Details

Autor(en) / Beteiligte

• Zhu, Taicheng
• You, Lijin
• Gong, Fuyu
• Xie, Minfeng
• Xue, Yanfen
• Li, Yin
• Ma, Yanhe

Titel

Combinatorial strategy of sorbitol feeding and low-temperature induction leads to high-level production of alkaline β-mannanase in Pichia pastoris

Ist Teil von

• Enzyme and microbial technology, 2011-09, Vol.49 (4), p.407-412

Ort / Verlag

Amsterdam: Elsevier Inc

Erscheinungsjahr

2011

Quelle

MEDLINE

Beschreibungen/Notizen

• ► Removal of the native signal peptide is a prerequisite to achieve high yield. ► Increasing the gene dosage dramatically increased the mannanase production level. ► The combinatorial strategy clearly led to better performance than either alone. ► The enzyme activity of the expressed alkaline β-mannanase reached 6336 U/ml. ► This is the highest alkaline β-mannanases level produced to date. 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.