Metabolic Rearrangements Causing Elevated Proline and Polyhydroxybutyrate Accumulation During the Osmotic Adaptation Response of Bacillus megaterium

Godard, Thibault; Zühlke, Daniela; Richter, Georg; Wall, Melanie M.; Rohde, Manfred; Riedel, Katharina; Poblete‐Castro, Ignacio; Krull, Rainer; Biedendieck, Rebekka

doi:10.3389/fbioe.2020.00047

Cited by 16 publications

(25 citation statements)

References 122 publications

(136 reference statements)

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“…Again, L-glutamate was the only compatible solute synthesized by salt-stressed stationary phase B. methanolicus MGA3 cells ( Supplementary Figure 1 ). This is consistent with our in silico analysis of the B. methanolicus MGA3 genome sequence ( Heggeset et al, 2012 ), as we did not find genes for osmostress adaptive synthesis of L-proline ( Brill et al, 2011 ; Schroeter et al, 2013 ; Godard et al, 2020 ), ectoine/hydroxyectoine ( Czech et al, 2018 ), trehalose ( Wolf et al, 2003 ), or for the choline-dependent synthesis of glycine betaine ( Boch et al, 1996 ).…”

Section: Resultssupporting

confidence: 91%

“…This B. subtilis strain has previously been intensively studied with respect to its salt tolerance and the molecular mechanisms underlying this trait ( Boch et al, 1994 ; Hoffmann and Bremer, 2016 , 2017 ). B. subtilis , like other Bacillus species ( Bursy et al, 2007 ; Schroeter et al, 2013 ; Godard et al, 2020 ), synthesizes large quantities of the compatible solute L-proline to counteract high salinity induced osmotic stress ( Whatmore et al, 1990 ; Brill et al, 2011 ). In keeping with the data documented in Figure 1A , we found that B. methanolicus MGA3 is, in comparison with B. subtilis JH642, not particularly resistant to salt stress ( Figure 1B ).…”

Section: Resultsmentioning

confidence: 99%

“…Members of the genus Bacillus make effective use of compatible solutes as protectants against high osmolarity induced cellular challenges ( Hoffmann and Bremer, 2016 ). Depending on the species, Bacilli synthesize the compatible solutes L-glutamate, L-proline, or ectoine/hydroxyectoine (or a combination thereof) ( Whatmore et al, 1990 ; Kuhlmann and Bremer, 2002 ; Bursy et al, 2007 ; Brill et al, 2011 ; Schroeter et al, 2013 ; Godard et al, 2020 ), and they can produce the osmostress protectant glycine betaine from prior imported choline ( Boch et al, 1994 , 1996 ). In addition, and as studied in detail in B. subtilis , several high-affinity uptake systems for compatible solutes operate in high-osmolarity stressed cells ( Hoffmann and Bremer, 2017 ; Teichmann et al, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

“…In our study, we discovered that in comparison with Bacilli already used as chassis for industrial-scale production processes (e.g., B. subtilis , B. licheniformis , and B. megaterium ), B. methanolicus possesses a rather restricted ability to withstand sustained high osmolarity incurred stress. This property is probably linked to its exclusive synthesis of the moderately effective compatible solute L-glutamate, while B. subtilis , B. licheniformis , and B. megaterium all synthesize L-proline as their dominant compatible solute ( Whatmore et al, 1990 ; Brill et al, 2011 ; Schroeter et al, 2013 ; Godard et al, 2020 ). Surprisingly, we observed that the B. methanolicus strain MGA3 cannot achieve osmostress resistance through import of compatible solutes or through the uptake and hydrolysis of proline-containing peptides, processes that make major contributions to the development of osmostress tolerance by B. subtilis ( Hoffmann and Bremer, 2016 , 2017 ).…”

Section: Introductionmentioning

confidence: 99%

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Enhanced Glutamate Synthesis and Export by the Thermotolerant Emerging Industrial Workhorse Bacillus methanolicus in Response to High Osmolarity

