Expression and characterisation of neopullulanase from Lactobacillus mucosae.
OBJECTIVE
To clone and express a neopullulanase gene from Lactobacillus mucosae LM1 in Escherichia coli and characterise the resulting recombinant neopullulanase.


RESULTS
An ORF in L. mucosae corresponding to a neopullulanase was cloned and expressed in E. coli. The predicted amino acid sequence of the neopullulanase contained catalytic sites and conserved motifs that are present in members of the neopullulanase subfamily. The resulting recombinant neopullulanase was efficiently purified by Ni-NTA affinity chromatography. The purified enzyme optimally hydrolyses pullulan at 37 °C and pH 6.0, producing panose as the major reaction product.


CONCLUSIONS
To the best of our knowledge, this is the first report of the cloning, expression and characterisation of a neopullulanase gene from a lactic acid bacterium.
Department of Animal Resource Science, Dankook University, Cheonan, 31116, Republic of Korea.
Marilen P
MP
Balolong
Department of Animal Resource Science, Dankook University, Cheonan, 31116, Republic of Korea.
Jong Pyo
JP
Chae
Department of Animal Resource Science, Dankook University, Cheonan, 31116, Republic of Korea. dkkang@dankook.ac.kr.
Dae-Kyung
http://orcid.org/0000-0001-9241-1250
DK
Kang
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D003001:Cloning, Molecular / Q000379:methods*; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005936:Glucans / Q000096:biosynthesis* / Q000737:chemistry*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006868:Hydrolysis; D007778:Lactobacillus / Q000201:enzymology* / Q000235:genetics; D016366:Open Reading Frames; D013379:Substrate Specificity
D001426:Bacterial Proteins; D005936:Glucans; C008763:panose; C009109:pullulan; D006026:Glycoside Hydrolases; C058484:neopullulanase
Lactobacillus mucosae; Neopullulanase; Panose