Abstract
Xylanase gene mutation by error-prone pcr and expression in Pichia pastoris
Highlights
Xylan is a major component of hemicellulose, which widely presents in the plant cell wall and is a big renewable resource in nature (COLLINS et al, 2005)
The supernatant of the recombinant P. pastoris with xylanase gene was performed by boiling at 100°C for 5 min with equal volume of 5×Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) loading buffer, and centrifuged at 10000 rounds per min (RPM) for 3 min
The mutant xylanase genes were obtained by error-prone PCR amplification
Summary
Xylan is a major component of hemicellulose, which widely presents in the plant cell wall and is a big renewable resource in nature (COLLINS et al, 2005). Error-prone PCR causes mutation of gene with less than 800 bp, which can change the natural parameters of the native enzymes. It was reported that two mutant protease genes of Bacillus subtilis were obtained by error-prone PCR, their catalytic efficiency was respectively 256 and 131 folds higher than the native enzyme (CHEN and ARNOLD, 1991 and 1993).
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