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A Yeast-Based Biosensor for Screening of Short- and Medium-Chain Fatty Acid Production
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-10-19 00:00:00 , DOI: 10.1021/acssynbio.8b00309 Leonie Baumann 1 , Arun S. Rajkumar 2 , John P. Morrissey 2 , Eckhard Boles 1 , Mislav Oreb 1
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-10-19 00:00:00 , DOI: 10.1021/acssynbio.8b00309 Leonie Baumann 1 , Arun S. Rajkumar 2 , John P. Morrissey 2 , Eckhard Boles 1 , Mislav Oreb 1
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Short- and medium-chain fatty acids (SMCFA) are important platform chemicals currently produced from nonsustainable resources. The engineering of microbial cells to produce SMCFA, however, lacks high-throughput methods to screen for best performing cells. Here, we present the development of a whole-cell biosensor for easy and rapid detection of SMCFA. The biosensor is based on a multicopy yeast plasmid containing the SMCFA-responsive PDR12 promoter coupled to GFP as the reporter gene. The sensor detected hexanoic, heptanoic and octanoic acid over a linear range up to 2, 1.5, and 0.75 mM, respectively, but did not show a linear response to decanoic and dodecanoic acid. We validated the functionality of the biosensor with culture supernatants of a previously engineered Saccharomyces cerevisiae octanoic acid producer strain and derivatives thereof. The biosensor signal correlated strongly with the octanoic acid concentrations as determined by gas chromatography. Thus, this biosensor enables the high-throughput screening of SMCFA producers and has the potential to drastically speed up the engineering of diverse SMCFA producing cell factories.
中文翻译：
用于筛选短链和中链脂肪酸生产的基于酵母的生物传感器
短链和中链脂肪酸（SMCFA）是目前由不可持续资源生产的重要平台化学品。然而，用于生产SMCFA的微生物细胞工程缺乏用于筛选性能最佳的细胞的高通量方法。在这里，我们介绍了一种用于简单快速检测SMCFA的全细胞生物传感器的开发。该生物传感器基于多拷贝酵母质粒，该质粒包含与GFP偶联的SMCFA反应性PDR12启动子作为报告基因。该传感器分别在高达2、1.5和0.75 mM的线性范围内检测到己酸，庚酸和辛酸，但对癸酸和十二烷酸没有线性响应。我们用先前设计的酿酒酵母培养上清液验证了生物传感器的功能辛酸生产菌株及其衍生物。生物传感器信号与通过气相色谱法测定的辛酸浓度高度相关。因此，这种生物传感器可以对SMCFA生产者进行高通量筛选，并具有极大地加快各种SMCFA生产细胞工厂的工程设计的潜力。
更新日期：2018-10-19
中文翻译：
用于筛选短链和中链脂肪酸生产的基于酵母的生物传感器
短链和中链脂肪酸（SMCFA）是目前由不可持续资源生产的重要平台化学品。然而，用于生产SMCFA的微生物细胞工程缺乏用于筛选性能最佳的细胞的高通量方法。在这里，我们介绍了一种用于简单快速检测SMCFA的全细胞生物传感器的开发。该生物传感器基于多拷贝酵母质粒，该质粒包含与GFP偶联的SMCFA反应性PDR12启动子作为报告基因。该传感器分别在高达2、1.5和0.75 mM的线性范围内检测到己酸，庚酸和辛酸，但对癸酸和十二烷酸没有线性响应。我们用先前设计的酿酒酵母培养上清液验证了生物传感器的功能辛酸生产菌株及其衍生物。生物传感器信号与通过气相色谱法测定的辛酸浓度高度相关。因此，这种生物传感器可以对SMCFA生产者进行高通量筛选，并具有极大地加快各种SMCFA生产细胞工厂的工程设计的潜力。
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