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This rigorous biochemical evidence points to c HET as a core component of de novo B1 biosynthesis; nonetheless, the vast majority of B1-related research and reviews to date make no mention of c HET (or phosphorylated or adenylated forms) and instead describe only synthesis and use of HET(−P), low additions of c HET (plus the pyrimidine precursor HMP) promoted growth (Fig.
1), confirming our hypothesis and revealing that O.
key microorganisms that influence marine primary productivity (Fig. Large black arrows denote core B1 biosynthesis processes.
Cells acquire exogenous thiazole precursor via high (HA) or low affinity (LA) transport systems (with unknown sequence identity), or diffusion based on results shown here (Figs 1, 2, Supplementary Fig. Shorthand compound names are in italics, while shorthand enzyme names are in bold.
All concentration values along the x-axes are in picomolar. coli ∆thi G cells sustain growth using sub-picomolar concentrations of exogenous c HET or B1. coli ∆thi G mutant exhibited no notable growth upon supplied HET up to 10 p M) of exogenous HET are required to sustain growth of E. Mean maximum yields for triplicate cultures are plotted along with their respective standard deviations.
Asterisks denote significant differences (p can use exogenous c HET. coli (Figs 1, 2), equivalent concentrations of HET also sustained growth of the Arabidopsis mutant (Fig. This result highlights a key difference between plants and (aquatic) microbes, in that the latter are equipped to salvage B1 from very low concentrations of exogenous precursors (Figs 1, 2), likely as a result of adaption within an environment where precursor(s) are a community currency circulated between producers and consumers.
RCC4222, also grew on supplied c HET (and HMP) under B1-limiting conditions (Supplementary Fig.
S1), showing that use of exogenous c HET is a more general phenomenon in marine picoeukaryotic phytoplankton, particularly the Prasinophyceae.
coli can synthesize B1 from HET, but only at relatively high extracellular concentrations of HET, which presumably enters the cell via low-affinity transporters and/or diffusion. coli ∆thi G mutant grows on B1-deplete M63 medium using exogenous c HET.For example: (1) ubiquitous bacterioplankton, affiliated with the SAR11 clade, accounting for more than half of microbes in the oligotrophic surface ocean. to use the newly detected precursor(s) found in seawater and produced by de novo B1-synthesizing plankton - strongly suggesting that the compound(s) is thiazole-related.Intrigued that these marine picoeukaryotic phytoplankton potentially use a novel thiazole precursor, we noted with interest that carboxythiazole, 5-(2-hydroxyethyl)-4-methyl-1,3-thiazole-2-carboxylic acid (c HET), specifically phosphorylated c HET (c HET-P), is produced by bacterial thiazole synthase.(A) RCC745 grows when provided different concentrations of c HET (plus 1 n M HMP) or B1 (1 n M; as a positive control).
The addition of c HET also facilitates use of low concentrations (p M) of HMP by RCC745 (Supplementary Fig. (B) In contrast, a RCC745 ∆thi M mutant does not grow on supplied c HET (plus 1 n M HMP).Our results alter this paradigm as c HET is clearly useful for prokaryotic and eukaryotic microorganisms, and moreover is accessible at extremely low concentrations (Figs 1, 2). Our findings also improve understanding of B1 biosynthesis in general - a vital process for life on Earth, and target for industrial and biomedical applications with human impact, e.g.