title: Production of high activity Aspergillus niger BCC4525 β-mannanase in Pichia pastoris and its application for mannooligosaccharides production from biomass hydrolysis.
abstract: A cDNA encoding β-mannanase was cloned from Aspergillus niger BCC4525 and expressed in Pichia pastoris KM71. The secreted enzyme hydrolyzed locust bean gum substrate with very high activity (1625 U/mL) and a relatively high kcat/Km(461 mg-1s-1mL). The enzyme is thermophilic and thermostable with an optimal temperature of 70 °C and 40% retention of endo-β-1,4-mannanase activity after preincubation at 70 °C. In addition, the enzyme exhibited broad pH stability with an optimal pH of 5.5. The recombinant enzyme hydrolyzes low-cost biomass, including palm kernel meal (PKM) and copra meal, to produce mannooligosaccharides, which is used as prebiotics to promote the growth of beneficial microflora in animals. An in vitro digestibility test simulating the gastrointestinal tract system of broilers suggested that the recombinant β-mannanase could effectively liberate reducing sugars from PKM-containing diet. These characteristics render this enzyme suitable for utilization as a feed additive to improve animal performance.
authors.affiliation: a National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency , Pathum Thani , Thailand.
authors.forename: Piyanun
authors.initials: P
authors.lastname: Harnpicharnchai
authors.affiliation: b Faculty of Science, Department of Medical Sciences , Rangsit University , Pathum Thani , Thailand.
authors.forename: Waraporn
authors.initials: W
authors.lastname: Pinngoen
authors.affiliation: b Faculty of Science, Department of Medical Sciences , Rangsit University , Pathum Thani , Thailand.
authors.forename: Wanwisa
authors.initials: W
authors.lastname: Teanngam
authors.affiliation: a National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency , Pathum Thani , Thailand.
authors.forename: Warasirin
authors.initials: W
authors.lastname: Sornlake
authors.affiliation: a National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency , Pathum Thani , Thailand.
authors.forename: Kittapong
authors.initials: K
authors.lastname: Sae-Tang
authors.affiliation: b Faculty of Science, Department of Medical Sciences , Rangsit University , Pathum Thani , Thailand.
authors.forename: Pennapa
authors.initials: P
authors.lastname: Manitchotpisit
authors.affiliation: a National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency , Pathum Thani , Thailand.
authors.forename: Sutipa
authors.initials: S
authors.lastname: Tanapongpipat
mesh_terms: D000383:Agriculture; D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D018533:Biomass; D003001:Cloning, Molecular; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008358:Mannose / Q000737:chemistry; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry*; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D044902:beta-Mannosidase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*
chemical_list: D009844:Oligosaccharides; D011994:Recombinant Proteins; D044902:beta-Mannosidase; D008358:Mannose
keywords: copra meal; food and feed; mannanase; mannooligosaccharides; palm kernel meal