在大豆农工业残渣中扩大异形曲霉 URM0269 胶原酶的优化生产规模。

IF 7.7 1区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY International Journal of Biological Macromolecules Pub Date : 2024-11-16 DOI:10.1016/j.ijbiomac.2024.137734
Lígia Maria Gonçalves Fernandes, Jônatas de Carvalho-Silva, William Eugenio Lopes da Silva, Márcia Nieves Carneiro da Cunha, Attilio Converti, Tatiana Souza Porto
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引用次数: 0

摘要

以黄豆粉为底物,通过浸没式发酵优化了异形曲霉 URM0269 产出的胶原酶和蛋白酶。以底物和酵母提取物的浓度为自变量,按照复合设计进行发酵。在搅拌罐生物反应器中放大最佳条件,以评估发酵动力学。当底物浓度为 2.0 %、酵母提取物浓度为 0.1 % 时,两种酶的产量最高。相反,规模扩大后,胶原蛋白酶活性从 33.5 U/mL增至 148.5 U/mL,而蛋白酶活性则从 16.3 U/mL降至 11.7 U/mL。异形菌 URM0269 的最大生长速率为 0.09 h-1,65 h 后生物量上的蛋白酶和胶原酶产量分别为 2.70 U/mgx和 34.22 U/mgx。胶原蛋白酶以胶原蛋白为底物,在温度为 40 ℃、pH 为 6.0 的条件下发挥最佳作用,并呈现出异构趋势,最大反应速率为 132.47 U/mL。测定了最佳温度下胶原蛋白降解的热力学参数,如吉布斯自由能(74.16 kJ/mol)、焓(11.64 kJ/mol)、熵(-199.63 J/K.mol)和活化能(14.25 kJ/mol)。这些结果表明,大豆粉是生产胶原蛋白酶的潜在农工业残留物。此外,这种胶原蛋白酶在生物化学和热力学方面的特点也很有希望用于其他生物技术领域。
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Scaling up the optimized production of Aspergillus heteromorphus URM0269 collagenase in soybean agroindustrial residue.

Collagenase and protease productions from Aspergillus heteromorphus URM0269 were optimized by submerged fermentation using soybean flour as substrate. Fermentations were performed according to composite design using the concentrations of substrate and yeast extract as the independent variables. The best condition was scaled up in a stirred tank bioreactor to assess the fermentation kinetics. The highest production of both enzymes occurred at concentrations of 2.0 % substrate and 0.1 % yeast extract. Contrariwise, after scale-up, collagenase activity increased from 33.5 to 148.5 U/mL, while the protease decreased from 16.3 to 11.7 U/mL. A. heteromorphus URM0269 showed a maximum growth rate of 0.09 h-1 and yields of protease and collagenase on biomass, after 65 h of 2.70 and 34.22 U/mgx, respectively. Collagenase acted optimally at 40 °C and pH 6.0 on collagen as a substrate and displayed an allosteric trend, achieving a maximum reaction rate of 132.47 U/mL. Thermodynamic parameters of collagen degradation such as Gibbs free energy (74.16 kJ/mol), enthalpy (11.64 kJ/mol), entropy (-199.63 J/K.mol), and activation energy (14.25 kJ/mol) were determined for optimal temperature. These results demonstrated that soybean flour is a potential agroindustrial residue for collagenase production. Furthermore, the collagenase displayed promising biochemical and thermodynamic features for other biotechnological applications.

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来源期刊
International Journal of Biological Macromolecules
International Journal of Biological Macromolecules 生物-生化与分子生物学
CiteScore
13.70
自引率
9.80%
发文量
2728
审稿时长
64 days
期刊介绍: The International Journal of Biological Macromolecules is a well-established international journal dedicated to research on the chemical and biological aspects of natural macromolecules. Focusing on proteins, macromolecular carbohydrates, glycoproteins, proteoglycans, lignins, biological poly-acids, and nucleic acids, the journal presents the latest findings in molecular structure, properties, biological activities, interactions, modifications, and functional properties. Papers must offer new and novel insights, encompassing related model systems, structural conformational studies, theoretical developments, and analytical techniques. Each paper is required to primarily focus on at least one named biological macromolecule, reflected in the title, abstract, and text.
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