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基于响应表面方法,优化了从Fusarium胎儿中L-asparaginase的生产,净化和表征

Javaraiah Parashiva¹, Bettadapura Rameshgowda Nuthan¹, Madeva Bharatha²

¹Department of Studies in Microbiology, University of Mysore, Manasagangotri, Mysuru, Karnataka, 570 006, India.

World journal of microbiology & biotechnology

|July 13, 2023

概括

这项研究优化了Fusarium胎儿的L-asparaginase生产,使其活性增加了2.65倍. 纯化的酶具有很高的特异性,并且在癌症治疗中具有大规模生物处理的潜力.

科学领域:

生物化学生物化学
酶学是一种酶学.
生物技术是生物技术.

背景情况:

L-阿斯巴拉金酶是急性淋巴细胞白血病的关键化疗剂.
细菌L-asparaginase可以引起过敏反应和副作用.
目前正在探索L-asparaginase的替代来源,以改善治疗结果.

研究的目的:

为了优化生长参数,增强Fusarium胎儿的L-阿斯巴拉基纳酶生产.
为了净化和表征从Fusarium foetens中的L-asparaginase.
评估Fusarium胎儿在大规模L-asparaginase生物处理中的潜力.

主要方法:

优化L-asparaginase的生产使用一个因素在一个时间和响应表面的方法 (中央复合设计).
使用DEAE-纤维素列染色学净化L-阿斯巴拉金酶.
纯化L-阿斯巴拉金酶的表征,包括分子质量测定 (SDS-PAGE),最佳pH和温度,基质特异性和动力参数 (Km,Vmax).

主要成果:

在27.5°C,pH8,1.5×10⁶子/ml注射液下达到12.83 IU/ml的最大L-阿斯巴拉金酶活性,并进行了7天的潜伏.
优化导致L-asparaginase活性增加了2.65倍,与未优化的条件相比.
纯化L-氨酸酶 (15.60倍) 的活性为127.26 IU/ml,分子质量为~37 kDa,在pH 5和40°C时的最佳活性,以及对L-氨酸的高特异性.

结论:

类胎儿是生产高活性L-asparaginase的有希望的来源.
优化的生产和净化方法显著提高了酶产量和特异性活性.
来自Fusarium foetens的特征L-asparaginase表明了用于治疗应用的具有成本效益的大规模生物处理的潜力.

关键词:

专色谱学专色谱学专色谱学酶活性的酶活动.动力常数动力常数在L-asparaginase的使用中.响应表面的方法响应表面方法

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