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使用飞蝗作为新底物,米曲霉OC-7生产菊粉酶及田口设计实验法的优化过程

Inulinase production by Geotrichum candidum OC-7 using migratory locusts as a new substrate and optimization process with Taguchi DOE.

作者信息

Canli Ozden, Tasar Gani Erhan, Taskin Mesut

机构信息

Department of Biology, Ataturk University, Erzurum, Turkey.

出版信息

Toxicol Ind Health. 2013 Sep;29(8):704-10. doi: 10.1177/0748233712442737. Epub 2012 Apr 10.

DOI:10.1177/0748233712442737
PMID:22495518
Abstract

Utilization of migratory locusts (Locusta migratoria) as a main substrate due to its high protein content for inulinase (2,1-β-d-fructan fructanohydrolase) production by Geotrichum candidum OC-7 was investigated in this study. To optimize fermentation conditions, four influential factors (locust powder (LP) concentration, sucrose concentration, pH and fermentation time) at three levels were investigated using Taguchi orthogonal array (OA) design of experiment (DOE). Inulinase yield obtained from the designed experiments with regard to Taguchi L9 OA was processed with Minitab 15 software at 'larger is better' as quality character. The results showed that optimal fermentation conditions determined as LP 30 g/l, sucrose 20 g/l, pH 6.0 and time 48 h. Maximum inulinase activity was recorded as 30.12 U/ml, which was closer to the predicted value (30.56 U/ml). To verify the results, analysis of variance test was employed. LP had the greatest contribution (71.96%) among the other factors. Sucrose had lower contribution (13.96%) than LP. This result demonstrated that LP had a strong effect on inulinase activity and can be used for enzyme production. Taguchi DOE application enhanced enzyme activity to about 3.05-fold versus unoptimized condition and 2.34-fold versus control medium. Consequently, higher inulinase production can be achieved by the utilization of an edible insect material as an alternative substrate and Taguchi DOE presents suitable optimization method for biotechnological process.

摘要

本研究考察了利用蛋白质含量高的飞蝗(Locusta migratoria)作为主要底物,用于念珠地霉OC-7生产菊粉酶(2,1-β-D-呋喃果糖苷果糖水解酶)。为优化发酵条件,采用田口正交阵列(OA)实验设计(DOE)研究了三个水平的四个影响因素(蝗虫粉(LP)浓度、蔗糖浓度、pH值和发酵时间)。针对田口L9 OA设计实验获得的菊粉酶产量,使用Minitab 15软件以“越大越好”作为质量特性进行处理。结果表明,确定的最佳发酵条件为LP 30 g/l、蔗糖20 g/l、pH 6.0和时间48小时。记录的最大菊粉酶活性为30.12 U/ml,接近预测值(30.56 U/ml)。为验证结果,采用方差分析测试。LP在其他因素中贡献最大(71.96%)。蔗糖的贡献(13.96%)低于LP。该结果表明LP对菊粉酶活性有很强的影响,可用于酶的生产。与未优化条件相比,田口DOE的应用使酶活性提高了约3.05倍,与对照培养基相比提高了2.34倍。因此,利用可食用昆虫材料作为替代底物可实现更高的菊粉酶产量,田口DOE为生物技术过程提供了合适的优化方法。

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