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

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 mannooligosaccharides to release mannose and mannobiose. rGal27A showed synergistic interactions with rMan113A and recombinant β-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.

A Recombinant β-Mannanase from Thermoanaerobacterium aotearoense SCUT27: Biochemical Characterization and Its Thermostability Improvement

“…CFR1601 (GH26), and Man113A from Bacillus sp. N16-5 . Enzymes with an optimal activity and thermostability at high temperature are valuable for industrial applications.…”

“…β-Mannanase is available from many sources, including bacteria, fungi, actinomycetes, plants, and animals, − and can be classified into different glycoside hydrolase (GH) families (e.g., 5, 26, and 113 ,, ) according to the Carbohydrate-Active Enzymes database ( ). The families GH5, GH26, and GH113 belong to clan GH-A, and they share a similar protein folding pattern and catalytic mechanism. , Some mannanases possess an additional carbohydrate binding module (CBM), which could provide thermo-protection to the GH domain or alter the substrate specificity of the enzyme. , …”

“…6 An enzyme mixture containing β-mannanase prepared from Aspergillus aculeatus dramatically reduced the viscosity of cassava, and the subsequent mash was applied for very high gravity fermentation by yeasts. 7,8 β-Mannanase is available from many sources, including bacteria, fungi, actinomycetes, plants, and animals, 9−11 and can be classified into different glycoside hydrolase (GH) families (e.g., 5, 26, and 113 2,12,13 ) according to the Carbohydrate-Active Enzymes database (www.cazy.org 14 ). The families GH5, GH26, and GH113 belong to clan GH-A, and they share a similar protein folding pattern and catalytic mechanism.…”

Bagasse minority pathway expression: Real time study of GH2 β-mannosidases from bacteroidetes

“…In addition, the literature has recently reported the importance of regulating systems similar to PULs in other phyla, such as the mechanism related to the use of mannan in Bacillus sp. in Firmicutes [60]. Many of the PUL pathway enzymes are responsible for the natural deconstruction of biomass in order to acquire nutrition from decomposing plant material.…”

Recombinant production and characterization of six novel GH27 and GH36 α-galactosidases from Penicillium subrubescens and their synergism with a commercial mannanase during the hydrolysis of lignocellulosic biomass

“…These differences can be explained by the higher extent of galactose substitutions on the mannan backbone of guar gum, which makes the debranching of the substrate by the α-galactosidases more critical (Aulitto et al, 2018;Song et al, 2018). Notably, AglB alone achieved about 90% of relative galactose release, while the combination with endomannanase produced the highest sugar release (about 100%), but the higher release of galactose did not correspond with a high DS value, 1.08 (Table 3).…”