title: Combinatorial strategy of sorbitol feeding and low-temperature induction leads to high-level production of alkaline β-mannanase in Pichia pastoris.
abstract: A process for efficient production of an alkaline β-mannanases from Bacillus sp. N16-5 was established by heterologous expression using Pichia pastoris. A high producing strain was generated by removing the native β-mannanases signal peptide and increasing the copy number of the mature β-mannanases gene. High cell density fermentation of this strain in 1-L bioreactor led to a production level of 4164 U/mL after 96 h of induction. Sorbitol co-feeding and temperature-lowering strategies both increased the β-mannanase production levels. Combined usage of these two strategies achieved the most effective result-the enzyme level reached 6336 U/mL within 84 h, which to our best knowledge is the highest production level reported for the expression of extreme β-mannanase thus far. The strategy described in this work can also be adapted to express other important industrial enzymes with extreme properties.
authors: Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, PR China.
authors: Taicheng
authors: T
authors: Zhu
authors: Lijin
authors: L
authors: You
authors: Fuyu
authors: F
authors: Gong
authors: Minfeng
authors: M
authors: Xie
authors: Yanfen
authors: Y
authors: Xue
authors: Yin
authors: Y
authors: Li
authors: Yanhe
authors: Y
authors: Ma
mesh_terms: D001407:Bacillus / Q000201:enzymology* / Q000254:growth & development; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D003080:Cold Temperature; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D013012:Sorbitol / Q000378:metabolism*; D044902:beta-Mannosidase / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D013012:Sorbitol; D044902:beta-Mannosidase