Martins das Neves Luiz Carlos, Pessoa Adalberto, Vitolo Michele
Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical Sciences, University of São Paulo, Av. Prof. Lineu Prestes, 580, B-16, 05508-900 São Paulo, SP, Brazil.
Biotechnol Prog. 2005 Jul-Aug;21(4):1136-9. doi: 10.1021/bp050034g.
In a 5-L fermentor (NBS-MF 105), Saccharomyces cerevisiae W303-181 (1.0 g dry matter/L) was inoculated into 3.0 L of liquid medium containing glucose (10 or 20 g/L), yeast nitrogen base (YNB, 3.7 or 7.4 g/L), l-histidine (0.02 g/L), l-tryptophan (0.02 g/L), uracil (0.02 g/L), and adenine (0.02 g/L). The culture was carried out batchwise for 12 or 24 h at 30 degrees C, pH 4.6 or 5.7, aeration of 0, 0.8, 1.7 or 2.2 vvm, and agitation of 400 rpm. The highest G6PDH productivity (10.5 U/L.h) and specific activity (320 U/mg of protein) occurred at aeration of 2.2 vvm, pH 5.7, 10 g/L of glucose, and 3.7 g/L of YNB. The G6PDH specific activity attained was comparable with those of commercial preparations, which are between 50 and 600 U/mg of protein.
在一个5升的发酵罐(NBS-MF 105)中,将酿酒酵母W303-181(1.0克干物质/升)接种到3.0升含有葡萄糖(10或20克/升)、酵母氮源(YNB,3.7或7.4克/升)、L-组氨酸(0.02克/升)、L-色氨酸(0.02克/升)、尿嘧啶(0.02克/升)和腺嘌呤(0.02克/升)的液体培养基中。在30℃、pH值4.6或5.7、通气量为0、0.8、1.7或2.2体积/体积·分钟、搅拌速度为400转/分钟的条件下分批培养12或24小时。在通气量为2.2体积/体积·分钟、pH值5.7、葡萄糖浓度为10克/升、YNB浓度为3.7克/升时,葡萄糖-6-磷酸脱氢酶(G6PDH)的最高生产率(10.5单位/升·小时)和比活性(320单位/毫克蛋白质)出现。所获得的G6PDH比活性与商业制剂相当,商业制剂的比活性在50至600单位/毫克蛋白质之间。