One major facilitator superfamily transporter is responsible for propionic acid tolerance in Pseudomonas putida KT2440.
One major facilitator superfamily transporter is responsible for propionic acid tolerance in Pseudomonas putida KT2440.
复制标题
一种主要的促进超家族转运蛋白负责恶臭假单胞菌 KT2440 的丙酸耐受性。
DOI：
10.1111/1751-7915.13597
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发表时间：
2021-03
影响因子：
5.7
通讯作者：
Ma Y
中科院分区：
文献类型：
--
作者：
Ma C;Mu Q;Xue Y;Xue Y;Yu B;Ma Y
丙酸（PA）广泛用作食品防腐剂和农业和制药工业的化学中间体。利用生物质生产环保型PA已被认为是传统石油化工路线的替代方案。然而，由于PA是细胞生长的强抑制剂，生物技术宿主不仅应该能够产生化合物，而且宿主应该是健壮的。在这项研究中，我们确定了关键PA耐受因子恶臭假单胞菌KT 2440菌株在PA胁迫的存在或不存在的比较转录分析。通过实验验证了所鉴定的主要易化超家族（MFS）转运蛋白基因簇PP_1271、PP_1272和PP_1273参与恶臭假单胞菌菌株的PA耐受性。该簇的过表达改善了PA生产菌株对PA的耐受性，这对于进一步工程化该稳健平台不仅用于PA合成而且用于其他弱酸的生产是有用的。在这项研究中，一个主要的易化超家族转运蛋白（MFS 2，PP_1271-PP_1273的位点标签），而不是MFS 1（PP_0503的位点标签），有利于恶臭假单胞菌菌株KT 2440的丙酸盐耐受性。因此，本研究揭示了恶臭假单胞菌菌株中负责PA耐受性的关键基因，并为进一步工程改造用于PA生产的稳健菌株提供了有用的信息。
Propionic acid (PA) has been widely used as a food preservative and chemical intermediate in the agricultural and pharmaceutical industries. Environmental and friendly biotechnological production of PA from biomass has been considered as an alternative to the traditional petrochemical route. However, because PA is a strong inhibitor of cell growth, the biotechnological host should be not only able to produce the compound but the host should be robust. In this study, we identified key PA tolerance factors in Pseudomonas putida KT2440 strain by comparative transcriptional analysis in the presence or absence of PA stress. The identified major facilitator superfamily (MFS) transporter gene cluster of PP_1271, PP_1272 and PP_1273 was experimentally verified to be involved in PA tolerance in P. putida strains. Overexpression of this cluster improved tolerance to PA in a PA producing strain, what is useful to further engineer this robust platform not only for PA synthesis but for the production of other weak acids. In this study, one major facilitator superfamily transporter (MFS2, locus tags of PP_1271‐PP_1273), but not MFS1 (locus tag of PP_0503), is beneficial to propionate tolerance in Pseudomonas putida strain KT2440. Thus, this study revealed the key genes responsible for PA tolerance in P. putida strains and provided the useful information to further engineer robust strains for PA production.