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Plant and Cell Physiology Advance Access published online on August 24, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci189
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received May 11, 2005
Accepted August 13, 2005

Regular Paper

Glycoengineering of Cyanobacterial Thylakoid Membranes for Future Studies on the Role of Glycolipids in Photosynthesis

Georg Hölzl 1, Ulrich Zähringer 2, Dirk Warnecke 1*, and Ernst Heinz 1

1 Biozentrum Klein Flottbek, University of Hamburg, Ohnhorststr. 18, 22609 Hamburg, Germany
2 Department of Immunochemistry and Biochemical Microbiology, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Parkalle 1-40, 23845 Borstel, Germany

* To whom correspondence should be addressed.
Dirk Warnecke, E-mail: warnecke{at}botanik.uni-hamburg.de


  Abstract

The lipid composition of thylakoid membranes is conserved from cyanobacteria to angiosperms. The predominating components are monogalactosyl- and digalactosyldiacylglycerol. In cyanobacteria their biosynthesis starts with the formation of monoglucosyldiacylglycerol which is C4-epimerized to the corresponding galactolipid, whereas in plastids monogalactosyldiacylglycerol is formed from the beginning. This points to specific functions of galactolipids in thylakoids. We wanted to investigate the question, whether galactolipids can be replaced by glycosyldiacylglycerols with different headgroups regarding epimeric and anomeric details as well as the attachment point of the terminal hexose in diglycosyldiacylglycerols. For this purpose putative glycosyltransferase sequences were identified in data bases to be used for functional expression in various host organisms. From eighteen newly identified sequences, four turned out to encode glycosyltransferases catalyzing final steps in glycolipid biosynthesis: two {alpha}-glucosyltransferases, one {beta}-galactosyltransferase and one {beta}-glucosyltransferase. Their functional annotation was based on detailed structural characterization of the new glycolipids formed in the transformant hosts as well as on in vitro enzymatic assays. The expression of {alpha}-glucosyltransferases in the cyanobacterium Synechococcus resulted in the accumulation of the new {alpha}-galactosyldiacylglycerol which is ascribed to epimerisation of the corresponding glucolipid. The expression of the {beta}-glucosyltransferase led to a high proportion of new {beta}-glucosyl-(1->6)-{beta}-galactosyldiacylglycerol nearly completely replacing the native digalactosyldiacylglycerol. These results demonstrate that modifications of the glycolipid pattern in thylakoids are possible.

Keywords: Chloroflexus aurantiacus; Deinococcus radiodurans; galactolipid; glycosyltransferase; Synechococcus.
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