A - Papers appearing in refereed journals
Sperling, P., Libisch, B., Zahringer, U., Napier, J. A. and Heinz, E. 2001. Functional identification of a delta8-sphingolipid desaturase from Borago officinalis. Archives of Biochemistry and Biophysics. 388 (2), pp. 293-298. https://doi.org/10.1006/abbi.2001.2308
Authors | Sperling, P., Libisch, B., Zahringer, U., Napier, J. A. and Heinz, E. |
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Abstract | The similarities between Δ12- and Δ15-fatty acyl desaturase sequences were used to construct degenerate primers for PCR experiments with cDNA transcribed from mRNA of developing borage seeds. Screening of a borage seed cDNA library with an amplified DNA fragment resulted in the isolation of a full-length cDNA corresponding to a deduced open-reading frame of 446 amino acids. The protein showed high similarity to plant Δ8-sphingolipid desaturases as well as to the Δ6-fatty acyl desaturase from Borago officinalis. The sequence is characterized by the presence of a N-terminal cytochrome b5 domain. Expression of this open-reading frame in Saccharomyces cerevisiae resulted in the formation of Δ8-trans/cis-phytosphingenines not present in wild-type cells, as shown by HPLC analysis of sphingoid bases as their dinitrophenyl derivatives. GLC-MS analysis of the methylated di-0-trimethylsilyl ether derivatives confirmed the presence of Δ8-stereoisomers of C18- and C20-phytosphingenine. Furthermore, Northern blotting showed that the gene encoding a stereo-unselective Δ8-sphingolipid desaturase is primarily expressed in young borage leaves. |
Keywords | cytochrome b5; desaturase; Borago officinalis; GLC-MS; phytosphingenine; reversed-phase HPLC; Saccharomyces cerevisiae; sphingolipids |
Year of Publication | 2001 |
Journal | Archives of Biochemistry and Biophysics |
Journal citation | 388 (2), pp. 293-298 |
Digital Object Identifier (DOI) | https://doi.org/10.1006/abbi.2001.2308 |
Open access | Published as non-open access |
Funder project or code | 415 |
503 | |
Project: 3258 | |
Output status | Published |
Publication dates | |
Online | 25 May 2002 |
Copyright license | Publisher copyright |
ISSN | 0003-9861 |
Publisher | Elsevier Science Inc |
Permalink - https://repository.rothamsted.ac.uk/item/8885w/functional-identification-of-a-delta8-sphingolipid-desaturase-from-borago-officinalis
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