title: Degradation of konjac glucomannan by Thermobifida fusca thermostable β-mannanase from yeast transformant.
abstract: Native konjac glucomannan was used as the substrate for thermophilic actinomycetes, Thermobifida fusca BCRC19214, to produce β-mannanase. The β-mannanase was purified and five internal amino acid sequences were determined by LC-MS/MS. These sequences had high homology with the β-mannanase from T. fusca YX. The tfm gene which encoded the β-mannanase was cloned, sequenced and heterologous expressed in Yarrowia lipolytica P01 g expression system. Recombinant heterologous expression resulted in extracellular β-mannanase production at levels as high as 3.16 U/ml in the culture broth within 48 h cultivation. The recombinant β-mannanase from Y. lipolytica transformant had superior thermal property. The optimal temperature of the recombinant β-mannanase from Y. lipolytica transformant (pYLSC1-tfm) was 80°C. When native konjac glucomannan was incubated with the recombinant β-mannanase from Y. lipolytica transformant (pYLSC1-tfm) at 50°C, there was a fast decrease of viscosity happen during the initial phase of reaction. This viscosity reduction was accompanied by an increase of reducing sugars. The surface of konjac glucomannan film became smooth. After 24h of treatment, the DPw of native konjac glucomannan decreased from 6,435,139 to 3089.
authors.affiliation: Department of Cosmetic Science, Providence University, Taichung 43301, Taiwan; Graduate Institute of Biotechnology, National Chung Hsing University, Taichung 40227, Taiwan.
authors.forename: Cheng-Yu
authors.initials: CY
authors.lastname: Chen
authors.affiliation: Department of Cosmetic Science, Providence University, Taichung 43301, Taiwan.
authors.forename: Yu-Chun
authors.initials: YC
authors.lastname: Huang
authors.affiliation: Department of Cosmetic Science, Providence University, Taichung 43301, Taiwan.
authors.forename: Ting-Ya
authors.initials: TY
authors.lastname: Yang
authors.affiliation: Department of Cosmetic Science, Providence University, Taichung 43301, Taiwan.
authors.forename: Jhen-Yi
authors.initials: JY
authors.lastname: Jian
authors.affiliation: Department of Cosmetic Science, Providence University, Taichung 43301, Taiwan; Department of Applied Chemistry, Providence University, Taichung 43301, Taiwan.
authors.forename: Wei-Lin
authors.initials: WL
authors.lastname: Chen
authors.affiliation: Department of Cosmetic Science, Providence University, Taichung 43301, Taiwan; Department of Applied Chemistry, Providence University, Taichung 43301, Taiwan. Electronic address: chyang@pu.edu.tw.
authors.forename: Chao-Hsun
authors.initials: CH
authors.lastname: Yang
mesh_terms: D004789:Enzyme Activation; D004795:Enzyme Stability; D005285:Fermentation; D006868:Hydrolysis; D008351:Mannans / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D015003:Yeasts / Q000201:enzymology* / Q000235:genetics; D044902:beta-Mannosidase / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D008351:Mannans; D011994:Recombinant Proteins; C022901:(1-6)-alpha-glucomannan; D044902:beta-Mannosidase
keywords: Heterologous expression; Konjac glucomannan; Thermobifida fusca; Yarrowia lipolytica; β-Mannanase