Molecular cloning and nucleotide sequences of cDNA and gene encoding endo-inulinase from Penicillium purpurogenum.
A cDNA and a gene encoding endo-inulinase from Penicillium purpurogenum were isolated, and were cloned for the first time. Two oligonucleotide probes, which were synthesized based on the partial amino acid sequences of the purified endo-inulinase, were used to screen a cDNA library. A 1.7-kb DNA fragment encoding endo-inulinase was isolated and analyzed. A single open reading frame, consisting of 1548-bp, was found to encode a polypeptide that comprised a 25-amino acid signal peptide and 490-amino acid mature protein. All the partial amino acid sequences of the purified enzyme were discovered in the deduced ones. The deduced amino acid sequences of endo-inulinase had similar sequences to those of fructan hydrolases. A 3.5-kb chromosomal DNA fragment encoding endo-inulinase was also isolated and analyzed. The same ORF with cDNA clone as identified. There were no introns in the endo-inulinase gene.
Onodera
S
S
Department of Food Science, Faculty of Dairy Science, Rakuno Gakuen University, Japan.
Murakami
T
T
Ito
H
H
Mori
H
H
Matsui
H
H
Honma
M
M
Chiba
S
S
Shiomi
N
N
D000595:Amino Acid Sequence; D001483:Base Sequence; D002241:Carbohydrates / Q000032:analysis; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics*; D015698:Genomic Library; D006026:Glycoside Hydrolases / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016366:Open Reading Frames; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics*; D010957:Plasmids
D002241:Carbohydrates; D018076:DNA, Complementary; D015345:Oligonucleotide Probes; D006026:Glycoside Hydrolases; C023086:inulinase