开发基于具有酰胺酶活性的固定化细菌细胞的生物选择剂,用于生物检测丙烯酰胺

IF 1 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Applied Biochemistry and Microbiology Pub Date : 2024-09-22 DOI:10.1134/S0003683824605031
E. M. Protasova, Yu. G. Maksimova
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引用次数: 0

摘要

通过在凝胶结构中夹带海藻酸钡和琼脂糖,以及在热膨胀石墨(TEG)上形成生物膜,固定了具有酰胺酶活性的放线菌细胞红球菌 4-1 和红球菌 11-2 以及蛋白细菌粪绿球菌 2。测定了这种固定化生物催化剂在冷冻和脱水储存后的操作稳定性,并开发了基于这种生物选择剂的电导丙烯酰胺生物传感器原型。最理想的固定化细胞储存方法是在 -20 至 -80°C 的温度下冷冻;也可在 4-25°C 的温度下湿润长期储存。结果表明,这些细胞最适合用于固定在琼脂糖凝胶结构中的丙烯酰胺粪肠球菌 2 的生物检测。与海藻酸钡凝胶相比,固定了细菌细胞的琼脂糖凝胶在丙烯酰胺转化为丙烯酸的连续循环过程中具有更高的机械强度和稳定性。在细胞固定过程中添加碳纳米材料可以增强海藻酸钡凝胶的机械强度。在用于制造电极的碳材料上生长生物膜也很有前景。红喉杆菌 11-2 的生物膜在 TEG 上能够在 20 多个反应循环中将丙烯酰胺转化为丙烯酸,同时保持至少 50% 的酰胺酶活性。
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Development of a Bio-Selecting Agent Based on Immobilized Bacterial Cells with Amidase Activity for Bio-Detection of Acrylamide

Actinobacteria cells Rhodococcus erythropolis 4-1 and Rhodococcus erythropolis 11-2 and Proteobacteria Alcaligenes faecalis 2, which have amidase activity, were immobilized by entrapping barium alginate and agarose into the gel structure, as well as by obtaining biofilms on thermally expanded graphite (TEG). The operational stability of such immobilized biocatalysts after storage in frozen and dehydrated form was determined, and a prototype of a conductometric acrylamide biosensor based on such a bioselective agent was developed. The most preferred method for storing immobilized cells was freezing at temperatures from –20 to –80°C; long-term storage is also possible wet at 4–25°C. It was shown that these cells were most preferable for the biodetection of acrylamide A. faecalis 2, immobilized in an agarose gel structure. An agarose gel with bacterial cells immobilized in its structure had greater mechanical strength and stability during successive cycles of conversion of acrylamide into acrylic acid compared to barium alginate gel. The mechanical strength of barium alginate gel can be enhanced by the addition of carbon nanomaterials during cell immobilization. Growing biofilms on carbon materials used for manufacturing electrodes is also promising. Biofilms of R. erythropolis 11-2 on TEG are capable of converting acrylamide into acrylic acid in more than 20 reaction cycles while maintaining at least 50% amidase activity.

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来源期刊
Applied Biochemistry and Microbiology
Applied Biochemistry and Microbiology 生物-生物工程与应用微生物
CiteScore
1.70
自引率
12.50%
发文量
75
审稿时长
6-12 weeks
期刊介绍: Applied Biochemistry and Microbiology is an international peer reviewed journal that publishes original articles on biochemistry and microbiology that have or may have practical applications. The studies include: enzymes and mechanisms of enzymatic reactions, biosynthesis of low and high molecular physiologically active compounds; the studies of their structure and properties; biogenesis and pathways of their regulation; metabolism of producers of biologically active compounds, biocatalysis in organic synthesis, applied genetics of microorganisms, applied enzymology; protein and metabolic engineering, biochemical bases of phytoimmunity, applied aspects of biochemical and immunochemical analysis; biodegradation of xenobiotics; biosensors; biomedical research (without clinical studies). Along with experimental works, the journal publishes descriptions of novel research techniques and reviews on selected topics.
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