Improving the penicillin fermentation by modelling and optimising its metabolic network and transporterome

通过建模和优化青霉素的代谢网络和转运体来改善青霉素发酵

基本信息

批准号：
BB/R014744/1
负责人：
Douglas Kell
金额：
$ 54.65万
依托单位：
University of Liverpool
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2018
资助国家：
英国
起止时间：
2018 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FR014744%2F1
关键词：
Improving penicillin fermentation modelling optimising

项目摘要

Penicillin was famously (re)discovered by Alexander Fleming in 1928, and developed into an initial fermentation process by Florey, Chain and Heatley during the early 1940s. Despite increasing antimicrobial resistance, it remains one of the UK fermentation industry's top products (often combined with the penicillinase inhibitor clavulanic acid in products such as Augmentin). However, in terms of the efficiency of conversion of carbon in the sugar feedstock to carbon in the penicillin product it is a lousy fermentation as this efficiency is no more than 10%. Many other fermentations, such as that producing monosodium glutamate, have a carbon conversion efficiency approaching 100%. Consequently, there is much room for improvement, and for making the penicillin process more competitive economically. As with the design of engineering artefacts such as the Boeing 777, what is needed is a mathematical model of the metabolic network of the penicillin producer, P. chrysogenum. The amount that each gene is expressed tells us what is going on, and is known as the transcriptome. To this end, transcriptome data FROM THE PRODUCTION STRAIN ITSELF will be made available by GSK. From the (known) genome sequence we can produce a computer version of the metabolic network for analysis. Importantly, the media used for the penicillin process are fully defined, which makes it feasible to do this modelling. Having produced the model, we can predict, initially qualitatively, what molecules it will produce, and these will be measured experimentally on extracts of cells and medium provided by GSK. The combination of the model and the transcriptome allows us to calculate all the fluxes, both to product and to non-profitable places. This will help determine which changes in the genetic make-up of the Penicillium fungi are most likely to lead to a higher carbon conversion efficiency. These changes will be made (by GSK) and tested on the new production strains. The ability to do these analyses on cell extracts and inside a computer means that while we have access to the data we neither have nor need access to the proprietary production strains themselves.Importantly, we shall curate all of the data in a suitable database.

青霉素是由亚历山大·弗莱明（Alexander Fleming）在1928年发现的，并于1940年代初被弗洛里（Florey），链条和希特利（Heatley）发酵成最初的发酵过程。尽管抗菌素耐药性增加，但它仍然是英国发酵行业的顶级产品之一（在增强素等产品中通常与青霉素酶抑制剂clavulanic结合使用）。然而，就糖原料中碳转化为碳碳中的碳中的碳的效率而言，这是一种糟糕的发酵，因为这种效率不超过10％。许多其他发酵，例如产生谷氨酸单钠的发酵，其碳转化效率接近100％。因此，有很大的改进空间可以使青霉素过程在经济上更具竞争力。与波音777这样的工程伪像的设计一样，需要的是青霉素生产商的代谢网络的数学模型。每个基因表示的数量告诉我们发生了什么，被称为转录组。为此，GSK将提供来自生产应变本身的转录组数据。从（已知的）基因组序列中，我们可以生成代谢网络的计算机版本进行分析。重要的是，用于青霉素过程的介质已完全定义，这使得进行此建模使其可行。产生了该模型后，我们可以在定性上预测其将产生的分子，这些分子将在实验上以GSK提供的细胞提取物和培养基提取。该模型和转录组的组合使我们能够计算出所有通量，包括产品和非营利位置。这将有助于确定真菌遗传组成的哪些变化最有可能导致较高的碳转化效率。这些更改将（由GSK）进行，并对新的生产菌株进行了测试。对细胞提取物进行这些分析和计算机内部进行这些分析的能力意味着，尽管我们可以访问数据，但我们既不需要访问专有生产菌株。