Functionalized poly(3-hydroxybutyric acid) bodies as new in vitro biocatalysts

Stenger, Benjamin; Gerber, Adrian; Bernhardt, Rita; Hannemann, Frank

doi:10.1016/j.bbapap.2017.08.010

Cited by 4 publications

(3 citation statements)

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“…One recent study showed that the mammalian cytochrome P450 CYP11A1 could be immobilized and purified with PHB granules produced in P. megaterium , thereby circumventing the problem of low stability of recombinantly produced cytochromes. Here, CYP11A1 was readily localized in the phospholipid monolayer of the PHB granule in its native form verified by denaturing PAGE (Stenger et al 2018 ). Another study showed that the IgG binding domain of Protein A from Staphylococcus aureus (ZZ domain) could be produced, purified, and presented on PHB granules when fused to PhaC in P. megaterium .…”

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

confidence: 79%

“…In addition, E. coli -based whole-cell systems using CYP107DY1 from P. megaterium strain QM B 1551 (Milhim et al 2016 ) and CYP102A1 from strain DSM32 (Chu et al 2016 ) have been reported. Finally, membrane-bound mammalian P450 CYP11A1 was recombinantly produced in P. megaterium (Stenger et al 2018 ).…”

Section: Production Of Biotechnological Important Proteins: Multiple Cytochrome P450 Suitable For Whole-cell Transformationsmentioning

confidence: 99%

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

confidence: 79%

Section: Production Of Biotechnological Important Proteins: Multiple Cytochrome P450 Suitable For Whole-cell Transformationsmentioning

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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“…Acetoacetyl-CoA is subsequently reduced to d -3-hydroxybutyryl-CoA by the acetoacetyl-CoA reductase PhaB, and finally, the polymerization reaction is catalyzed by the PHB synthase PhaC. − PHAs have been widely evaluated as an environmentally friendly surrogate for petroleum-based plastics as the bioplastic can be sustainably synthesized in natural producers or engineered bacteria and exhibits beneficial properties including good biocompatibility, high biodegradability, and nontoxicity . Under appropriate growth conditions, synthesized PHA makes up up to 90% of the cell dry weight and accumulates in the cytoplasm as spherical particles with a size of 100–500 nm. , Besides the hydrophobic PHA core, the granules are surrounded by a protein layer. , This layer is composed of different PHA-associated proteins including PhaC, different phasins (e.g., PhaP or PhaF), a depolymerase, and other regulatory and structural proteins. − Based on this observation, PHA granules were further used to develop a versatile in vivo protein immobilization and display technology. ,,− In most cases, accordingly engineered E. coli strains, harboring the essential PHB biosynthesis genes, are applied for POI in vivo immobilization. For this purpose, the synthase PhaC can be employed as a versatile anchor protein, because it tolerates POI fusions at its N- and C-termini.…”

Section: Polyhydroxyalkanoate-based Systems and Viruslike Particlesmentioning

confidence: 99%

Emerging Solutions for in Vivo Biocatalyst Immobilization: Tailor-Made Catalysts for Industrial Biocatalysis

Ölçücü

Klaus

Jaeger

et al. 2021

ACS Sustainable Chem. Eng.

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In industry, enzymes are often immobilized to generate more stable enzyme preparations that are easier to store, handle, and recycle for repetitive use. Traditionally, enzymes are bound to inorganic carrier materials, which requires caseto-case optimization and incurs additional labor and costs. Therefore, with the advent of rational protein design strategies as part of bottom-up synthetic biology approaches, numerous immobilization methods have been developed that enable the one-step production and immobilization of enzymes onto biogenic carrier materials often directly within the production host, which we here refer to as in vivo immobilization. As a result, nano-to micro-meter-sized functionalized biomaterials, or biologically produced enzyme immobilizates, are obtained that can directly be used for synthetic purposes. In this Perspective, we provide an overview over established and recently emerging in vivo enzyme immobilization methods, with special emphasis on their applicability for (industrial) biocatalysis. For each approach, we present fundamental working principles as well as advantages and limitations guiding future research avenues toward sustainable applications in the bioindustry.

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