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

文献类型： 外文期刊

第一作者： Wang, Xunzhang

作者： Wang, Xunzhang;Bei, Jinlong;Chen, Zhuang;Jiang, Zongyong

作者机构：

关键词： 6-Phytase;Chimeras;Enzymes;Nucleotide 6-Phytase;Chimeras;Enzymes;Nucleotide sequence;Protein engineering;pH effects;Aspergillus fumigatus;Aspergillus niger;Pichia pastoris

期刊名称：JOURNAL OF BIOTECHNOLOGY （ 影响因子：3.307； 五年影响因子：3.778 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： 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 alignmentwas 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 pH2.5. Most influences of substitutions were additive, except those of regions V and VI. Exchanging both regions led to different impacts upon K sub(m) and activity at pi H 4.0 compared with the replacement of either.

分类号： Q81

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