Functional investigation of a gene encoding pteridine glycosyltransferase for cyanopterin synthesis in Synechocystis sp. PCC 6803.
A gene (slr1166) putatively encoding pteridine glycosyltransferase was disrupted with a kanamycin resistance cassette in Synechocystis sp. PCC 6803, which produces cyanopterin. The deduced polypeptide from slr1166 consisted of 354 amino acid residues sharing 45% sequence identity with UDP-glucose:tetrahydrobiopterin alpha-glucosyltransferase (BGluT) isolated previously from Synechococcus sp. PCC 7942. The knockout mutant was unable to produce cyanopterin but only 6-hydroxymethylpterin-beta-galactoside, verifying that slr1166 encodes a pteridine glycosyltransferase, which is responsible for transfer of the second sugar glucuronic acid in cyanopterin synthesis. The mutant was affected in its intracellular pteridine content and growth rate, which were 74% and 80%, respectively, of wild type, demonstrating that the second sugar residue is still required for quantitative maintenance of cyanopterin. This supports the previous suggestion that glycosylation may contribute to high cellular concentration of pteridine compounds.
Hwang
Yoon Kyung
YK
Department of Microbiology, Inje University, Kimhae 621-749, South Korea.
Kang
Ji Youn
JY
Woo
Hyun Joo
HJ
Choi
Yong Kee
YK
Park
Young Shik
YS
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D000458:Cyanobacteria / Q000201:enzymology / Q000235:genetics*; D004187:Disaccharides / Q000096:biosynthesis*; D005798:Genes, Bacterial; D005964:Glucosyltransferases / Q000235:genetics; D020723:Glucuronic Acid / Q000378:metabolism; D016695:Glycosyltransferases / Q000235:genetics* / Q000378:metabolism; D007613:Kanamycin Resistance / Q000235:genetics; D008969:Molecular Sequence Data; D011621:Pteridines / Q000378:metabolism; D011622:Pterins
D001426:Bacterial Proteins; D004187:Disaccharides; D011621:Pteridines; D011622:Pterins; C458439:cyanopterin; D020723:Glucuronic Acid; D016695:Glycosyltransferases; C437230:BGluT protein, Synechococcus sp. PCC 7942; D005964:Glucosyltransferases