Improved catalytic and antifungal activities of<i>Bacillus thuringiensis</i>cells with surface display of Chi9602ΔSP

Tang, Min; Sun, Xiaowen; Zhang, Shan; Wan, Juan; Li, Lin; Hóng, Ní

doi:10.1111/jam.13333

Cited by 14 publications

(5 citation statements)

References 34 publications

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“…Localization of enzymes on cell membrane is a general method used for increasing the protein's activity and stability. In Bacillus thuringiensis , chitinase was re‐located on the cell membrane with anchoring motifs and resulted in increased chitinase activity and stability (Tang et al, 2017). In E. coli , xylanase activity was increased by the re‐location on the cell membrane with anchoring motifs (Chen et al, 2012).…”

Section: Resultsmentioning

confidence: 99%

Engineering Corynebacterium glutamicum for synthesis of poly(3‐hydroxybutyrate) from lignocellulose biomass

Jin

Huang

et al. 2022

Biotech & Bioengineering

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Lignocellulose is the only feasible carbohydrates feedstock for commercial scale and carbon neutral production of poly(3‐hydroxybutyrate) (PHB) biopolymer by its great abundance and availability. Microbial cell factories for fermentative PHB synthesis are highly restricted by the growth suppression of inhibitors from lignocellulose pretreatment. This study targeted a potential PHB‐producing cell factory Corynebacterium glutamicum owing to its strong inhibitors tolerance. A systematic metabolic engineering was conducted starting with the stable PHB synthesis pathway construction from glucose and xylose, followed by the enhancement of PHB synthesis on PHA synthase activity and stability, cell morphology modification, and growth factors regulation. The relocation of the PHA synthase on the cell membrane guided by secrete signal peptides and cell membrane display motifs increased the PHB content by 2.4 folds. Excessive nitrogen preferentially promoted the PHB synthesis capacity and resulted in the PHB content increased by 13.3 folds. Modification of the genes responsible for cell division changed the cell morphology but the cell size was not enlarged to a PHB accumulation favorable environment. The metabolic engineering of C. glutamicum resulted in a high fermentative production of PHB using wheat straw as feedstock. This study provided an important microbial cell factory choice for PHB production using lignocellulose feedstock.

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Section: Resultsmentioning

confidence: 99%

Engineering Corynebacterium glutamicum for synthesis of poly(3‐hydroxybutyrate) from lignocellulose biomass

Jin

Huang

et al. 2022

Biotech & Bioengineering

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“…In recent years, the surface display technology has been widely used in baculovirus, E. coli, yeast and mammalian cells. For examples, the surface display system constructed by the expression of outer membrane porin SmoA using Synechococcus elongatus can be used as a continuous bioproduct platform (Fedeson and Ducat 2016), the surface display systems constructed based on Bacillus thuringiensis (Bt) strain BMB171 using two N-terminal domains of autolysin as anchoring motifs can be used to improve catalytic and antibacterial activity of chitinase Chi9602DSP (Tang et al 2017), and the surface expression system VSV-Ebola based on vesicular stomatitis virus can be used to screen antibodies and vaccines against Ebola viruses (Khurana et al 2016). However, studies on cell display technology are mainly focused on bacteria and fungi, there are also some applications on insect cells.…”

Section: Discussionmentioning

confidence: 99%

Establishment of insect cell lines expressing green fluorescent protein on cell surface based on AcMNPV GP64 membrane fusion characteristic

Chen

et al. 2017

Cytotechnology

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Displaying a protein on the surface of cells has been provided a very successful strategy to function research of exogenous proteins. Based on the membrane fusion characteristic of Autographa californica multiple nucleopolyhedrovirus envelope protein GP64, we amplified and cloned N-terminal signal peptide and C-terminal transmembrane domain as well as cytoplasmic tail domain of gp64 gene into vector pIZ/V5-His with multi-cloning sites to construct the cell surface expression vector pIZ/V5-gp64. To verify that the vector can be used to express proteins on the membrane of insect cells, a recombinant plasmid pIZ/V5-gp64-GFP was constructed by introducing the PCR amplified green fluorescent protein (GFP) gene and transfected into insect cell lines Sf9 and H5. The transected cells were screened with zeocin and cell cloning. PCR verification results showed that the GFP gene was successfully integrated into these cells. Green fluorescence in Sf9-GFP and H5-GFP cells was observed by using confocal laser scanning microscopy and immunofluorescence detection indicated that GFP protein was located on the cell membrane. Western blot results showed that a fusion protein GP64-GFP of about 40 kDa was expressed on the membrane of Sf9-GFP and H5-GFP cells. The expression system constructed in this paper can be used for localization and continuous expression of exogenous proteins on insect cell membrane.

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“…The identical anchoring motif was utilized to develop a chitinolytic whole-cell B. thuringiensis biocatalyst. (Tang et al 2017 ). This biocatalyst displayed enhanced catalytic and antifungal activities.…”

Section: Molecular Tools Developed For Increased Production Of Recomb...mentioning

confidence: 99%

Bacillus and Streptomyces spp. as hosts for production of industrially relevant enzymes

Vojnovic,

Aleksic,

Ilic-Tomic

et al. 2024

Appl Microbiol Biotechnol

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The application of enzymes is expanding across diverse industries due to their nontoxic and biodegradable characteristics. Another advantage is their cost-effectiveness, reflected in reduced processing time, water, and energy consumption. Although Gram-positive bacteria, Bacillus, and Streptomyces spp. are successfully used for production of industrially relevant enzymes, they still lag far behind Escherichia coli as hosts for recombinant protein production. Generally, proteins secreted by Bacillus and Streptomyces hosts are released into the culture medium; their native conformation is preserved and easier recovery process enabled. Given the resilience of both hosts in harsh environmental conditions and their spore-forming capability, a deeper understanding and broader use of Bacillus and Streptomyces as expression hosts could significantly enhance the robustness of industrial bioprocesses. This mini-review aims to compare two expression hosts, emphasizing their specific advantages in industrial surroundings such are chemical, detergent, textile, food, animal feed, leather, and paper industries. The homologous sources, heterologous hosts, and molecular tools used for the production of recombinant proteins in these hosts are discussed. The potential to use both hosts as biocatalysts is also evaluated. Undoubtedly, Bacillus and Streptomyces spp. as production hosts possess the potential to take on a more substantial role, providing superior (bio-based) process robustness and flexibility. Key points • Bacillus and Streptomyces spp. as robust hosts for enzyme production. • Industrially relevant enzyme groups for production in alternative hosts highlighted. • Molecular biology techniques are enabling easier utilization of both hosts.

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Improved catalytic and antifungal activities ofBacillus thuringiensiscells with surface display of Chi9602ΔSP

Abstract: Results demonstrated that the bacterial cell display system can be used to express biocontrol agents on the cell surface of B. thuringiensis and provides an attractive method for inhibiting phytopathogenic fungi.

Cited by 14 publications

References 34 publications

Engineering Corynebacterium glutamicum for synthesis of poly(3‐hydroxybutyrate) from lignocellulose biomass

Engineering Corynebacterium glutamicum for synthesis of poly(3‐hydroxybutyrate) from lignocellulose biomass

Establishment of insect cell lines expressing green fluorescent protein on cell surface based on AcMNPV GP64 membrane fusion characteristic

Bacillus and Streptomyces spp. as hosts for production of industrially relevant enzymes

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