Pantothenate auxotrophy in Zymomonas mobilis ZM4 is due to a lack of aspartate decarboxylase activity

Gliessman, Jennifer R; Kremer, Timothy; Sangani, Amee A; Jones-Burrage, Sara E; McKinlay, James B.

doi:10.1093/femsle/fnx136

Cited by 9 publications

(6 citation statements)

References 23 publications

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“…In the case of transformation, for the preparation of electrocompetent C. crescentus cells, a modified protocol developed for Zymomonas mobilis (Gliessman et al ., 2017) was followed. C. crescentus was grown to mid‐exponential phase (OD 600 = 0.45) in 100 ml PYE medium, harvested by centrifugation, washed three times in 10 ml of ice‐cold 10% glycerol and resuspended in 1 ml ice‐cold 10% glycerol.…”

Section: Methodsmentioning

confidence: 99%

Five structural genes required for ceramide synthesis in Caulobacter and for bacterial survival

Olea‐Ozuna

Poggio

EdBergström

et al. 2020

Environmental Microbiology

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Summary Sphingolipids are essential and common membrane components in eukaryotic organisms, participating in many important cellular functions. Only a few bacteria are thought to harbour sphingolipids in their membranes, among them the well‐studied α‐proteobacterium Caulobacter crescentus, a model organism for asymmetric cell division and cellular differentiation. Here, we report that C. crescentus wild type produces several molecular species of dihydroceramides, which are not produced in a mutant lacking the structural gene for serine palmitoyltransferase (spt). Whereas growth of a spt‐deficient mutant and wild type are indistinguishable during the exponential phase of growth, survival of the spt‐deficient mutant is much reduced, in comparison with wild type, during stationary phase of growth, especially at elevated temperatures. The structural gene for spt is located within a genomic cluster, comprising another 16 genes and which, like spt, are important for fitness of C. crescentus. Mutants deficient in genes linked to spt by high cofitness were unable to produce dihydroceramide or to survive in stationary phase of growth at elevated temperatures. At least five structural genes are required for dihydroceramide biosynthesis in C. crescentus and sphingolipid biosynthesis is needed for survival of this bacterium and the integrity of its outer membrane.

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Section: Methodsmentioning

confidence: 99%

Five structural genes required for ceramide synthesis in Caulobacter and for bacterial survival

Olea‐Ozuna

Poggio

EdBergström

et al. 2020

Environmental Microbiology

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“…Transformants were screened by PCR and verified by sequencing. Purified plasmids were then transformed into electrocompetent cells of the appropriate Z. mobilis strain as described elsewhere (46). Cellulase and mechanical dispersion of flocs.…”

Section: Methodsmentioning

confidence: 99%

Cell Aggregation and Aerobic Respiration Are Important for Zymomonas mobilis ZM4 Survival in an Aerobic Minimal Medium

Jones-Burrage

Kremer

McKinlay

2019

Appl Environ Microbiol

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Zymomonas mobilis produces ethanol from glucose near the theoretical maximum yield, making it a potential alternative to the yeast Saccharomyces cerevisiae for industrial ethanol production. A potentially useful industrial feature is the ability to form multicellular aggregates called flocs, which can settle quickly and exhibit higher resistance to harmful chemicals than single cells. While spontaneous floc-forming Z. mobilis mutants have been described, little is known about the natural conditions that induce Z. mobilis floc formation or about the genetic factors involved. Here we found that wild-type Z. mobilis forms flocs in response to aerobic growth conditions but only in a minimal medium. We identified a cellulose synthase gene cluster and a single diguanylate cyclase that are essential for both floc formation and survival in a minimal aerobic medium. We also found that NADH dehydrogenase 2, a key component of the aerobic respiratory chain, is important for survival in a minimal aerobic medium, providing a physiological role for this enzyme, which has previously been found to be disadvantageous in a rich aerobic medium. Supplementation of the minimal medium with vitamins also promoted survival but did not inhibit floc formation. IMPORTANCE The bacterium Zymomonas mobilis is best known for its anaerobic fermentative lifestyle, in which it converts glucose into ethanol at a yield surpassing that of yeast. However, Z. mobilis also has an aerobic lifestyle, which has confounded researchers with its attributes of poor growth, accumulation of toxic acetic acid and acetaldehyde, and respiratory enzymes that are detrimental for aerobic growth. Here we show that a major Z. mobilis respiratory enzyme and the ability to form multicellular aggregates, called flocs, are important for survival, but only during aerobic growth in a medium containing a minimum set of nutrients required for growth. Supplements, such as vitamins or yeast extract, promote aerobic growth and, in some cases, inhibit floc formation. We propose that Z. mobilis likely requires aerobic respiration and floc formation in order to survive in natural environments that lack protective factors found in supplements such as yeast extract.

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“…Such a pantothenate auxotrophy has also been described in the bacterial species Zymomonas mobilis and Methylobacterium spp. (24, 25). In these examples, auxotrophy was due to the lack of the corresponding aspartate decarboxylase PanD or one of the enzymes required to synthesize the precursor β-alanine, respectively.…”

Section: Discussionmentioning

confidence: 99%

Pantothenate auxotrophy in a naturally occurring biocontrol yeast

Mejia

Ortiz‐Merino

Lutz

et al. 2022

Preprint

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The genus Hanseniaspora is characterized by some of the smallest genomes among budding yeasts. These fungi are primarily found on plant surfaces and in fermented products and represent promising biocontrol agents against notorious fungal plant pathogens. In this work, we identify a Hanseniaspora meyeri isolate that shows strong antagonism against the plant pathogen Fusarium oxysporum as a pantothenate auxotroph. Furthermore, strong biocontrol activity in vitro required both pantothenate and biotin in the growth medium. We show that the H. meyeri isolate APC 12.1 can obtain the vitamin from plants and other fungi. The underlying reason for the auxotrophy is the lack of key pantothenate biosynthesis genes, but at least six genes encoding putative pantothenate transporters are present in the genome. By constructing and using a Saccharomyces cerevisiae reporter strain, we identified one Hanseniaspora transporter, out of the six candidate proteins, that conferred pantothenate uptake activity to S. cerevisiae. Pantothenate auxotrophy is rare and has only been described in a few bacteria and in S. cerevisiae strains that were isolated from sake. Such auxotrophic strains may seem an unexpected and unlikely choice as potential biocontrol agents, but they may be particularly competitive in their ecological niche and their specific growth requirements are an inherent biocontainment strategy preventing uncontrolled growth in the environment. Auxotrophic strains such as the H. meyeri isolate APC 12.1 may thus represent a new strategy for developing biocontrol agents that will be easier to register than prototrophic strains, which are normally used for such applications.

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Pantothenate auxotrophy in Zymomonas mobilis ZM4 is due to a lack of aspartate decarboxylase activity

Cited by 9 publications

References 23 publications

Five structural genes required for ceramide synthesis in Caulobacter and for bacterial survival

Five structural genes required for ceramide synthesis in Caulobacter and for bacterial survival

Cell Aggregation and Aerobic Respiration Are Important for Zymomonas mobilis ZM4 Survival in an Aerobic Minimal Medium

Pantothenate auxotrophy in a naturally occurring biocontrol yeast

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