Preliminary Study on Polymerization between Hemoglobin and Enzymes during the Preparation of PolyHb-SOD-CAT-CA

IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Doklady Biochemistry and Biophysics Pub Date : 2024-08-28 DOI:10.1134/S1607672924600477
Lili Zhang, Renci Tian, Jiawei Xiao, Yaoxi Wang, Kun Feng, Gang Chen
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Abstract

The objective of this study was to explore the influence of different factors on the aggregation effect on hemoglobin (Hb) and enzymes during the preparation of Polyhemoglobin-Superoxide dismutase-Catalase-Carbonic anhydrase (PolyHb-SOD-CAT-CA). Several factors including temperatures, pH values, Glutaraldehyde (GDA) amounts and enzymes amounts were investigated systematically to study their effects on the enzymes recoveries and polymerization rates including the Superoxide dismutase (SOD), Catalase (CAT) and Carbonic anhydrase (CA), as well as their effects on the molecular weight distribution of PolyHb-SOD-CAT-CA. Then the oxygen affinity and methemoglobin (MetHb) contents of obtained PolyHb-SOD-CAT-CA were measured to evaluate the effects of enzyme crosslinking on the properties of Polyhemoglobin (PolyHb) moieties in the molecular structure of obtained PolyHb-SOD-CAT-CA conjugate. The results showed that the enzyme recoveries and polymerization rates could be decreased with the temperatures increasing and could be generally kept stable in the physiological pH conditions, but presented only slight changes among the investigated enzyme amounts ranges. Although the GDA concentration increasing could promote the enzyme polymerization rates, the enzyme recoveries decreased in whole. The polymerization rate and molecular size of PolyHb-SOD-CAT-CA conjugate increased with the elevation of temperature and the concentration of GDA. Lastly, the P50 values, Hill coefficients, and MetHb contents of PolyHb-SOD-CAT-CA conjugate with different enzyme crosslinking degrees exhibited no obvious differences with each other. In conclusion, the polymerization reactions between enzymes and Hb molecules could be remarkably affected by temperatures, pH values, and GDA amounts, and the enzyme crosslinking presented no obvious effects on the Hb properties, especially about the oxygen affinity and oxidation degrees.

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制备 PolyHb-SOD-CAT-CA 过程中血红蛋白与酶之间聚合作用的初步研究
本研究旨在探讨在制备聚血红蛋白-超氧化物歧化酶-催化酶-碳酸酐酶(PolyHb-SOD-CAT-CA)的过程中,不同因素对血红蛋白(Hb)和酶的聚集效应的影响。研究人员系统地考察了温度、pH 值、戊二醛(GDA)用量和酶用量等因素对超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和碳酸酐酶(CA)等酶的回收率和聚合率的影响,以及它们对聚合血红蛋白-超氧化物歧化酶-过氧化氢酶-碳酸酐酶(PolyHb-SOD-CAT-CA)分子量分布的影响。然后测定了所得 PolyHb-SOD-CAT-CA 的氧亲和力和高铁血红蛋白(MetHb)含量,以评估酶交联对所得 PolyHb-SOD-CAT-CA 共轭物分子结构中高铁血红蛋白(PolyHb)分子性能的影响。结果表明,酶的回收率和聚合率会随着温度的升高而降低,在生理 pH 条件下基本保持稳定,但在所研究的酶用量范围内变化不大。虽然 GDA 浓度的增加会促进酶的聚合速率,但酶的回收率却整体下降。随着温度和 GDA 浓度的升高,PolyHb-SOD-CAT-CA 共轭物的聚合速率和分子大小都有所增加。最后,不同酶交联度的 PolyHb-SOD-CAT-CA 共轭物的 P50 值、希尔系数和 MetHb 含量均无明显差异。总之,酶与 Hb 分子的聚合反应会受到温度、pH 值和 GDA 含量的显著影响,而酶交联对 Hb 性能,尤其是氧亲和性和氧化度没有明显影响。
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来源期刊
Doklady Biochemistry and Biophysics
Doklady Biochemistry and Biophysics 生物-生化与分子生物学
CiteScore
1.60
自引率
12.50%
发文量
68
审稿时长
6-12 weeks
期刊介绍: Doklady Biochemistry and Biophysics is a journal consisting of English translations of articles published in Russian in biochemistry and biophysics sections of the Russian-language journal Doklady Akademii Nauk. The journal''s goal is to publish the most significant new research in biochemistry and biophysics carried out in Russia today or in collaboration with Russian authors. The journal accepts only articles in the Russian language that are submitted or recommended by acting Russian or foreign members of the Russian Academy of Sciences. The journal does not accept direct submissions in English.
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