M(III)促进中温及嗜热生物来源酶的回收与浓缩。

M(III)-facilitated recovery and concentration of enzymes from mesophilic and thermophilic organisms.

作者信息

Collingwood T N, Shanmugam M, Daniel R M, Langdon A G

机构信息

Chemistry Department, University of Waikato, Hamilton, New Zealand.

出版信息

J Biochem Biophys Methods. 1989 Oct;19(4):281-6. doi: 10.1016/0165-022x(89)90059-6.

DOI:10.1016/0165-022x(89)90059-6

PMID:2515216

Abstract

Six enzymes isolated from organisms of widely differing thermal growth optima were flocculated from solution at constant pH by addition of Fe(III) solution. In all cases the enzyme concentration was 1 g.l-1 or less. Flocculation profiles were generated for each enzyme over a range of Fe(III) levels. The concentrated enzymes were recovered from the Fe(III)/protein complex by solubilisation with citrate and dithionite followed by precipitation with ammonium sulphate. In all cases approximately 70-80% enzyme recovery was achieved. Enzyme thermal stability did not appear to be important and protein concentration had no effect on the efficiency of enzyme recovery over the range of 0.01-1 g.l-1. Approximately 30 mmol Fe(III)/l of enzyme solution facilitated optimal enzyme recovery for all solutions studied. For protein concentrations up to 1 g.l-1 a 100-fold enzyme concentration factor can be expected.

摘要

从具有广泛不同最佳热生长温度的生物体中分离出的六种酶，通过添加铁（III）溶液在恒定pH值下从溶液中絮凝出来。在所有情况下，酶浓度均为1 g·l⁻¹或更低。针对每种酶在一系列铁（III）水平下生成了絮凝曲线。通过用柠檬酸盐和连二亚硫酸盐溶解，然后用硫酸铵沉淀，从铁（III）/蛋白质复合物中回收浓缩的酶。在所有情况下，酶回收率约为70 - 80%。酶的热稳定性似乎并不重要，并且在0.01 - 1 g·l⁻¹范围内，蛋白质浓度对酶回收效率没有影响。对于所研究的所有溶液，约30 mmol铁（III）/l的酶溶液有助于实现最佳酶回收。对于高达1 g·l⁻¹的蛋白质浓度，预计酶浓度因子可达100倍。