Mini review: Recombinant production of tailored bio-pharmaceuticals in different Bacillus strains and future perspectives

Lakowitz, Antonia; Godard, Thibault; Biedendieck, Rebekka; Krull, Rainer

doi:10.1016/j.ejpb.2017.06.008

Cited by 26 publications

(14 citation statements)

References 164 publications

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“…There is little evidence that deletions in ispA increase recombinant protein production per se , except for a report that a multiply deleted strain DB431, with deficiencies in the aprE, npr, epr, mpr, ispA and bpr genes leading to an approximate doubling of the concentration of the single chain antibody scFv compared with the wild-type strain (Lakowitz et al . 2018 ). However, it is not possible to ascribe this increase specifically to a lack of IspA.…”

Section: Intracellular Proteasesmentioning

confidence: 99%

The ins and outs of Bacillus proteases: activities, functions and commercial significance

Harwood

Kikuchi

2021

FEMS Microbiology Reviews

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Because the majority of bacterial species divide by binary fission, and do not have distinguishable somatic and germ-lines cells, they could be considered to be immortal. However, bacteria ‘age’ due to damage to vital cell components such as DNA and proteins. DNA damage can often be repaired using efficient DNA repair mechanisms. However, many proteins have a functional ‘shelf life’; some short lived, others are relatively stable. Specific degradation processes are built into the life span of proteins whose activities are required to fulfil a specific function during a prescribed period of time (e.g. cell cycle, differentiation process, stress response). In addition, proteins that are irreparably damaged or that have come to the end of their functional lifespan need to be removed by quality control proteases. Other proteases are involved in performing a variety of specific functions that can be broadly divided in to three categories: processing, regulation and feeding. This review presents a systematic account of the proteases of Bacillus subtilis and their activities. It reviews the proteases found in, or associated with, the cytoplasm, the cell membrane, the cell wall and the external milieu. Where known, the impacts of the deletion of particular proteases are discussed, particular in relation to industrial applications.

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Section: Intracellular Proteasesmentioning

confidence: 99%

The ins and outs of Bacillus proteases: activities, functions and commercial significance

Harwood

Kikuchi

2021

FEMS Microbiology Reviews

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“…The Gram-negative E. coli represents a well-established and heavily used host for the production and purification of recombinant proteins. However, E. coli has some major drawbacks including the presence of endotoxins (LPS) or limitations in the secretion of proteins into the growth medium, which permits easier downstream processing (Lakowitz et al 2018 ; Terpe 2006 ). In contrast, Gram-positive bacteria lack an outer membrane, thereby omitting endotoxins and making protein secretion much more efficient.…”

Section: Production Of Recombinant Proteins Using P Megaterium : Intra- Und Extracellular Formation At G/l Scalementioning

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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“…Among thermophiles, thermophilic Bacillaceae possess several advantages for recombinant production of secreted thermostable enzymes. Most Bacillaceae strains are nonpathogenic, and more importantly, are particularly suited to secrete recombinant proteins [ 15 , 16 ]. Members of the Bacillus genus have been shown to secrete a large number of proteins into their environment, among them α-amylases and proteases, mainly through the general secretory pathway (Sec) which is the most commonly used secretion pathway in Gram-positive bacteria for biotechnological purposes [ 17 , 18 ].…”

Section: Introductionmentioning

confidence: 99%

Establishment of a functional system for recombinant production of secreted proteins at 50 °C in the thermophilic Bacillus methanolicus

et al. 2020

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Background: The suitability of bacteria as microbial cell factories is dependent on several factors such as price of feedstock, product range, production yield and ease of downstream processing. The facultative methylotroph Bacillus methanolicus is gaining interest as a thermophilic cell factory for production of value-added products from methanol. The aim of this study was to expand the capabilities of B. methanolicus as a microbial cell factory by establishing a system for secretion of recombinant proteins. Results: Native and heterologous signal peptides were tested for secretion of α-amylases and proteases, and we have established the use of the thermostable superfolder green fluorescent protein (sfGFP) as a valuable reporter protein in B. methanolicus. We demonstrated functional production and secretion of recombinant proteases, α-amylases and sfGFP in B. methanolicus MGA3 at 50 °C and showed that the choice of signal peptide for optimal secretion efficiency varies between proteins. In addition, we showed that heterologous production and secretion of α-amylase from Geobacillus stearothermophilus enables B. methanolicus to grow in minimal medium with starch as the sole carbon source. An in silico signal peptide library consisting of 169 predicted peptides from B. methanolicus was generated and will be useful for future studies, but was not experimentally investigated any further here. Conclusion: A functional system for recombinant production of secreted proteins at 50 °C has been established in the thermophilic B. methanolicus. In addition, an in silico signal peptide library has been generated, that together with the tools and knowledge presented in this work will be useful for further development of B. methanolicus as a host for recombinant protein production and secretion at 50 °C.

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Mini review: Recombinant production of tailored bio-pharmaceuticals in different Bacillus strains and future perspectives

Cited by 26 publications

References 164 publications

The ins and outs of Bacillus proteases: activities, functions and commercial significance

The ins and outs of Bacillus proteases: activities, functions and commercial significance

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

Establishment of a functional system for recombinant production of secreted proteins at 50 °C in the thermophilic Bacillus methanolicus

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