et al. 2021

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The thermotolerant methylotroph Bacillus methanolicus MGA3 was originally isolated from freshwater marsh soil. Due to its ability to use methanol as sole carbon and energy source, B. methanolicus is increasingly explored as a cell factory for the production of amino acids, fine chemicals, and proteins of biotechnological interest. During high cell density fermentation in industrial settings with the membrane-permeable methanol as the feed, the excretion of low molecular weight products synthesized from it will increase the osmotic pressure of the medium. This in turn will impair cell growth and productivity of the overall biotechnological production process. With this in mind, we have analyzed the core of the physiological adjustment process of B. methanolicus MGA3 to sustained high osmolarity surroundings. Through growth assays, we found that B. methanolicus MGA3 possesses only a restricted ability to cope with sustained osmotic stress. This finding is consistent with the ecophysiological conditions in the habitat from which it was originally isolated. None of the externally provided compatible solutes and proline-containing peptides affording osmostress protection for Bacillus subtilis were able to stimulate growth of B. methanolicus MGA3 at high salinity. B. methanolicus MGA3 synthesized the moderately effective compatible solute L-glutamate in a pattern such that the cellular pool increased concomitantly with increases in the external osmolarity. Counterintuitively, a large portion of the newly synthesized L-glutamate was excreted. The expression of the genes (gltAB and gltA2) for two L-glutamate synthases were upregulated in response to high salinity along with that of the gltC regulatory gene. Such a regulatory pattern of the system(s) for L-glutamate synthesis in Bacilli is new. Our findings might thus be generally relevant to understand the production of the osmostress protectant L-glutamate by those Bacilli that exclusively rely on this compatible solute for their physiological adjustment to high osmolarity surroundings.

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Section: Resultssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Enhanced Glutamate Synthesis and Export by the Thermotolerant Emerging Industrial Workhorse Bacillus methanolicus in Response to High Osmolarity

et al. 2021

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“…In contrast much effort has been placed on optimizing cultivation conditions of environmental isolates of P. megaterium to increase PHB production with mainly molasses as carbon source, resulting in almost 70% PHB of cell dry weight (Gouda et al 2001 ; Rodríguez-Contreras et al 2013 ). For the well-known and genome sequenced strain DSM319, Godard et al ( 2020 ) observed that the PHB content increased 5-fold to almost 30 % of cell dry weight under high salt conditions (Godard et al 2020 ). Further, the production of functionalized PHB granules provides an exciting new application for P. megaterium .…”

Section: Production Of Biopolymers Using P Megaterium : Polyhydroxybutyrate (Phb)mentioning

confidence: 99%

The “beauty in the beast”—the multiple uses of Priestia megaterium in biotechnology

Biedendieck

Knuuti

Jahn

2021

Appl Microbiol Biotechnol

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Over 30 years, the Gram-positive bacterium Priestia megaterium (previously known as Bacillus megaterium) was systematically developed for biotechnological applications ranging from the production of small molecules like vitamin B12, over polymers like polyhydroxybutyrate (PHB) up to the in vivo and in vitro synthesis of multiple proteins and finally whole-cell applications. Here we describe the use of the natural vitamin B12 (cobalamin) producer P. megaterium for the elucidation of the biosynthetic pathway and the subsequent systematic knowledge-based development for production purposes. The formation of PHB, a natural product of P. megaterium and potential petro-plastic substitute, is covered and discussed. Further important biotechnological characteristics of P. megaterium for recombinant protein production including high protein secretion capacity and simple cultivation on value-added carbon sources are outlined. This includes the advanced system with almost 30 commercially available expression vectors for the intracellular and extracellular production of recombinant proteins at the g/L scale. We also revealed a novel P. megaterium transcription-translation system as a complementary and versatile biotechnological tool kit. As an impressive biotechnology application, the formation of various cytochrome P450 is also critically highlighted. Finally, whole cellular applications in plant protection are completing the overall picture of P. megaterium as a versatile giant cell factory. Key points • The use of Priestia megaterium for the biosynthesis of small molecules and recombinant proteins through to whole-cell applications is reviewed. • P. megaterium can act as a promising alternative host in biotechnological production processes.

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Bacterial Community Structure and Function in Acid Soil Ecosystem

Barooah

Hazarika

Deka

2022

Advances in Agricultural and Industrial Microbiology

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Metabolic Rearrangements Causing Elevated Proline and Polyhydroxybutyrate Accumulation During the Osmotic Adaptation Response of Bacillus megaterium

Cited by 16 publications

References 122 publications

Enhanced Glutamate Synthesis and Export by the Thermotolerant Emerging Industrial Workhorse Bacillus methanolicus in Response to High Osmolarity

Enhanced Glutamate Synthesis and Export by the Thermotolerant Emerging Industrial Workhorse Bacillus methanolicus in Response to High Osmolarity

The “beauty in the beast”—the multiple uses of Priestia megaterium in biotechnology

Bacterial Community Structure and Function in Acid Soil Ecosystem

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