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从巴西卡廷加生物群落中分离出的土曲霉 S-18 菌株发酵生产 l-天冬酰胺酶的筛选与优化。

Screening and optimizing fermentation production of l-asparaginase by Aspergillus terreus strain S-18 isolated from the Brazilian Caatinga Biome.

机构信息

State University of Paraíba, Campina Grande, PB, Brazil.

Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, Brazil.

出版信息

J Appl Microbiol. 2019 May;126(5):1426-1437. doi: 10.1111/jam.14221. Epub 2019 Mar 19.

Abstract

AIMS

The aim of this study was to find new eukaryotic sources of the l-asparaginase (l-ASNase), since the prokaryotic sources of the enzyme are well-reported as causing allergic hypersensitivity reactions in a significant number of patients. This report describes screening for l-ASNase production by filamentous fungi isolated from the Brazilian Caatinga, and the optimization of fermentation parameters to increase fungal growth and improve yield in the production of l-ASNase.

METHODS AND RESULTS

Thirty-two filamentous fungi were investigated in this study. When Aspergillus terreus strain S-18 was cultured in a proline-enriched medium, intracellular l-ASNase was expressed in concurrence with reduced l-glutaminase (l-GLUase) and protease activities. Fermentation conditions were then optimized in a 5-l bioreactor system to produce a maximum volumetric yield of 108 U total of l-ASNase activity.

CONCLUSIONS

The work reported here represents the first attempt to produce l-ASNase by filamentous fungi isolated from Brazil and offers a promising alternative eukaryotic source for l-ASNase production.

SIGNIFICANCE AND IMPACT OF THE STUDY

In order to minimize the side effects caused by bacterial l-ASNase, the search of eukaryotic micro-organism for l-ASNase was carried out in fungi. This study demonstrates the diversity of filamentous fungi isolated from the Brazilian Caatinga Biome and the importance of knowledge of the microbial metabolism to obtain high concentrations of biotechnological products.

摘要

目的

本研究旨在寻找新的真核来源的 l-天冬酰胺酶(l-ASNase),因为该酶的原核来源已被充分报道,会导致相当数量的患者发生过敏超敏反应。本报告描述了从巴西卡廷加分离的丝状真菌中筛选 l-ASNase 产生的情况,并优化了发酵参数,以增加真菌生长并提高 l-ASNase 生产中的产量。

方法和结果

本研究共研究了 32 株丝状真菌。当 A. terreus 菌株 S-18 在富含脯氨酸的培养基中培养时,细胞内 l-ASNase 与降低的 l-谷氨酰胺酶(l-GLUase)和蛋白酶活性同时表达。然后在 5 升生物反应器系统中优化发酵条件,以产生最大的 108 U 总 l-ASNase 活性的比容产率。

结论

本报告首次尝试使用从巴西分离的丝状真菌生产 l-ASNase,并为生产 l-ASNase 提供了一种有前途的真核来源。

研究的意义和影响

为了最大限度地减少细菌 l-ASNase 引起的副作用,对真菌中的 l-ASNase 进行了真核微生物的搜索。本研究证明了从巴西卡廷加生物群落中分离出的丝状真菌的多样性,以及了解微生物代谢以获得高浓度生物技术产品的重要性。

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