title: Genome mining and motif truncation of glycoside hydrolase family 5 endo-β-1,4-mannanase encoded by Aspergillus oryzae RIB40 for potential konjac flour hydrolysis or feed additive.
abstract: Two novel glycosyl hydrolase family 5 (GH5) β-mannanases (AoMan5A and AoMan5B) were identified from Aspergillus oryzae RIB40 by genome mining. The AoMan5A contains a predicted family 1 carbohydrate binding module (CBM-1), located at its N-terminal. The AoMan5A, AoMan5B and truncated mutant AoMan5AΔCL (truncating the N-terminal CBM and linker of AoMan5A) were expressed retaining the N-terminus of the native protein in Pichia pastoris GS115 by pPIC9KM. The specific enzyme activity of the purified reAoMan5A, reAoMan5B and reAoMan5AΔCL towards locust bean gum at pH 3.6 and 40°C for 10min, was 8.3, 104.2 and 15.8U/mg, respectively. The temperature properties of the reAoMan5AΔCL were improved by truncating CBM. They can degrade the pretreated konjac flour and produce prebiotics. In addition, they had excellent stability under simulative gastric fluid and simulative prilling process. All these properties make these recombinant β-mannanases potential additives for use in the food and feed industries.
authors.affiliation: Henan Provincial Engineering Laboratory of Insect Bio-reactor, Nanyang Normal University, 1638 Wolong Road, Nanyang, Henan 473061, People's Republic of China.
authors.forename: Cun-Duo
authors.initials: CD
authors.lastname: Tang
authors.affiliation: Henan Provincial Engineering Laboratory of Insect Bio-reactor, Nanyang Normal University, 1638 Wolong Road, Nanyang, Henan 473061, People's Republic of China.
authors.forename: Hong-Ling
authors.initials: HL
authors.lastname: Shi
authors.affiliation: Henan Provincial Engineering Laboratory of Insect Bio-reactor, Nanyang Normal University, 1638 Wolong Road, Nanyang, Henan 473061, People's Republic of China.
authors.forename: Qing-Hai
authors.initials: QH
authors.lastname: Tang
authors.affiliation: Henan Provincial Engineering Laboratory of Insect Bio-reactor, Nanyang Normal University, 1638 Wolong Road, Nanyang, Henan 473061, People's Republic of China.
authors.forename: Jun-Shi
authors.initials: JS
authors.lastname: Zhou
authors.affiliation: Henan Provincial Engineering Laboratory of Insect Bio-reactor, Nanyang Normal University, 1638 Wolong Road, Nanyang, Henan 473061, People's Republic of China. Electronic address: lunguangyao@163.com.
authors.forename: Lun-Guang
authors.initials: LG
authors.lastname: Yao
authors.affiliation: Henan Provincial Engineering Laboratory of Insect Bio-reactor, Nanyang Normal University, 1638 Wolong Road, Nanyang, Henan 473061, People's Republic of China.
authors.forename: Zhu-Jin
authors.initials: ZJ
authors.lastname: Jiao
authors.affiliation: Henan Provincial Engineering Laboratory of Insect Bio-reactor, Nanyang Normal University, 1638 Wolong Road, Nanyang, Henan 473061, People's Republic of China. Electronic address: kanyunchao@163.com.
authors.forename: Yun-Chao
authors.initials: YC
authors.lastname: Kan
mesh_terms: D020816:Amino Acid Motifs; D031056:Amorphophallus; D000821:Animal Feed; D000818:Animals; D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D004795:Enzyme Stability; D005503:Food Additives; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D005685:Galactans; D016681:Genome, Fungal; D006868:Hydrolysis; D008351:Mannans / Q000378:metabolism; D008361:Mannosidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010843:Pichia / Q000235:genetics; D053149:Plant Gums; D056692:Prebiotics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D005503:Food Additives; D005656:Fungal Proteins; D005685:Galactans; D008351:Mannans; D053149:Plant Gums; D056692:Prebiotics; D011994:Recombinant Proteins; C022901:(1-6)-alpha-glucomannan; D008361:Mannosidases; C041867:endo-1,4-beta-D-mannanase; C017471:locust bean gum
keywords: Aspergillus oryzae; Carbohydrate binding module; Genome mining; Pichia pastoris; β-mannanase