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PMID:22385412

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Citation

Kikuta, Y, Ueda, H, Takahashi, M, Mitsumori, T, Yamada, G, Sakamori, K, Takeda, K, Furutani, S, Nakayama, K, Katsuda, Y, Hatanaka, A and Matsuda, K (2012) Identification and characterization of a GDSL lipase-like protein that catalyzes the ester-forming reaction for pyrethrin biosynthesis in Tanacetum cinerariifolium- a new target for plant protection. Plant J. 71:183-93

Abstract

Although natural insecticides pyrethrins produced by Tanacetum cinerariifolium are used worldwide to control insect pest species, little information is known of their biosynthesis. From the buds of T. cinerariifolium, we have purified a protein that is able to transfer the chrysanthemoyl group from the coenzyme A (CoA) thioester to pyrethrolone to produce pyrethrin I and have isolated cDNAs that encode the enzyme. To our surprise, the active principle was not a member of a known acyltransferase family but a member of the GDSL lipase family. The recombinant enzyme (TcGLIP) was expressed in Escherichia coli and displayed the acyltransferase reaction with high substrate specificity, recognized the absolute configurations of three asymmetric carbons and also showed esterase activity. A S40A mutation in the Block I domain reduced both acyltransferase and esterase activities, which suggested an important role of this serine residue in these two activities. The signal peptide directed the localization of TcGLIP::enhanced green fluorescent protein (EGFP) fusion, as well as EGFP, to the extracellular space. High TcGLIP gene expression was observed in the leaves of mature plants and seedlings as well as in buds and flowers, a finding that was consistent with the pyrethrin I content in these parts. Expression was enhanced in response to wounding, which suggested that the enzyme plays a key role in the defense mechanism of T. cinerariifolium.

Links

PubMed Online version:10.1111/j.1365-313X.2012.04980.x

Keywords


Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

TANCI:GLIP

located_in

GO:0005615: extracellular space

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

TANCI:GLIP

involved_in

GO:0008299: isoprenoid biosynthetic process

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

TANCI:GLIP

involved_in

GO:0009611: response to wounding

ECO:0000270: expression pattern evidence used in manual assertion

P

Seeded From UniProt

complete

TANCI:GLIP

enables

GO:0016746: acyltransferase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

TANCI:GLIP

enables

GO:0016788: hydrolase activity, acting on ester bonds

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

TANCI:H6U1I8

enables

GO:0016412: serine O-acyltransferase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

TANCI:H6U1I8

GO:0016412: serine O-acyltransferase activity

ECO:0000315:

F

GDSL lipase was purified and recombineered into E. coli with a maltose binding protein fusion. The reaction transfers the chrysanthemoyl group from coenzyme A to pyrethrolone. Figure 3 shows mutations in serine of the GDSL domain eliminate acetyltransferase activity and maltose binding protein alone does not have acetyltransferase activity.

complete
CACAO 9773

See also

References

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