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首页> 外文期刊>Bioprocess and Biosystems Engineering >Efficient immobilization of AGE and NAL enzymes onto functional amino resin as recyclable and high-performance biocatalyst
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Efficient immobilization of AGE and NAL enzymes onto functional amino resin as recyclable and high-performance biocatalyst

机译：将AGE和NAL酶有效地固定在功能性氨基树脂上，作为可回收的高性能生物催化剂

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摘要

N-Acetylglucosamine-2-epimerase (AGE) and N-acetylneuraminic acid lyase (NAL) were immobilized for synthesis of N-acetylneuraminic acid (Neu5Ac) on three resins: Amberzyme oxirane resin (AOR), poly (styrene-co-DVB)-Br resin (PBR) and amino resin (AR). The loading capacity and immobilized enzyme activity showed that AR was the best carrier. Three methods of glutaraldehyde cross-linking were tested and simultaneous cross-linking and immobilization was demonstrated to be the best method. The functional properties of immobilized AGE and NAL were studied and compared to those of the free enzyme. The highest enzyme activities of free and immobilized AGE were obtained in 0.1 M potassium phosphate buffer at pH 7.5 and a temperature of 37 A degrees C. Comparatively, the highest NAL activities were at pH 8.5. Meanwhile, an increase in K (m) (from 1.14 to 1.31 mg center dot mL(-1) for AGE and from 1.05 to 1.25 mg center dot mL(-1) for NAL) and a decrease in V (max) (from 177.53 to 106.37 A mu g center dot min(-1) mL(-1) for AGE and from 126.41 to 95.96 A mu g center dot min(-1) mL(-1) for NAL) were recorded after immobilization. The AR-glutaraldehyde-enzyme system exhibited better thermal stability than the free enzyme, and retained 72% of its initial activity even after eight repeated runs. The apparent activation energy (E (a)) of the free and immobilized AGE (NAL) was 117.14 kJ center dot mol(-1) (124.21 kJ center dot mol(-1)) and 78.45 kJ center dot mol(-1) (66.64 kJ center dot mol(-1)), respectively, implying that the catalytic efficiency of the immobilized enzyme was restricted by mass-transfer rather than kinetic limit. Subsequently, Neu5Ac production from GlcNAc using immobilized enzymes in one reactor was carried out resulting 101.45 g center dot L-1 of Neu5Ac and the highest conversion ratio of 82%. This method of enzyme immobilization may have a promising future for Neu5Ac production in industry.
机译：N-乙酰氨基葡萄糖-2-表异构酶（AGE）和N-乙酰神经氨酸裂解酶（NAL）被固定在三种树脂上合成N-乙酰神经氨酸（Neu5Ac）：琥珀酶环氧乙烷树脂（AOR），聚（苯乙烯-co-DVB） -溴树脂（PBR）和氨基树脂（AR）。负载能力和固定化酶活性表明，AR是最佳载体。测试了戊二醛交联的三种方法，证明同时交联和固定化是最好的方法。研究了固定化AGE和NAL的功能特性，并将其与游离酶的功能特性进行了比较。在pH 7.5和温度37 A的0.1 M磷酸钾缓冲液中获得了游离的和固定的AGE最高的酶活性。相比之下，NAL的最高活性在pH 8.5时。同时，K（m）增加（AGE的从1.14到1.31 mg中心点mL（-1），NAL的从1.05到1.25 mg中心点mL（-1））和V（max）减小（从固定后记录AGE的177.53至106.37 A中心点min（-1）mL（-1）和NAL的126.41至95.96 A中心点min（-1）mL（-1））。 AR-戊二醛-酶系统显示出比游离酶更好的热稳定性，并且即使在八次重复运行后仍保留了其初始活性的72％。游离和固定化AGE（NAL）的表观活化能（E（a））为117.14 kJ中心点mol（-1）（124.21 kJ中心点mol（-1））和78.45 kJ中心点mol（-1） （66.64 kJ中心点mol（-1）），这表明固定化酶的催化效率受传质而不是动力学极限的限制。随后，在一个反应​​器中使用固定化酶从GlcNAc生产Neu5Ac，得到101.45 g Neu5Ac的中心点L-1，最高转化率为82％。这种酶的固定化方法对于工业化生产Neu5Ac可能具有广阔的前景。

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  • 来源
    《Bioprocess and Biosystems Engineering》 |2017年第3期|331-340|共10页
  • 作者

    Cheng Jian; Zhuang Wei; Tang Chenglun; Chen Yong; Wu Jinglan; Guo Ting; Ying Hanjie;

  • 作者单位

    Nanjing Tech Univ, State Key Lab Mat Oriented Chem Engn, 5 Xinmofan Rd, Nanjing 210009, Jiangsu, Peoples R China|Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, Natl Engn Tech Res Ctr Biotechnol, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China|Nanjing Tech Univ, Synerget Innovat Ctr Adv Mat, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China;

    Nanjing Tech Univ, State Key Lab Mat Oriented Chem Engn, 5 Xinmofan Rd, Nanjing 210009, Jiangsu, Peoples R China|Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, Natl Engn Tech Res Ctr Biotechnol, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China|Nanjing Tech Univ, Synerget Innovat Ctr Adv Mat, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China;

    Nanjing Tech Univ, State Key Lab Mat Oriented Chem Engn, 5 Xinmofan Rd, Nanjing 210009, Jiangsu, Peoples R China|Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, Natl Engn Tech Res Ctr Biotechnol, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China|Nanjing Tech Univ, Synerget Innovat Ctr Adv Mat, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China;

    Nanjing Tech Univ, State Key Lab Mat Oriented Chem Engn, 5 Xinmofan Rd, Nanjing 210009, Jiangsu, Peoples R China|Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, Natl Engn Tech Res Ctr Biotechnol, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China|Nanjing Tech Univ, Synerget Innovat Ctr Adv Mat, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China;

    Nanjing Tech Univ, State Key Lab Mat Oriented Chem Engn, 5 Xinmofan Rd, Nanjing 210009, Jiangsu, Peoples R China|Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, Natl Engn Tech Res Ctr Biotechnol, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China;

    Nanjing Tech Univ, State Key Lab Mat Oriented Chem Engn, 5 Xinmofan Rd, Nanjing 210009, Jiangsu, Peoples R China|Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, Natl Engn Tech Res Ctr Biotechnol, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China;

    Nanjing Tech Univ, State Key Lab Mat Oriented Chem Engn, 5 Xinmofan Rd, Nanjing 210009, Jiangsu, Peoples R China|Nanjing Tech Univ, Coll Biotechnol & Pharmaceut Engn, Natl Engn Tech Res Ctr Biotechnol, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China|Nanjing Tech Univ, Synerget Innovat Ctr Adv Mat, 30 Puzhu South Rd, Nanjing 211816, Jiangsu, Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    N-Acetylneuraminic acid; N-Acetylneuraminic acid lyase; N-Acetylglucosamine 2-epimerase; Immobilization; Amino resin;

    机译：N-乙酰神经氨酸;N-乙酰神经氨酸裂解酶;N-乙酰氨基葡萄糖2-表异构酶;固定化;氨基树脂;

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