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

Paper ID Volume ID Publish Year Pages File Format Full-Text
17290 42657 2011 6 PDF Available
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.

► 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.

Keywords
Multi-copy; Pichia pastoris; Alkaline mannanase; High-level production; Co-feeding
First Page Preview
Combinatorial strategy of sorbitol feeding and low-temperature induction leads to high-level production of alkaline β-mannanase in Pichia pastoris
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 49, Issue 4, 10 September 2011, Pages 407–412
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering