利用磷酸葡萄糖异构酶缺陷的酿酒酵母生产 d-葡萄糖酸

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Mervi Toivari, Maija-Leena Vehkomäki, Laura Ruohonen, Merja Penttilä, Marilyn G. Wiebe
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引用次数: 0

摘要

d-Glucaric acid 是一种潜在的生物基平台化学品。以前主要是大肠杆菌,但也有酵母菌(Saccharomyces cerevisiae)和酿酒酵母(Pichia pastoris)通过肌醇将 d-葡萄糖转化为 d-葡萄糖酸。酵母菌株产量低的一个原因是糖酵解通量大。因此,为了减少 d-葡萄糖转化为生物质的通量,并提高 d-葡萄糖酸的产量,我们在磷酸葡萄糖异构酶缺乏(Pgi1p-deficient)的酿酒酵母菌株中引入了四步 d-葡萄糖酸途径。高浓度的 d-葡萄糖对 Pgi1p 缺乏菌株有毒性,因此研究了各种喂养策略和聚合底物的使用。统一标记的 13C 葡萄糖证实了 d-葡萄糖向 d-葡萄糖酸的转化。在使用脉冲式 d-果糖和乙醇的批次生物反应器培养中,产生了 1.3 克 d-葡糖酸 L-1。在限氮条件下,d-葡糖酸滴度(0.71 g d-葡糖酸 L-1)较低,但产量(0.23 g d-葡糖酸[g d-葡萄糖消耗量]-1)却是迄今为止从酵母中获得的最高产量之一。肌醇的积累表明肌醇加氧酶的活性受到了限制,因此有可能获得更高的产量。谷氨酸酵母中的 Pgi1p 缺陷提供了一种方法,与其他已报道的改造和生物加工策略相结合,将促进高产 d-葡糖酸酵母菌株的发展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Production of d-glucaric acid with phosphoglucose isomerase-deficient Saccharomyces cerevisiae

Production of d-glucaric acid with phosphoglucose isomerase-deficient Saccharomyces cerevisiae

d-Glucaric acid is a potential biobased platform chemical. Previously mainly Escherichia coli, but also the yeast Saccharomyces cerevisiae, and Pichia pastoris, have been engineered for conversion of d-glucose to d-glucaric acid via myo-inositol. One reason for low yields from the yeast strains is the strong flux towards glycolysis. Thus, to decrease the flux of d-glucose to biomass, and to increase d-glucaric acid yield, the four step d-glucaric acid pathway was introduced into a phosphoglucose isomerase deficient (Pgi1p-deficient) Saccharomyces cerevisiae strain. High d-glucose concentrations are toxic to the Pgi1p-deficient strains, so various feeding strategies and use of polymeric substrates were studied. Uniformly labelled 13C-glucose confirmed conversion of d-glucose to d-glucaric acid. In batch bioreactor cultures with pulsed d-fructose and ethanol provision 1.3 g d-glucaric acid L−1 was produced. The d-glucaric acid titer (0.71 g d-glucaric acid L−1) was lower in nitrogen limited conditions, but the yield, 0.23 g d-glucaric acid [g d-glucose consumed]−1, was among the highest that has so far been reported from yeast. Accumulation of myo-inositol indicated that myo-inositol oxygenase activity was limiting, and that there would be potential to even higher yield. The Pgi1p-deficiency in S. cerevisiae provides an approach that in combination with other reported modifications and bioprocess strategies would promote the development of high yield d-glucaric acid yeast strains.

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来源期刊
Biotechnology Letters
Biotechnology Letters 工程技术-生物工程与应用微生物
CiteScore
5.90
自引率
3.70%
发文量
108
审稿时长
1.2 months
期刊介绍: Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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