechnol Lett 19:875–879
Nagao A, Ishida N, Terao J (1991) Synthesis of 6-phosphatidyl-L-ascorbic acid by phospholipase D. Lipids 26:390–394
Koga T, Nagao A, Terao J et al (1994) Synthesis of a phosphatidyl derivative of vitamin E and its antioxidant activity in phospholipid bilayers. Lipids 29:83–89
Wang P, Schuster M, Wang YF et al (1993) Synthesis of phospholipid inhibitor conjugates by enzymatic transphosphatidylation with phospholipase D. J Am Chem Soc 115:10487–10491
Shuto S, Itoh H, Sakai A et al (1995) Nucleosides and nucleotides - CXXXVII. Antitumor phospholipids with 5-fluorouridine as a cytotoxic polarhead: Synthesis of 5’-phosphatidyl-5-fluorouridines by phospholipase D-catalyzed transphosphatidylation. Bioorg Med Chem 3:235–243
Nakazawa Y, Uchino M, Sagane Y et al (2009) Isolation and characterization of actinomycetes strains that produce phospholipase D having high transphosphatidylation activity. Microbiol Res 164:43–48
Nakazawa Y, Suzuki R, Uchino M et al (2010) Identification of actinomycetes producing phospholipase d with high transphosphatidylation activity. Curr Microbiol 60:365–372
Nakazawa Y, Sagane Y, Kikuchi T et al (2010) Purification, biochemical characterization, and cloning of phospholipase D from Streptomyces racemochromogenes strain 10–3. Protein J 29:598–608
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media New York
About this protocol
Cite this protocol
Nakazawa, Y. (2012). Streptomyces Phospholipase D Cloning and Production. In: Sandoval, G. (eds) Lipases and Phospholipases. Methods in Molecular Biology, vol 861. Humana Press. https://doi.org/10.1007/978-1-61779-600-5_12
Download citation
DOI: https://doi.org/10.1007/978-1-61779-600-5_12
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-599-2
Online ISBN: 978-1-61779-600-5
eBook Packages: Springer Protocols