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添加尿嘧啶对土壤杆菌蛋白质组图谱及1,3-β-葡聚糖产生的影响

Effect of uracil addition on proteomic profiles and 1,3-β-glucan production in Agrobacterium sp.

作者信息

Jin Li-Hua, Lee Jung-Heon

机构信息

College of Biotechnology, Beijing Polytechnic, Beijing, China; Department of Chemical Engineering, Chosun University, Gwangju, Korea.

出版信息

Biotechnol Appl Biochem. 2014 May-Jun;61(3):280-8. doi: 10.1002/bab.1149. Epub 2014 Mar 25.

Abstract

Uridine diphosphate (UDP)-glucose is a precursor of 1,3-β-glucan and is synthesized from glucose-1-phosphate and uridine triphosphate (UTP). Uracil was used as a precursor for UTP, which resulted in an increase in both the UDP-glucose level and the rate of 1,3-β-glucan synthesis. 1,3-β-Glucan production metabolism was reactivated after uracil addition during fermentation. 2D-PAGE was used to examine the changes in the expression level of the key metabolic enzymes in the production of 1,3-β-glucan after uracil addition. The results showed that the expression levels of UTP-glucose-1-phosphate uridylytransferase, phosphoglucomutase, and 1,3-β-glucan synthase catalytic subunit, key metabolic enzymes in the curdlan biosynthesis pathway, were increased after uracil addition by 3.5%, 30%, and 35%, respectively. Uracil phosphoribosyltransferase, which converts uracil to uridine monophosphate (UMP), was also upregulated 79% more than that without uracil addition. However, the expression levels of orotidine 5-phosphate decarboxylase, glucose-1-phosphate adenylyltransferase, and glucose-6-phosphate isomerase were decreased after uracil addition. This proteomic information is useful for predicting changes in the pathway of uracil utilization. This work provides proteomic information for the integrative analysis of bioinformatic databases, which can be used to predict and understand the metabolism of glucan synthesis at the cellular level.

摘要

尿苷二磷酸(UDP)-葡萄糖是1,3-β-葡聚糖的前体,由葡萄糖-1-磷酸和尿苷三磷酸(UTP)合成。尿嘧啶用作UTP的前体,这导致UDP-葡萄糖水平和1,3-β-葡聚糖合成速率均增加。在发酵过程中添加尿嘧啶后,1,3-β-葡聚糖的生产代谢被重新激活。二维聚丙烯酰胺凝胶电泳(2D-PAGE)用于检测添加尿嘧啶后1,3-β-葡聚糖生产过程中关键代谢酶表达水平的变化。结果表明,添加尿嘧啶后,凝胶多糖生物合成途径中的关键代谢酶UTP-葡萄糖-1-磷酸尿苷酰转移酶、磷酸葡萄糖变位酶和1,3-β-葡聚糖合酶催化亚基的表达水平分别提高了3.5%、30%和35%。将尿嘧啶转化为尿苷单磷酸(UMP)的尿嘧啶磷酸核糖基转移酶的表达水平也比未添加尿嘧啶时上调了79%。然而,添加尿嘧啶后,乳清酸核苷5'-磷酸脱羧酶、葡萄糖-1-磷酸腺苷酰转移酶和葡萄糖-6-磷酸异构酶的表达水平降低。这些蛋白质组学信息有助于预测尿嘧啶利用途径的变化。这项工作为生物信息数据库的综合分析提供了蛋白质组学信息,可用于在细胞水平预测和理解葡聚糖合成的代谢。

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