Santerre Henriksen A L, Carlsen M, de Bang H, Nielsen J
Center for Process Biotechnology, Department of Biotechnology, Building 223, Technical University of Denmark, DK-2800 Lyngby, Denmark.
Biotechnol Bioeng. 1999 Oct 5;65(1):76-82. doi: 10.1002/(sici)1097-0290(19991005)65:1<76::aid-bit9>3.0.co;2-d.
Pulse and pulse-chase experiments have been performed to study L-[(35)S] methionine incorporation and protein secretion kinetics in Aspergillus oryzae. Pulse experiments confirmed the mechanism of methionine uptake reported previously for Penicillium chrysogenum (Benko et al., 1967). Pulse-chase experiments were carried out to investigate the alpha-amylase secretion kinetics in A. oryzae. No unglycosylated alpha-amylase was detected neither intracellularly nor extracellularly demonstrating that glycosylation was not the rate controlling step in the secretory pathway. The pulse chase experiments indicated that there are two pools of intracellular alpha-amylase: a fast secreted and a slow secreted. The secretion of those two pools were described with a kinetic model, which was fitted to the pulse chase experiments.
已进行脉冲和脉冲追踪实验，以研究米曲霉中L-[(35)S]甲硫氨酸掺入和蛋白质分泌动力学。脉冲实验证实了先前报道的产黄青霉甲硫氨酸摄取机制（本科等人，1967年）。进行脉冲追踪实验以研究米曲霉中α-淀粉酶的分泌动力学。在细胞内和细胞外均未检测到未糖基化的α-淀粉酶，这表明糖基化不是分泌途径中的速率控制步骤。脉冲追踪实验表明，细胞内有两池α-淀粉酶：快速分泌池和缓慢分泌池。用动力学模型描述了这两池的分泌情况，该模型与脉冲追踪实验拟合。
