Purification and functional characterization of endo-β-mannanase MAN5 and its application in oligosaccharide production from konjac flour

Abstract

MAN5, the main extracellular saccharide hydrolase from Bacillus sp. MSJ-5, is an endo-beta-mannanase with a demand of at least five sugar moieties for effective cleavage. It has a pH optimum of 5.5 and a temperature optimum of 50 degrees C and is stable at pH 5-9 or below 65 degrees C. MAN5 has a very high ability to hydrolyze konjac flour, 10 U/mg of which could completely liquefy konjac flour gum in 10 min at 50 degrees C. HPLC analysis showed that most glucomannan in the konjac flour was hydrolyzed into a large amount of oligosaccharides with DP of 2-6 and a very small amount of monosaccharide. With the culture supernatant as enzyme source, the optimum condition to prepare oligosaccharides from konjac flour was obtained as 10 mg/ml konjac flour incubated with 10 U/mg enzyme at 50 degrees C for 24 h. With this condition, more than 90% polysaccharides in the konjac flour solution were hydrolyzed into oligosaccharides and a little monosaccharide (2.98% of the oligosaccharides). Konjac flour is an underutilized agricultural material with low commercial value in China. With MAN5, konjac flour can be utilized to generate high value-added oligosaccharides. The high effectiveness and cheapness of this technique indicates its potential in industry.

Enzymatic Hydrolysis of Porang by Streptomyces violascens BF 3.10 Mannanase for the Production of Mannooligosaccharides

“…Recent studies reported that mannanase from certain bacteria can hydrolyze mannan into certain oligosaccharides, such as mannooligosaccharide production from palm kernel by mannanase of Brevibacillus borstelensis (Utami et al, 2013), also from copra pulp by recombinant of A. niger (Cuong et al, 2013), and from locust bean gum by using immobilized Penicillium occitanis mannanase (Blibech et al, 2011). Hydrolysis of porang glucomannan by mannanase of S. violascens BF 3.10 in this study produced the same products compared to hydrolysis of konjac flour by endo-β-mannanase MAN5 (Zhang et al, 2009). More than 90% polysaccharides were hydrolyzed by endo-β-mannanase MAN5 into oligosaccharides with DP 2-6 (Zhang et al, 2009).…”

“…Hydrolysis of porang glucomannan by mannanase of S. violascens BF 3.10 in this study produced the same products compared to hydrolysis of konjac flour by endo-β-mannanase MAN5 (Zhang et al, 2009). More than 90% polysaccharides were hydrolyzed by endo-β-mannanase MAN5 into oligosaccharides with DP 2-6 (Zhang et al, 2009). Some fungal mannanase hydrolyze mannotetraose to mannotriose and mannobiose through transglycosylation reaction (Puchart et al, 2004).…”

“…Konjac flour hydrolyzed by mannanase of Bacillus sp. MSJ-5 produced mannooligosaccharides with DP 2-6 (Zhang et al, 2009).…”

An endo-β-1,4-mannanase, AkMan, from the common sea hare Aplysia kurodai

“…Such inhibitory effects of Fe 3+ and Ag + on -1,4-mannanase have been reported in -1,4-mannanse from Bacillus sp. MSJ-5 (Zhang et al, 2009). Complete inactivation by Ag + and 40-50% inactivation by Co 2+ , Fe 2+ and Cu 2+ were reported in Haliotis, Pomacea and Littorina enzymes (Ootsuka et al, 2006;Yamaura and Matsumoto, 1993;Yamaura et al, 1996).…”

“…Tolerance to wide pH range was also reported in mannanase from Bacillus sp. MSJ-5 (Zhang et al, 2009) and Bacillus subtilis NM-39 (Mendoza et al, 1994).…”