第
11卷，第
1期
2021年
1月Vol. 11,
No. 1
Jan. ,
2021环境工程技术学报
Journal of Environmental Engineering Technology
杨凯，张硕
，崔康平
，等•活性污泥胞外聚合物中蛋白质定量方法的评价& J •环境工程技术学报
,2021
,11
(1
):144-150.
YANG K, ZHANG S, CUI K P, et al. Evaluation of protein quantification metUods for extracellular polymers in activated sludge & J'. Journal of
Environmental Engineering Technology
,2021
,11
(1
) : 144-150.
活性污泥胞外聚合物中蛋白质定量方法的评价
杨凯
1,张硕
2，崔康平
1,黑生强
2，吕学敏
2,宋广清
2,张弦
2，黄霞
2 *
收稿日期
：2020-13-09
基金项目
：科学技术部中日国际项目
(2016YFE0118500
)
作者简介:杨凯
(1993
—)，男，硕士
，研究方向为膜污染机理
,yangkai986155252@ 163. com
*责任作者:黄霞
(1965
—)，女
，教授
，博士
，研究方向为膜法水与废水处理新技术的研究与应用
,xhuang@ tsinghua. edu. cn1.合肥工业大学资源与环境工程学院
2.环境模拟与污染控制国家重点联合实验室，清华大学环境学院
摘要 胞外聚合物(
EPS)中蛋白质的准确定量对研究活性污泥性质具有重要意义。为对
EPS中蛋白质定量方法的选择提供
依据，系统评价了
Modified Lowry法、
BCA法、
Bradford法的测试性能，包括测试范围、准确度、精密度及对
EPS中部分组分
(
Cf
2+、
Mg
u、还原性糖、腐殖酸)的抗干扰性。结果表明:
Modified Lowry法在蛋白质浓度为
定量测定蛋白质含量的方法及原理
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定量测定蛋白质含量的方法及原理
在生物化学和分子生物学研究中，准确测定样品中蛋白质的含量至关重要。本文将介绍几种常用的定量测定蛋白质含量的方法及其原理。
Mercury (Hg2+) effect on enzyme activities and hepatopancreas histo structures of juvenile
Chinese mitten crab Eriocheir sinensis*
Abstract We studied the effects of mercury (Hg2+) on antioxidant and digestive enzyme
activities in terms of LC50 value and on hepatopancreas histo structures of juvenile Chinese
mitten crabs Eriocheir sinensis in 40-day exposure to various concentrations of Hg2+ (0, 0.01,
0.05, 0.10, 0.20, and 0.30 mg/L). The results show that the activities of superoxide dismutase
(SOD), glutathione peroxidase (GPX) and catalase (CAT) significantly increased in the
concentrations of 0.01 and 0.05 mg/L, while that of enzyme decreased in 0.10, 0.20 and 0.30
mg/L treatments. Meanwhile, Hg2+ disrupted the histo structures of the hepatopancreas,
causing decreases in activities of pepsin, tryptase, amylase, and cellulose, which are synthesized
Bradford法蛋白定量（Bradford Protein Assay ）
Bradford Assay is a rapid and accurate method commonly used to determine the total
protein concentration of a sample. The assay is based on the observation that the
absorbance maximum for an acidic solution of Coomassie Brilliant Blue G-250 shifts
from 465 nm to 595 nm when binding to protein occurs. Both hydrophobic and ionic
interactions stabilize the anionic form of the dye, causing a visible color change.
Within the linear range of the assay (~5-25 mcg/mL), the more protein present, the
more Coomassie binds.
Reference
Bradford, M. A rapid and sensitive method for the quantitation of microgram
quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem.
(1976) 72, 248-254.
考马斯亮蓝染色法 (Bradford法)测定蛋白质含量 原理
1976年Bradford建立了用考马斯亮蓝G250与蛋白质结合的原理，迅速、敏感的定量测定蛋白质的方法。染料与蛋白质结合后引起染料最大吸收的改变，从465nm变为595nm，光吸收增加。蛋白质-染料复合物具有高的消光系数，因此大大提高了蛋白质测定的灵敏度，最低检出量为1μg蛋白。染料与蛋白质的结合是很迅速的过程，大约需2min，结合物的颜色在1h内是稳定的。一些阳离子，如K+，Na+，Mg2+，(NH4)2SO4，乙醇等物质不干扰测定，而大量的去污剂如TritonX100，SDS等严重干扰测定，少量的去污剂可通过用适当的对照而消除。由于染色法简单迅速，干扰物质少，灵敏度高，现已广泛应用于蛋白质含量的测定。
