5NFG

Structure of recombinant cardosin B from Cynara cardunculus

5NFG の概要

• エントリーDOI: 10.2210/pdb5nfg/pdb
• 分子名称: Procardosin-B,Procardosin-B, alpha-D-mannopyranose-(1-2)-[alpha-D-mannopyranose-(1-6)]alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, alpha-D-mannopyranose-(1-6)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, ... (4 entities in total)
• 機能のキーワード: plant aspartic protease, hydrolase
• 由来する生物種: Cynara cardunculus (Cardoon); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 86013.07

構造登録者

Pereira, P.J.B.,Figueiredo, A.C.,Manso, J.A.,Almeida, C.M.,Simoes, I. (登録日: 2017-03-14, 公開日: 2017-10-11, 最終更新日: 2024-11-06)

主引用文献

Almeida, C.M.,Manso, J.A.,Figueiredo, A.C.,Antunes, L.,Cruz, R.,Manadas, B.,Bur, D.,Pereira, P.J.B.,Faro, C.,Simoes, I.
Functional and structural characterization of synthetic cardosin B-derived rennet.
Appl. Microbiol. Biotechnol., 101:6951-6968, 2017

PubMed Abstract: The potential of using a synthetic cardosin-based rennet in cheese manufacturing was recently demonstrated with the development and optimization of production of a recombinant form of cardosin B in Kluyveromyces lactis. With the goal of providing a more detailed characterization of this rennet, we herein evaluate the impact of the plant-specific insert (PSI) on cardosin B secretion in this yeast, and provide a thorough analysis of the specificity requirements as well as the biochemical and structural properties of the isolated recombinant protease. We demonstrate that the PSI domain can be substituted by different linker sequences without substantially affecting protein secretion and milk clotting activity. However, the presence of small portions of the PSI results in dramatic reductions of secretion yields in this heterologous system. Kinetic characterization and specificity profiling results clearly suggest that synthetic cardosin B displays lower catalytic efficiency and is more sequence selective than native cardosin B. Elucidation of the structure of synthetic cardosin B confirms the canonical fold of an aspartic protease with the presence of two high mannose-type, N-linked glycan structures; however, there are some differences in the conformation of the flap region when compared to cardosin A. These subtle variations in catalytic properties and the more stringent substrate specificity of synthetic cardosin B help to explain the observed suitability of this rennet for cheese production.

PubMed: 28770303
DOI: 10.1007/s00253-017-8445-8

実験手法

X-RAY DIFFRACTION (2.375 Å)