Efficient secreted expression of natural intracellular β-galactosidase from Bacillus aryabhattai via non-classical protein secretion pathway in Bacillus subtilis

Duan, Xuguo; Luan, Shuyue

doi:10.1016/j.ijbiomac.2023.125758

Cited by 4 publications

(1 citation statement)

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“…Despite this handicap and also the high level of protease secreted by B. subtilis 168, secretion was high enough for Apr and SacB secretion tags to show green colour in the liquid culture when X-Gal was added (Supplementary Figure 8C). Nevertheless, we always recommend to test the non-classical protein secretion pathway of B. subtilis by using no tag, that can be much more efficient than the secretion tags as reported before and confirmed with the STK using galactosidase (Duan and Luan, 2023) (Supplementary figure 8C).…”

Section: Validation Of N-and C-terminal Assembliesmentioning

confidence: 60%

SubtiToolKit - a bioengineering kit forBacillus subtilisand Gram-positive bacteria

Caro-Astorga,

Rogan,

Malci

et al. 2024

Preprint

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SummaryBuilding DNA constructs of increasing complexity is key to synthetic biology. Golden Gate methods led to the creation of cloning toolkits - collections of modular standardized DNA parts hosted on hierarchic plasmids, developed for yeast, plants, Gram-negative bacteria, and human cells. However, Gram-positive bacteria have been neglected.Bacillus subtilisis a Gram-positive model organism and a workhorse in the bioindustry. Here, we present the SubtiToolKit, a high- efficiency cloning toolkit forB. subtilisand Gram-positive bacteria. Its design permits DNA constructs for transcriptional units, operons, knock-in and knock-out applications. It contains libraries of promoters, RBSs, fluorescent proteins, protein tags, terminators, genome integration parts, a no-leakage genetic device to control the expression of toxic products duringE. coliassembly, and a toolbox for industrially relevant strains ofGeobacillusandParageobacillusas an example of SubtiToolKit versatility for other Gram-positive bacteria and its future perspective as a reference toolkit.

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Section: Validation Of N-and C-terminal Assembliesmentioning

confidence: 60%