Cloning and characterization of the gene encoding phosphoketolase in Leuconostoc mesenteroides isolated from kimchi.
The gene encoding phosphoketolase, which is 2749 bp long and contains 814 amino acid polypeptides with a total molecular mass of 91.9 kDa, was cloned from Leuconostoc mesenteroides C7 (LMC7) and expressed in Escherichia coli. It exhibited a homology of >58% with phosphoketolases from other lactic acid bacteria. The phosphoketolase of LMC7 belongs to the xylulose 5-phosphate (X5P)/fructose 6-phosphate (F6P) phosphoketolase (Xfp) family, which is an enzyme with dual specificity for X5P and F6P. The members of this family contain typical thiamin pyrophosphate (TPP) binding sites as reported for other TPP-dependent enzymes, and several highly conserved regions as signature patterns for phosphoketolases. The plasmid pGPK containing the Xfp gene (xfp) exhibits phosphoketolase activity in E. coli. The specific activities of the enzyme from E. coli BL21 and E. coli EC101 harboring xfp were 0.28 and 0.14 units/mg, respectively. They both exhibited a 1.5-fold increase in the production of acetic acid from acetyl phosphate compared with their corresponding original strain.
Lee
Jung Min
JM
School of Agricultural Biotechnology, Seoul National University, Republic of Korea.
Jeong
Do-Won
DW
Koo
Ok Kyung
OK
Kim
Ok Kyung Min Jung
OK
Kim
Min Jung
MJ
Lee
Jong-Hoon
JH
Chang
Hae Choon
HC
Kim
Jeong Hwan
JH
Lee
Hyong Joo
HJ
D000446:Aldehyde-Lyases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005636:Fructosephosphates / Q000378:metabolism; D007934:Leuconostoc / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010428:Pentosephosphates / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D004269:DNA, Bacterial; D005636:Fructosephosphates; D010428:Pentosephosphates; C031625:xylulose-5-phosphate; C027618:fructose-6-phosphate; D000446:Aldehyde-Lyases; C022367:phosphoketolase