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通过分批发酵工艺培养的转基因酿酒酵母生产葡萄糖-6-磷酸脱氢酶。

Production of glucose 6-phosphate dehydrogenase from genetically modified Saccharomyces cerevisiae grown by batch fermentation process.

作者信息

Martins das Neves Luiz Carlos, Pessoa Adalberto, Vitolo Michele

机构信息

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.

出版信息

Biotechnol Prog. 2005 Jul-Aug;21(4):1136-9. doi: 10.1021/bp050034g.

Abstract

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.

摘要

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。

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