Controlled production of polysaccharides–exploiting nutrient supply for levan and heteropolysaccharide formation in Paenibacillus sp.

Rütering, Marius; Schmid, Jochen; Rühmann, Broder; Schilling, M.; Sieber, Volker

doi:10.1016/j.carbpol.2016.04.074

Cited by 57 publications

(36 citation statements)

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“…Xanthomonas campestris pv. campestris LMG 8031 (Xcc) (BCCM, Gent, Belgium) and deletion variants thereof were cultured at 30 °C in MM1 P100 as previously described (Rütering, Schmid, Rühmann, Schilling, & Sieber, 2016) containing 30 g L -1 glucose as carbon source. The fermentative production of xanthan and engineered variants thereof were carried out in a 2 L-benchtop fermenter (Biostat B plus, Sartorius AG, Germany) equipped with two 6 blade-stirrers over 48 h with a controlled pH of 6.8 and a pO2 of 30% saturation.…”

Section: Fermentative Cultivation Of Strains and Production Of Xanthanmentioning

confidence: 99%

In-depth rheological characterization of genetically modified xanthan-variants

Gansbiller

Schmid

Sieber

2019

Carbohydrate Polymers

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Section: Fermentative Cultivation Of Strains and Production Of Xanthanmentioning

confidence: 99%

In-depth rheological characterization of genetically modified xanthan-variants

Gansbiller

Schmid

Sieber

2019

Carbohydrate Polymers

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“…Their extensive enzymatic capabilities to degrade complex carbohydrates and to produce and tolerate high levels of commercially relevant chemicals like 2,3-butanediol, for example, make Paenibacilli an interesting genus for the fermentative production of this and other platform chemicals from renewable resources ( 30 ). Critically, they produce a class of still underappreciated but highly promising EPSs possessing antioxidant activity and outstanding rheological properties, qualifying them for applications in therapeutics or as thickeners ( 31–35 ). Grady et al ( 36 ) recently reviewed the potential of Paenibacilli in agriculture and industrial biotechnology in detail.…”

Section: Introductionmentioning

confidence: 99%

Tailor-made exopolysaccharides—CRISPR-Cas9 mediated genome editing in Paenibacillus polymyxa

et al. 2017

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Application of state-of-the-art genome editing tools like CRISPR-Cas9 drastically increase the number of undomesticated micro-organisms amenable to highly efficient and rapid genetic engineering. Adaptation of these tools to new bacterial families can open up entirely new possibilities for these organisms to accelerate as biotechnologically relevant microbial factories, also making new products economically competitive. Here, we report the implementation of a CRISPR-Cas9 based vector system in Paenibacillus polymyxa, enabling fast and reliable genome editing in this host. Homology directed repair allows for highly efficient deletions of single genes and large regions as well as insertions. We used the system to investigate the yet undescribed biosynthesis machinery for exopolysaccharide (EPS) production in P. polymyxa DSM 365, enabling assignment of putative roles to several genes involved in EPS biosynthesis. Using this simple gene deletion strategy, we generated EPS variants that differ from the wild-type polymer not only in terms of monomer composition, but also in terms of their rheological behavior. The developed CRISPR-Cas9 mediated engineering approach will significantly contribute to the understanding and utilization of socially and economically relevant Paenibacillus species and extend the polymer portfolio.

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“…As shown in Table 1 , fully sequenced P. polymyxa strains have a single copy of the levansucrase gene, which has 95-97% protein sequence similarity to that of P. polymyxa DSM 365. Levansucrase, which is responsible for levan EPS biosynthesis in P. polymyxa (36, 37), was targeted for inactivation. The results from the present study are discussed below.…”

Section: Discussionmentioning

confidence: 99%

Molecular inactivation of exopolysaccharide biosynthesis inPaenibacillus polymyxaDSM 365 for enhanced 2,3-butanediol production

Okonkwo

Ujor

Ezeji

2018

Preprint

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25Formation of Exopolysaccharides (EPS) during 2,3-butanediol (2,3-BD) fermentation by 26 Paenibacillus polymyxa decreases 2,3-BD yield, increases medium viscosity and impacts 2,3-BD 27 downstream processing. Therefore, additional purification steps are required to rid the fermentation 28 broth of EPS prior to 2,3-BD purification, which adds to the production cost. To eliminate EPS 29 production during 2,3-BD fermentation, we explored a metabolic engineering strategy to disable the 30 EPS production pathway of P. polymyxa, thereby increasing 2,3-BD yield and productivity. The 31 levansucrase gene which encodes levansucrase, the enzyme responsible for EPS biosynthesis in P. 32 polymyxa, was successfully disrupted. The resulting P. polymyxa levansucrase null mutant showed 33 34% and 54% increases in growth with 6.4-and 2.4-folds decrease in EPS formation in sucrose and 34 glucose cultures, respectively. The observed decrease in EPS formation by the levansucrase null 35 mutant may account for the 27% and 4% increase in 2,3-BD yield, and 4% and 128% increases in 36 2,3-BD productivity when grown on sucrose and glucose media, respectively. Genetic stability of 37 the levansucrase null mutant was further evaluated. Interestingly, the levansucrase null mutant 38 remained genetically stable over fifty generations with no observable decrease in growth and 2,3-39 BD formation with or without antibiotic supplementations. Collectively, our results show that P. 40 polymyxa levansucrase null mutant has potential for improving 2,3-BD yield, and ultimately, the 41 economics of large-scale microbial 2,3-BD production. 42 43 44 polysaccharide polymerase 45 46 47 48 49Considering the finite nature of fossil fuels, recurrent instability in oil price and the 50 environmental concerns associated with oil consumption, there is an urgent need to develop 51 sustainable alternatives to fossil fuels and their derivatives. Over the past few decades, significant 52 attention has been devoted to the development of alternative sources of fuels and chemicals. 2,3-53 Butanediol (2,3-BD) is an industrial platform chemical that is generated via cracking of petroleum-54 derived hydrocarbons (e.g. butenes). 2,3-BD has wide industrial applications. For instance, 2,3-BD 55 can be used as a feedstock chemical in the production of 1,3-butadiene (1,3-BD), the monomer from 56 which synthetic rubber is produced (1, 2). Also, 2,3-BD can be used as a feedstock for producing 57 methyl ethyl ketone (MEK), a fuel additive which has a higher heat of combustion than ethanol, and 58 as a solvent from which resins and lacquers can be produced (1, 2). Additionally, 2,3-BD has 59 massive potential as a feedstock for the synthesis of a host of numerous pharmaceuticals, cosmetics, 60 paints, and food preservatives (3, 4). 61 Several microorganisms have been shown to possess the metabolic machinery to convert 62 carbohydrates to 2,3-BD. However, 2,3-BD is produced via a mixed acid fermentation pathway 63 where other products such as ethanol, acetoin, lact...

show abstract

Controlled production of polysaccharides–exploiting nutrient supply for levan and heteropolysaccharide formation in Paenibacillus sp.

Cited by 57 publications

References 50 publications

In-depth rheological characterization of genetically modified xanthan-variants

In-depth rheological characterization of genetically modified xanthan-variants

Tailor-made exopolysaccharides—CRISPR-Cas9 mediated genome editing in Paenibacillus polymyxa

Molecular inactivation of exopolysaccharide biosynthesis inPaenibacillus polymyxaDSM 365 for enhanced 2,3-butanediol production

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