Metabolic engineering of Pichia pastoris X-33 for lycopene production
Metabolic engineering of Pichia pastoris X-33 for lycopene production
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DOI：
10.1016/j.procbio.2009.05.012
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发表时间：
2009-10-01
影响因子：
4.4
通讯作者：
Lee, Pyung Cheon
中科院分区：
文献类型：
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作者：
Bhataya, Anuj;Schmidt-Dannert, Claudia;Lee, Pyung Cheon
作为替代类胡萝卜素生产者，选择非胡萝卜素产生的巴斯德毕赤酵母用于红色类胡萝卜素番茄红素生产，因为它可以生长到高细胞密度而不积累乙醇，并利用各种类型的有机材料如甲醇作为碳源。设计并构建了两个合成的溶菌酶途径质粒pGAPZB-EBI* 和pGAPZB-EpBpl*p。pGAPZB-EpBpl*p质粒编码三种胡萝卜素生成酶，其被工程化以靶向到P. pastonis的过氧化物酶体中，而pGAPZB-EBI* 质粒编码非靶向酶。在将两种质粒转化到巴斯德毕赤酵母中后，选择含有pGAPZB-EpBpl*p质粒的产溶菌酶克隆（称为Omega）并用于进一步优化研究。在所测试的碳源中，葡萄糖导致在复杂和基本培养基中番茄红素产生的最高水平。Omega克隆的分批发酵导致产生4.6 mg-番茄红素/g-DCW，在基本培养基中番茄红素的浓度为73.9 mg/l。首次通过异源表达番茄红素途径酶对非胡萝卜素酵母巴斯德毕赤酵母进行了代谢工程改造，获得了73.9 mg/l的番茄红素浓度。这为在商业生产水平上利用巴斯德毕赤酵母开发类胡萝卜素的生物工艺奠定了基础。(C)2009爱思唯尔有限公司保留所有权利。
As an alternative carotenoid producer, non-carotenogenic Pichia pastoris was chosen for a reddish carotenoid lycopene production because it can grow to high cell density without accumulation of ethanol and utilize various classes of organic materials such as methanol as carbon sources. Two synthetic lycopene-pathway plasmids, pGAPZB-EBI* and pGAPZB-EpBpl*p, were designed and constructed. The pGAPZB-EpBpl*p plasmid encoded three carotenogenic enzymes that were engineered to be targeted into peroxisomes of P. pastonis whereas the pGAPZB-EBI* plasmid encoded non-targeted enzymes. After both plasmids were transformed into P. pastoris, the lycopene-producing clone containing the pGAPZB-EpBpl*p plasmid, referred to as Omega, was selected and used for further optimization study. Of the carbon sources tested, glucose resulted in the highest level of lycopene production in complex and minimal media. Batch fermentation of the Omega clone resulted in the production of 4.6 mg-lycopene/g-DCW, with a concentration of 73.9 mg/l of lycopene in minimal medium. For the first time non-carotenogenic yeast P. pastoris was metabolically engineered by heterologously expressing lycopene-pathway enzymes and the lycopene concentration of 73.9 mg/l was obtained. This serves as a basis for the development of biological process for carotenoids using P. pastoris at a commercial production level. (C) 2009 Elsevier Ltd. All rights reserved.