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Purification and characterization of a β-1,3-glucomannanase expressed in Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
17410 42666 2011 6 PDF Available
Title
Purification and characterization of a β-1,3-glucomannanase expressed in Pichia pastoris
Abstract

The glycoside hydrolase β-1,3-glucomannanase is an enzyme that specifically breaks the β-1,3 glycosidic bond of the glucomannan, the main cell wall constituent of some yeasts. In this work, a codon optimized DNA sequence of the MAN5C gene from Penicillium lilacinum ATCC 36010 was expressed in the yeast Pichia pastoris under the control of AOX1 promoter. The recombinant protein plMAN5C was purified from the shake flask culture and the stirred-tank bioreactor culture in yields of 30.0 mg/l and 224.0 mg/l, respectively. The purified protein had a specific activity of 14.6 U/mg at 37 °C, pH 4.5. Biochemical analysis showed that the optimal temperature and pH for plMAN5C were 50 °C and 4.5, respectively. The recombinant plMAN5C was efficient in lysis of the cell wall of the red yeast Rhodosporidium toruloides to form protoplast. Our work provided an effective system for heterogeneous production of β-1,3-glucomannanase, which should facilitate a more convenient application of this enzyme in biotechnology and other related areas.

Keywords
β-1,3-glucomannanase; Secretory expression; Pichia pastoris; Cell wall; Enzyme activity
First Page Preview
Purification and characterization of a β-1,3-glucomannanase expressed in Pichia pastoris
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 49, Issue 2, 10 July 2011, Pages 223–228
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering