Purification and Characterization of a Novel Exo-β-Mannanase from Aeromonas sp. F-25

Abstract

A novel exo-beta-mannanase (1,4-beta-D-mannan mannobiohydrolase) was isolated from the culture fluid of strain No. F-25 of Aeromonas hydrophila subspecies anaerogenes, and purified about 4,000-fold by ammonium sulfate precipitation and successive co.umn chromatographies. The final enzyme preparation appeared to be homogeneous on polyacrylamide gel electrophoresis. The enzyme hydrolyzed the beta-1,4-mannan link in polysaccharides of three or more beta-1,4-linked D-mannose units. The enzyme had a molecular weight of 64,000, pI of 5.9, pH optimum of 6.0, and was stable in a pH region of 5.0 to 8.5 and at temperatures below 45 degrees C. The Km values of the enzyme were 5.1 X 10(-4) M for mannotriose, 2.4 X 10(-4) M for mannotetraose and 1.3 X 10(-4) M for mannopentaose. The enzyme attacked codium and coffee mannans to give only mannobiose. Mannobiosyl- and mannotetraosyl-mannitol were hydrolyzed to produce mannobiose and mannitol, while mannobiose and mannosylmannitol were released from mannotriosylmannitol. The enzyme did not act on mannobiose, p-nitrophenyl-beta-D-mannoside, konjac glucomannan, or guar gum galactomannan. Furthermore, the enzyme catalyzed a transglycosylation reaction.

Cloning and Characterization of a β-1,4-Mannanase 5C Possessing a Family 27 Carbohydrate-Binding Module from a Marine Bacterium,Vibriosp. Strain MA-138

“…strain MA-138 (ManA) hydrolyzed mannooligosaccharides larger than mannotriose, producing several oligosaccharides. 11,[42][43][44][45] Man5C hydrolyzed mannooligosaccharides larger than mannotriose, producing several oligosaccharides, but did not hydrolyze mannotriose, mannobiose, or p-nitrophenyl--D-mannoside. Thus the enzyme was ascertained to be similar to endo--mannanases of the latter group.…”

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

“…-1,4-Mannanase has been isolated from bacteria (Araki, 1983 ;Akino et al, 1988;Talbot et al, 1990;Braithwaite et al, 1995;Nakajima and Matsuura, 1997;Li et al, 2000;Politz et al, 2000;Kansoh and Nagieb, 2004;Li et al, 2006), fungi (Johnson, 1990;Stalbrand et al, 1993;Kurakake et al, 2001;Puchart et al, 2004;Naganagouda et al, 2009), higher plants (Shimahara et al, 1975;Marraccini et al, 2001), and mollusks (Yamaura and Matsumoto, 1993;Yamaura et al, 1996;Xu et al, 2002a andOotsuka et al, 2006). Compared with bacterial and fungal enzymes, molluscan enzymes have not been well investigated, i.e., only four -1,4-mannanases have been investigated in Pomacea insularus (Yamaura and Matsumoto, 1993), Littorina brevicula (Yamaura et al, 1996), Mytilus edulis (Xu et al, 2002a and b), and Haliotis discus hannai (Ootsuka et.…”

Heterologous expression and characterization of man gene from Bacillus Subtilis in Pichia Pastoris

“…The b-Mannanase was first found in plants and germinating seeds, further it was isolated and identified in fungi, bacteria, even blue mussel (Shimahara et al, 1975;Araki, 1982;Akino et al, 1988;Johnson, 1990;Oda et al, 1993;Stålbrand et al, 1993;Xu et al, 2002a;Ootsuka et al, 2006). Because the data were available from several difficult sources on purification and characterization of b-1,4-mannanases, the recombinant enzymes were still poorly produced and expressed, and the expression level was relatively low (Stålbrand et al, 1993;Mendoza et al, 1995;Tang et al, 2001;Xu et al, 2002b;Mingardon et al, 2005).…”