PMID- 8157587
OWN - NLM
STAT- MEDLINE
DCOM- 19940513
LR  - 20190508
IS  - 0021-9193 (Print)
IS  - 1098-5530 (Electronic)
IS  - 0021-9193 (Linking)
VI  - 176
IP  - 8
DP  - 1994 Apr
TI  - Cloning, sequencing, and disruption of a levanase gene of Bacillus polymyxa CF43.
PG  - 2177-83
AB  - The Bacillus polymyxa CF43 lelA gene, expressing both sucrose and fructan 
      hydrolase activities, was isolated from a genomic library of B. polymyxa screened 
      in Bacillus subtilis. The gene was detected as expressing sucrose hydrolase 
      activity; B. subtilis transformants did not secrete the lelA gene product (LelA) 
      into the extracellular medium. A 1.7-kb DNA fragment sufficient for lelA 
      expression in Escherichia coli was sequenced. It contains a 548-codon open 
      reading frame. The deduced amino acid sequence shows 54% identity with mature B. 
      subtilis levanase and is similar to other fructanases and sucrases 
      (beta-D-fructosyltransferases). Multiple-sequence alignment of 14 of these 
      proteins revealed several previously unreported features. LelA appears to be a 
      512-amino-acid polypeptide containing no canonical signal peptide. The hydrolytic 
      activities of LelA on sucrose, levan, and inulin were compared with those of B. 
      subtilis levanase and sucrase, confirming that LelA is indeed a fructanase. The 
      lelA gene in the chromosome of B. polymyxa was disrupted with a chloramphenicol 
      resistance gene (cat) by "inter-gramic" conjugation: the lelA::cat insertion on a 
      mobilizable plasmid was transferred from an E. coli transformant to B. polymyxa 
      CF43, and B. polymyxa transconjugants containing the lelA::cat construct 
      replacing the wild-type lelA gene in their chromosomes were selected directly. 
      The growth of the mutant strain on levan, inulin, and sucrose was not affected.
FAU - Bezzate, S
AU  - Bezzate S
AD  - Institut National de la Recherche Agronomique, Thiverval-Grignon, France.
FAU - Steinmetz, M
AU  - Steinmetz M
FAU - Aymerich, S
AU  - Aymerich S
LA  - eng
SI  - GENBANK/Z26651
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Bacteriol
JT  - Journal of bacteriology
JID - 2985120R
RN  - 0 (Bacterial Proteins)
RN  - 0 (DNA, Bacterial)
RN  - EC 3.2.1.- (Glycoside Hydrolases)
RN  - EC 3.2.1.65 (levanase)
SB  - IM
GS  - lelA
MH  - Amino Acid Sequence
MH  - Bacillus/*enzymology/genetics
MH  - Bacillus subtilis/genetics
MH  - *Bacterial Proteins
MH  - Base Sequence
MH  - Cloning, Molecular
MH  - Conjugation, Genetic/genetics
MH  - DNA, Bacterial/genetics
MH  - Escherichia coli/genetics
MH  - Genes, Bacterial/*genetics
MH  - Glycoside Hydrolases/*genetics/metabolism
MH  - Molecular Sequence Data
MH  - Sequence Homology, Amino Acid
PMC - PMC205337
EDAT- 1994/04/01 00:00
MHDA- 1994/04/01 00:01
PMCR- 1994/04/01
CRDT- 1994/04/01 00:00
PHST- 1994/04/01 00:00 [pubmed]
PHST- 1994/04/01 00:01 [medline]
PHST- 1994/04/01 00:00 [entrez]
PHST- 1994/04/01 00:00 [pmc-release]
AID - 10.1128/jb.176.8.2177-2183.1994 [doi]
PST - ppublish
SO  - J Bacteriol. 1994 Apr;176(8):2177-83. doi: 10.1128/jb.176.8.2177-2183.1994.