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Structure-based fragment shuffling of two fungal phytases for combination of desirable properties

Paper ID Volume ID Publish Year Pages File Format Full-Text
24891 43543 2009 8 PDF Available
Title
Structure-based fragment shuffling of two fungal phytases for combination of desirable properties
Abstract

Aspergillus niger NRRL 3135 phytase (Anp) and Aspergillus fumigatus ATCC 13073 phytase (Afp) are quite different but mutually complementary in many properties. A semi-rational protein engineering strategy based on 3D structure and sequence alignment was used to take advantage of the desirable characteristics of both enzymes. Each phytase was divided into seven fragments, including regions I–VII (I, 1–47; II, 59–133; III, 139–172; IV, 178–237; V, 246–329; VI338–381; VII, 404–444). The equivalent regions were swapped to construct an array of chimeras. Among the functional chimeras expressed in the yeast Pichia pastoris, novel phytases with combinations of the most desirable properties, including heat-resistance, were obtained. Correlations of individual regions with detailed differences were established by systematic evaluation of the substitutions. Regions II and VI contributed to the difference in specific activity at pH 5.0. Regions IV and V of Anp fully accounted for its second pH optimum at pH 2.5. Most influences of substitutions were additive, except those of regions V and VI. Exchanging both regions led to different impacts upon Km and activity at ∼pH 4.0 compared with the replacement of either.

Keywords
Aspergillus fumigatus; Aspergillus niger; Phytase; Protein engineering; Heat-resistance
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Structure-based fragment shuffling of two fungal phytases for combination of desirable properties
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 139, Issue 2, 15 January 2009, Pages 186–193
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us