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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0657

PubMed

33208778, Sci Rep. 2020 Nov 18;10(1):20066. doi: 10.1038/s41598-020-77133-8.

Characterization method

recombinant protein expression,NMR

Genomic accession number

LC529899.1

Nucelotide position range

1-7951

Substrate

levoglucosan 

Loci

None

Species

Bacillus smithii S-2701M/1479

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

lgdB1 - BCB28820.1 0 - 822 (+) LC529899.1:1-823 -
- - BCB28821.1 849 - 2010 (+) LC529899.1:850-2011 -
lgdA - BCB28822.1 2058 - 3243 (+) LC529899.1:2059-3244 -
rbsA - BCB28823.1 3313 - 4801 (+) LC529899.1:3314-4802 -
- - BCB28824.1 4781 - 5180 (+) LC529899.1:4782-5181 -
- - BCB28825.1 5245 - 5785 (+) LC529899.1:5246-5786 -
lgdC - BCB28826.1 5807 - 6968 (+) LC529899.1:5808-6969 -
lgdB2 - BCB28827.1 6979 - 7951 (+) LC529899.1:6980-7952 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 822 (+) STP: STP|Peripla_BP_1 No
- 850 - 2010 (+) CDS No
- 2059 - 3243 (+) CAZyme: GH109 Yes
- 3314 - 4801 (+) TC: gnl|TC-DB|P0AGI1|3.A.1.2.1 Yes
- 4782 - 5180 (+) other Yes
- 5246 - 5785 (+) other Yes
- 5808 - 6968 (+) CAZyme: GH109 Yes
- 6980 - 7951 (+) CDS No

PUL ID

PUL0657

PubMed

33208778, Sci Rep. 2020 Nov 18;10(1):20066. doi: 10.1038/s41598-020-77133-8.

Title

Conversion of levoglucosan into glucose by the coordination of four enzymes through oxidation, elimination, hydration, and reduction.

Author

Kuritani Y, Sato K, Dohra H, Umemura S, Kitaoka M, Fushinobu S, Yoshida N

Abstract

Levoglucosan (LG) is an anhydrosugar produced through glucan pyrolysis and is widely found in nature. We previously isolated an LG-utilizing thermophile, Bacillus smithii S-2701M, and suggested that this bacterium may have a metabolic pathway from LG to glucose, initiated by LG dehydrogenase (LGDH). Here, we completely elucidated the metabolic pathway of LG involving three novel enzymes in addition to LGDH. In the S-2701M genome, three genes expected to be involved in the LG metabolism were found in the vicinity of the LGDH gene locus. These four genes including LGDH gene (lgdA, lgdB1, lgdB2, and lgdC) were expressed in Escherichia coli and purified to obtain functional recombinant proteins. Thin layer chromatography analyses of the reactions with the combination of the four enzymes elucidated the following metabolic pathway: LgdA (LGDH) catalyzes 3-dehydrogenation of LG to produce 3-keto-LG, which undergoes beta-elimination of 3-keto-LG by LgdB1, followed by hydration to produce 3-keto-D-glucose by LgdB2; next, LgdC reduces 3-keto-D-glucose to glucose. This sequential reaction mechanism resembles that proposed for an enzyme belonging to glycoside hydrolase family 4, and results in the observational hydrolysis of LG into glucose with coordination of the four enzymes.