Department of Biosciences and Bioengineering, Indian Institute of Technology-Bombay, Powai, Mumbai 400 076, India.
Present address: Presently affiliated to TCR Therapeutics, Inc., 100 Binney Street, Cambridge, MA 02142, USA.
Microbiology (Reading). 2022 Aug;168(8). doi: 10.1099/mic.0.001206.
Hierarchical utilization of substrate by microbes (utilization of simple carbon sources prior to complex ones) poses a major limitation to the efficient remediation of aromatic pollutants. Aromatic compounds, being complex and reduced in nature, appear to be a deferred choice as the carbon source in the presence of a plethora of simple organic compounds in the environment. The soil bacterium CSV86 displays a unique carbon source utilization hierarchy. It preferentially utilizes aromatics over glucose and co-metabolizes them with succinate or pyruvate (Basu ., 2006, , 72 : 22226-2230). In the present study, the substrate utilization hierarchy for strain CSV86 was tested for additional simple carbon sources such as glycerol, acetate, and tri-carboxylic acid (TCA) cycle intermediates like α-ketoglutarate and fumarate. When grown on a mixture of aromatics (benzoate or naphthalene) plus glycerol, the strain displayed a diauxic growth profile with significantly high activity of aromatic utilization enzymes (catechol 1,2- or 2,3-dioxygenase, respectively) in the first-log phase. This suggests utilization of aromatics in the first-log phase followed by glycerol in the second-log phase. On a mixture of an aromatic plus organic acid (acetate, α-ketoglutarate or fumarate), the strain displayed a monoauxic growth profile, indicating co-metabolism. Interestingly, the presence of glycerol, acetate, α-ketoglutarate or fumarate does not repress metabolism/utilization of the aromatic. Thus, the substrate utilization hierarchy of strain CSV86 is aromatics=organic acids>glucose/glycerol, which is unique as compared to other species, where degradation of aromatics is repressed by glycerol, glucose, acetate or organic acids, including TCA cycle intermediates. This novel substrate utilization hierarchy appears to be a global metabolic phenomenon in strain CSV86, thus implying it to be an ideal host for metabolic engineering as well as for its potential application in bioremediation.
微生物对基质的层级利用(先利用简单的碳源,再利用复杂的碳源)会极大地限制芳香族污染物的有效修复。芳香族化合物性质复杂且还原程度高,在环境中存在大量简单有机化合物的情况下,它们似乎不是首选的碳源。土壤细菌 CSV86 表现出独特的碳源利用层次。它优先利用芳香族化合物而不是葡萄糖,并与琥珀酸盐或丙酮酸共代谢它们(Basu.,2006,72:22226-2230)。在本研究中,测试了 CSV86 菌株对其他简单碳源(如甘油、乙酸盐和三羧酸 (TCA) 循环中间体如α-酮戊二酸和富马酸)的底物利用层次。当在芳香族化合物(苯甲酸或萘)和甘油的混合物上生长时,该菌株表现出双相生长模式,在第一对数生长期具有明显高的芳香族利用酶(分别为儿茶酚 1,2-或 2,3-加氧酶)活性。这表明在第一对数生长期利用芳香族化合物,然后在第二对数生长期利用甘油。在芳香族化合物和有机酸(乙酸盐、α-酮戊二酸或富马酸)的混合物上,该菌株表现出单相生长模式,表明共代谢。有趣的是,甘油、乙酸盐、α-酮戊二酸或富马酸的存在不会抑制芳香族化合物的代谢/利用。因此,CSV86 菌株的底物利用层次是芳香族化合物=有机酸>葡萄糖/甘油,与其他物种相比是独特的,在其他物种中,甘油、葡萄糖、乙酸盐或有机酸(包括 TCA 循环中间体)会抑制芳香族化合物的降解。这种新型的底物利用层次似乎是 CSV86 菌株的一种全局代谢现象,因此暗示它是代谢工程的理想宿主,以及在生物修复中的潜在应用。