从硅藻中提取和纯化核酮糖-1,5-二磷酸羧化酶/加氧酶
Rubisco Extraction and Purification from Diatoms.
作者信息
Young Jodi N, Heureux Ana M C, Rickaby Rosalind E M, Morel François M M, Whitney Spencer M, Sharwood Robert E
机构信息
Department of Oceanography, University of Washington, Seattle, USA.
Department of Earth Sciences, University of Oxford, Oxford, UK.
出版信息
Bio Protoc. 2017 Mar 20;7(6):e2191. doi: 10.21769/BioProtoc.2191.
This protocol describes a method to extract ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) from diatoms () to determine catalytic performance. This protocol has been adapted from use in cyanobacteria and higher plants (Andrews, 1988; Whitney and Sharwood, 2007). First part (steps A1-A3) of the extraction provides a crude extract of Rubisco that is sufficient for carboxylation assays to measure the Michaelis constant for CO (K) and the catalytic turnover rate ( ). However, the further purification steps outlined (steps B1-B4) are needed for measurements of Rubisco CO/O Specificity (S, [ Kane , 1994 ]).
本方案描述了一种从硅藻中提取1,5 - 二磷酸核酮糖羧化酶加氧酶（Rubisco）以测定其催化性能的方法。该方案改编自用于蓝细菌和高等植物的方法（安德鲁斯，1988年；惠特尼和沙伍德，2007年）。提取的第一部分（步骤A1 - A3）提供了足够用于羧化测定以测量CO的米氏常数（K）和催化周转速率（ ）的Rubisco粗提物。然而，对于Rubisco CO/O特异性（S，[凯恩，1994年]）的测量，需要概述的进一步纯化步骤（步骤B1 - B4）。
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