CICECO-Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193, Aveiro, Portugal.
Department of Engineering Bioprocess and Biotechnology, School of Pharmaceutical Sciences, UNESP - São Paulo State University, Araraquara, Brazil.
Appl Microbiol Biotechnol. 2021 Jun;105(11):4515-4534. doi: 10.1007/s00253-021-11359-y. Epub 2021 Jun 1.
In the past decades, the production of biopharmaceuticals has gained high interest due to its great sensitivity, specificity, and lower risk of negative effects to patients. Biopharmaceuticals are mostly therapeutic recombinant proteins produced through biotechnological processes. In this context, L-asparaginase (L-asparagine amidohydrolase, L-ASNase (E.C. 3.5.1.1)) is a therapeutic enzyme that has been abundantly studied by researchers due to its antineoplastic properties. As a biopharmaceutical, L-ASNase has been used in the treatment of acute lymphoblastic leukemia (ALL), acute myeloblastic leukemia (AML), and other lymphoid malignancies, in combination with other drugs. Besides its application as a biopharmaceutical, this enzyme is widely used in food processing industries as an acrylamide mitigation agent and as a biosensor for the detection of L-asparagine in physiological fluids at nano-levels. The great demand for L-ASNase is supplied by recombinant enzymes from Escherichia coli and Erwinia chrysanthemi. However, production processes are associated to low yields and proteins associated to immunogenicity problems, which leads to the search for a better enzyme source. Considering the L-ASNase pharmacological and food importance, this review provides an overview of the current biotechnological developments in L-ASNase production and biochemical characterization aiming to improve the knowledge about its production. KEY POINTS: • Microbial enzyme applications as biopharmaceutical and in food industry • Biosynthesis process: from the microorganism to bioreactor technology • Enzyme activity and kinetic properties: crucial for the final application.
在过去的几十年中，由于其高度的灵敏度、特异性和对患者产生副作用的风险较低，生物制药的生产引起了广泛关注。生物制药主要是通过生物技术过程生产的治疗性重组蛋白。在这种情况下，L-天冬酰胺酶（L-天冬酰胺酰胺水解酶，L-ASNase（E.C.3.5.1.1））是一种治疗性酶，由于其抗肿瘤特性，已被研究人员广泛研究。作为一种生物制药，L-ASNase 已与其他药物联合用于治疗急性淋巴细胞白血病（ALL）、急性髓细胞白血病（AML）和其他淋巴恶性肿瘤。除了作为生物制药的应用外，该酶还广泛用于食品加工行业，作为丙烯酰胺缓解剂和用于生理流体中 L-天冬酰胺的纳米级检测的生物传感器。对 L-ASNase 的巨大需求由大肠杆菌和菊欧文氏菌的重组酶供应。然而，生产过程与低产量和与免疫原性问题相关的蛋白质有关，这导致人们寻找更好的酶源。考虑到 L-ASNase 的药理学和食品重要性，本综述提供了 L-ASNase 生产的当前生物技术发展和生化特性的概述，旨在提高对其生产的认识。要点：•微生物酶在生物制药和食品工业中的应用•生物合成过程：从微生物到生物反应器技术•酶活性和动力学特性：对最终应用至关重要。
