dbCAN-PUL

Because CGCFinder predicted no CGC for this PUL, the gene cluster depicted below contains dbCAN2 and CGC signature predictions for all genes in the PUL, instead of a predicted CGC.

PUL ID	PUL0174
PubMed	27302067, J Biol Chem. 2016 Aug 5;291(32):16438-47. doi: 10.1074/jbc.M116.727305. Epub 2016 Jun 14.
Characterization method	RT-PCR,enzyme activity assay,enzymatic product analysis
Genomic accession number	CP001736.1
Nucelotide position range	4338948-4346415
Substrate	starch
Loci	Kfla_4051-Kfla_4053
Species	Kribbella flavida/182640
Degradation or Biosynthesis	degradation

Gene Name	Locus Tag	Protein ID	Gene Position	GenBank Contig Range	EC Number
-	Kfla_4051	ADB33100.1	0 - 2847 (-)	CP001736.1:4338948-4341795	-
-	Kfla_4052	ADB33101.1	2843 - 6170 (-)	CP001736.1:4341791-4345118	-
-	Kfla_4053	ADB33102.1	6286 - 7468 (-)	CP001736.1:4345234-4346416	-

Cluster number	0
Gene name	Gene position	Gene type	Found by CGCFinder?
-	1 - 2847 (-)	CAZyme: CBM20\|GH31	No
-	2844 - 6170 (-)	CAZyme: CBM35\|GH31	No
-	6287 - 7468 (-)	TF: DBD-SUPERFAMILY\|0040266	No

PUL ID	PUL0174
PubMed	27302067, J Biol Chem. 2016 Aug 5;291(32):16438-47. doi: 10.1074/jbc.M116.727305. Epub 2016 Jun 14.
Title	Two Novel Glycoside Hydrolases Responsible for the Catabolism of Cyclobis-(1-->6)-alpha-nigerosyl.
Author	Tagami T, Miyano E, Sadahiro J, Okuyama M, Iwasaki T, Kimura A
Abstract	The actinobacterium Kribbella flavida NBRC 14399(T) produces cyclobis-(1-->6)-alpha-nigerosyl (CNN), a cyclic glucotetraose with alternate alpha-(1-->6)- and alpha-(1-->3)-glucosidic linkages, from starch in the culture medium. We identified gene clusters associated with the production and intracellular catabolism of CNN in the K. flavida genome. One cluster encodes 6-alpha-glucosyltransferase and 3-alpha-isomaltosyltransferase, which are known to coproduce CNN from starch. The other cluster contains four genes annotated as a transcriptional regulator, sugar transporter, glycoside hydrolase family (GH) 31 protein (Kfla1895), and GH15 protein (Kfla1896). Kfla1895 hydrolyzed the alpha-(1-->3)-glucosidic linkages of CNN and produced isomaltose via a possible linear tetrasaccharide. The initial rate of hydrolysis of CNN (11.6 s(-1)) was much higher than that of panose (0.242 s(-1)), and hydrolysis of isomaltotriose and nigerose was extremely low. Because Kfla1895 has a strong preference for the alpha-(1-->3)-isomaltosyl moiety and effectively hydrolyzes the alpha-(1-->3)-glucosidic linkage, it should be termed 1,3-alpha-isomaltosidase. Kfla1896 effectively hydrolyzed isomaltose with liberation of beta-glucose, but displayed low or no activity toward CNN and the general GH15 enzyme substrates such as maltose, soluble starch, or dextran. The kcat/Km for isomaltose (4.81 +/- 0.18 s(-1) mm(-1)) was 6.9- and 19-fold higher than those for panose and isomaltotriose, respectively. These results indicate that Kfla1896 is a new GH15 enzyme with high substrate specificity for isomaltose, suggesting the enzyme should be designated an isomaltose glucohydrolase. This is the first report to identify a starch-utilization pathway that proceeds via CNN.

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