PUL ID

PUL0030

PubMed

26978267, PLoS One. 2016 Mar 15;11(3):e0150059. doi: 10.1371/journal.pone.0150059. eCollection 2016.
30351049, J Agric Food Chem. 2018 Oct 24;66(42):11055-11063. doi: 10.1021/acs.jafc.8b03878. Epub 2018 Oct 15.

Characterization method

isothermal calorimetric titration,gene deletion mutant and growth assay,enzyme activity assay

Genomic accession number

KU644713.1

Nucelotide position range

1-16598

Substrate

galactomannan

Loci

manDREHKCIJBGFA

Species

Bacillus sp. N16-5/122631

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

- - AML27054.1 0 - 1020 (-) KU644713.1:1-1021 -
- - AML27055.1 1299 - 2304 (-) KU644713.1:1300-2305 -
- - AML27056.1 2365 - 3541 (-) KU644713.1:2366-3542 -
- - AML27057.1 3776 - 5084 (-) KU644713.1:3777-5085 -
- - AML27058.1 5854 - 7060 (-) KU644713.1:5855-7061 -
- - AML27059.1 8026 - 9196 (-) KU644713.1:8027-9197 -
- - AML27060.1 9549 - 10353 (-) KU644713.1:9550-10354 -
- - AML27061.1 10355 - 11297 (-) KU644713.1:10356-11298 -
- - AML27062.1 11363 - 12308 (-) KU644713.1:11364-12309 -
- - AML27063.1 12329 - 13544 (-) KU644713.1:12330-13545 -
- - AML27064.1 13791 - 14748 (-) KU644713.1:13792-14749 -
- - AML27065.1 15116 - 16598 (+) KU644713.1:15117-16599 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 1020 (-) CAZyme: GH130 Yes
- 1300 - 2304 (-) TF: DBD-Pfam|LacI,DBD-SUPERFAMILY|0036955 Yes
- 2366 - 3541 (-) CAZyme: GH130 Yes
- 3777 - 5084 (-) STP: STP|SBP_bac_1 Yes
- 5855 - 7060 (-) other Yes
- 8027 - 9196 (-) CAZyme: GH27 Yes
- 9550 - 10353 (-) TC: gnl|TC-DB|Q8DT26|3.A.1.1.27 Yes
- 10356 - 11297 (-) TC: gnl|TC-DB|P94529|3.A.1.1.34 Yes
- 11364 - 12308 (-) other Yes
- 12330 - 13544 (-) other Yes
- 13792 - 14748 (-) CAZyme: CE7 Yes
- 15117 - 16598 (+) CDS No

PUL ID

PUL0030

PubMed

26978267, PLoS One. 2016 Mar 15;11(3):e0150059. doi: 10.1371/journal.pone.0150059. eCollection 2016.

Title

A Novel Manno-Oligosaccharide Binding Protein Identified in Alkaliphilic Bacillus sp. N16-5 Is Involved in Mannan Utilization.

Author

Song Y, Li J, Meng S, Yin L, Xue Y, Ma Y

Abstract

ManH, a novel substrate-binding protein of an ABC transporter, was identified from the mannan utilization gene cluster of Bacillus sp. N16-5. We cloned, overexpressed, and purified ManH and measured its binding affinity to different substrates by isothermal titration calorimetry. ManH binds to mannotriose, mannotetraose, mannopentose, and galactosyl-mannotriose with dissociation constants in the micromolar range. Deletion of manH led to decreased growth ability of the strain when cultivated in medium with manno-oligosaccharides or mannan as the carbon source. ManH belongs to a manno-oligosaccharide transporter and plays an important role in mannan utilization by Bacillus sp. N16-5.

PubMed

30351049, J Agric Food Chem. 2018 Oct 24;66(42):11055-11063. doi: 10.1021/acs.jafc.8b03878. Epub 2018 Oct 15.

Title

Galactomannan Degrading Enzymes from the Mannan Utilization Gene Cluster of Alkaliphilic Bacillus sp. N16-5 and Their Synergy on Galactomannan Degradation.

Author

Song Y, Sun W, Fan Y, Xue Y, Liu D, Ma C, Liu W, Mosher W, Luo X, Li Z, Ma W, Zhang T

Abstract

Two glycoside hydrolases encoded by the mannan utilization gene cluster of alkaliphilic Bacillus sp. N16-5 were studied. The recombinant Gal27A (rGal27A) hydrolyzed both galactomannans and oligo-galactomannans to release galactose, while the recombinant Man113A (rMan113A) showed poor activity toward galactomannans, but it hydrolyzed manno-oligosaccharides to release mannose and mannobiose. rGal27A showed synergistic interactions with rMan113A and recombinant beta-mannanase ManA (rManA), which is also from Bacillus sp. N16-5, in galactomannan degradation. The synergy degree of rGal27A and rManA on hydrolysis of locust bean gum and guar gum was 1.13 and 2.21, respectively, and that of rGal27A and rMan113A reached 2.00 and 2.68. The main products of galactomannan hydrolyzed by rGal27A and rManA simultaneously were galactose, mannose, mannobiose, and mannotriose, while those of galactomannan hydrolyzed by rGal27A and rMan113A were galactose and mannose. The yields of mannose, mannobiose, and mannotriose dramatically increased compared with the hydrolysis in the presence of rManA or rMan113A alone.