RRC ID 34054
Author Pudjiraharti S, Ohtani M, Takano N, Abe A, Lisdiyanti P, Tanaka M, Sone T, Asano K.
Title Nonomuraea sp. ID06-A0189 inulin fructotransferase (DFA III-forming): gene cloning, characterization and conservation among other Nonomuraea species.
Journal J. Antibiot.
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.
Volume 67(2)
Pages 137-41
Published 2014-2
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 2.033
Times Cited 4
WOS Category PHARMACOLOGY & PHARMACY BIOTECHNOLOGY & APPLIED MICROBIOLOGY IMMUNOLOGY MICROBIOLOGY
Resource
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