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毛霉属和青霉属物种的磷酸果糖激酶与葡萄糖分解代谢

Phosphofrucktokinase and glucose catabolism of Mucor and Penicillium species.

作者信息

Boonsaeng V, Sullivan P A, Shepherd M G

出版信息

Can J Microbiol. 1977 Sep;23(9):1214-24. doi: 10.1139/m77-182.

Abstract

The primary catabolic pathways in the fungi Penicillium notatum and P. duponti, and Mucor rouxii and M. miehei were examined by measuring the relative rate of 14CO2 production from different carbon atoms of specifically labelled glucose. It was found that these organisms dissimilate glucose predominantly via the Embden--Meyerhof pathway in conjunction with the tricarboxylic acid cycle and to a lesser extent by the pentose phosphate pathway. Phosphofructokinase (EC 2.7.1.11) activity could not be detected initially in Penicillium species because of the interference from mannitol-1-phosphate dehydrogenase (EC 1.1.1.17) and NADH oxidase (EC 1.6.99.3). A combination of differential centrifuging and a heat treatment of Penicillium cell-free extracts in the presence of fructose-6-phosphate removed the interfering enzymes. The kinetic characteristics of phosphofructokinase from P. notatum and M. rouxii are described. The enzyme presents highly cooperative kinetics for fructose-6-phosphate. The kinetics for ATP show no cooperativity and inhibition by excess ATP is observed. The addition of AMP activated the P. notatum enzyme, relieving ATP inhibition; slight inhibition by AMP was observed with the M. rouxii enzyme. In contrast M. rouxii pyruvate kinase (EC 2.7.1.40) is activated 50-fold by fructose-1,6-diphosphate whereas pyruvate kinase from P. notatum and P. duponti were unaffected by fructose-1,6-diphosphate.

摘要

通过测量特定标记葡萄糖不同碳原子产生(^{14}CO_2)的相对速率,研究了青霉属的点青霉和杜邦青霉以及鲁氏毛霉和米黑毛霉中的主要分解代谢途径。结果发现,这些生物体主要通过糖酵解途径与三羧酸循环相结合来异化葡萄糖,较少程度上通过磷酸戊糖途径。由于1-磷酸甘露醇脱氢酶(EC 1.1.1.17)和NADH氧化酶(EC 1.6.99.3)的干扰,最初在青霉属物种中未检测到磷酸果糖激酶(EC 2.7.1.11)的活性。在存在6-磷酸果糖的情况下,对青霉无细胞提取物进行差速离心和热处理相结合的方法去除了干扰酶。描述了点青霉和鲁氏毛霉中磷酸果糖激酶的动力学特性。该酶对6-磷酸果糖呈现高度协同动力学。ATP的动力学没有协同性,并且观察到过量ATP的抑制作用。添加AMP激活了点青霉的酶,解除了ATP的抑制作用;对于鲁氏毛霉的酶,观察到AMP有轻微抑制作用。相比之下,鲁氏毛霉的丙酮酸激酶(EC 2.7.1.40)被1,6-二磷酸果糖激活了50倍,而点青霉和杜邦青霉的丙酮酸激酶不受1,6-二磷酸果糖的影响。

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