| RRC ID |
34054
|
| 著者 |
Pudjiraharti S, Ohtani M, Takano N, Abe A, Lisdiyanti P, Tanaka M, Sone T, Asano K.
|
| タイトル |
Nonomuraea sp. ID06-A0189 inulin fructotransferase (DFA III-forming): gene cloning, characterization and conservation among other Nonomuraea species.
|
| ジャーナル |
J Antibiot (Tokyo)
|
| Abstract |
The inulin fructotransferase (DFA III-forming)(EC 4.2.2.18) gene in Nonomuraea sp. ID06-A0189 was amplified from genomic DNA, sequenced and expressed in Escherichia coli. The 1326-bp gene, designated as Nsp-ift, encodes a protein composed of a putative 37-amino-acid signal peptide and 404-amino-acid mature protein. A putative ribosomal binding sequence was identified 12 bases upstream from the start codon. However, a typical bacterial promoter could not be found by in silico analysis. The deduced amino-acid sequence of the enzyme was most similar to that of inulin fructotransferase (DFA I-forming) in Frankia sp. EAN1pec. Phylogenetic analysis of deduced amino-acid sequences indicated that Nonomuraea sp. ID06-A0189 and Frankia sp. EAN1pec inulin fructotransferases formed a distinct clade from those from Arthrobacter sp. H65-7, A. globiformis and Bacillus sp. snu-7 that showed 57, 56 and 56% identity to that of Nsp-ift, respectively. The Nsp-ift without a putative signal peptide was successfully expressed in E. coli and partially purified using His-tag affinity chromatography. The recombinant enzyme displayed optimum temperature between 65 and 70 °C, optimum pH between 5.5 and 6.0 and remained stable up to 70 °C. The properties were identical to those of the original enzyme. Of 10 Nonomuraea species tested by Southern hybridization, enzyme activity measurements and PCR, only Nonomuraea sp. ID06-A0189 has the Nsp-ift gene, suggesting that Nsp-ift is not highly conserved in this genus.
|
| 巻・号 |
67(2)
|
| ページ |
137-41
|
| 公開日 |
2014-2-1
|
| DOI |
10.1038/ja.2013.95
|
| PII |
ja201395
|
| PMID |
24129686
|
| MeSH |
Actinomycetales / enzymology*
Actinomycetales / genetics
Amino Acid Sequence
Bacterial Proteins / classification*
Bacterial Proteins / genetics
Bacterial Proteins / metabolism*
Catalysis
Cloning, Molecular
Conserved Sequence
DNA, Bacterial / genetics
Escherichia coli / genetics
Escherichia coli / metabolism
Genes, Bacterial / genetics
Hexosyltransferases / chemistry*
Hexosyltransferases / classification*
Hexosyltransferases / genetics
Phylogeny
Promoter Regions, Genetic
Sequence Analysis, DNA
|
| IF |
2.668
|
| 引用数 |
6
|
|
WOS 分野
|
PHARMACOLOGY & PHARMACY
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
IMMUNOLOGY
MICROBIOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM 3109
JCM 3144
JCM 3145
JCM 3234
JCM 3283
JCM 3321
JCM 6834
JCM 9946
JCM 11136 |
