scholarly journals Advances in Enhanced Menaquinone-7 Production From Bacillus subtilis

2021 ◽  
Vol 9 ◽  
Author(s):  
Chaoyong Liao ◽  
Hammed Ayansola ◽  
Yanbo Ma ◽  
Koichi Ito ◽  
Yuming Guo ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Chemical Synthesis ◽  
Biosynthetic Pathway ◽  
Gene Editing ◽  
Vitamin K2 ◽  
Future Studies ◽  
Biofilm Reactors ◽  
Nutraceutical Compounds ◽  
Activation System ◽  
Crispr Activation

The production of nutraceutical compounds through biosynthetic approaches has received considerable attention in recent years. For example, Menaquinone-7 (MK-7), a sub-type of Vitamin K2, biosynthesized from Bacillus subtilis (B. subtilis), proved to be more efficiently produced than the conventional chemical synthesis techniques. This is possible due to the development of B. subtilis as a chassis cell during the biosynthesis stages. Hence, it is imperative to provide insights on the B. subtilis membrane permeability modifications, biofilm reactors, and fermentation optimization as advanced techniques relevant to MK-7 production. Although the traditional gene-editing method of homologous recombination improves the biosynthetic pathway, CRISPR-Cas9 could potentially resolve the drawbacks of traditional genome editing techniques. For these reasons, future studies should explore the applications of CRISPRi (CRISPR interference) and CRISPRa (CRISPR activation) system gene-editing tools in the MK-7 anabolism pathway.

scholarly journals Genome Sequence of Bacillus subtilis natto VK161, a Novel Strain That Produces Vitamin K2

2019 ◽  
Vol 8 (35) ◽  
Author(s):  
Dylan Parks ◽  
In-Hwa Chung ◽  
In kyu Lee ◽  
Eui-Joong Kim ◽  
Sarbjeet Niraula ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Vitamin K2 ◽  
Department Of Energy ◽  
Rrna Genes ◽  
Trna Genes ◽  
Protein Coding ◽  
Content Type ◽  
Bacillus Subtilis Natto ◽  
High Production ◽  
Energy Joint Genome Institute

Bacillus subtilis strain natto VK161 was selected for its high production of vitamin K2. Its genome was sequenced and annotated in the Department of Energy-Joint Genome Institute (DOE-JGI) annotation pipeline. It resulted in a chromosome of 4,073,396 bp, which is composed of 4,332 protein-coding genes, 23 rRNA genes, and 77 tRNA genes.

scholarly journals Ratio Between Lactobacillus Plantarum and Acetobacter Pomorum on the Surface of Drosophila Melanogaster Adult Flies Depends on Cuticle Melanisation

2021 ◽  
Author(s):  
Vladislav Mokeev ◽  
Yiwen Wang ◽  
Nicole Gehring ◽  
Bernard Moussian
Keyword(s):  
Drosophila Melanogaster ◽  
Lactobacillus Plantarum ◽  
Relative Abundance ◽  
Gene Editing ◽  
Fruit Fly ◽  
Wild Type ◽  
Specific Primers ◽  
Future Studies ◽  
Rdna Sequencing ◽  
Simple Protocol

Abstract Objectives As in most organisms, the surface of the fruit fly Drosophila melanogaster is associated with bacteria. In order to study the genetic parameters of this association, we developed a simple protocol for surface bacteria isolation and quantification. Results On wild-type flies maintained in the laboratory, we identified two persistently culturable species as Lactobacillus plantarum and Acetobacter pomorum by 16S rDNA sequencing. For quantification, we showered single flies for DNA extraction avoiding the rectum to prevent contamination from the gut. Using specific primers for quantitative PCR analyses, we determined the relative abundance of these two species in surface wash samples. Repeatedly, we found 20% more L. plantarum than A. pomorum . To tentatively study the importance of the cuticle for the interaction of the surface with these bacteria, applying Crispr/Cas9 gene editing in the initial wild-type flies, we generated flies mutant for the ebony gene needed for cuticle melanisation and determined the L. plantarum to A. pomorum ratio on these flies. We found that the relative abundance of L. plantarum increased substantially on ebony flies. We conclude that the cuticle chemistry is crucial for surface bacteria composition. This finding may inspire future studies on cuticle-microbiome interactions.

APPLICATION OF CRISPR ACTIVATION (CRISPRa) SYSTEM TO ZEBRAFISH (Danio rerio) GENES

2019 ◽  
Vol 5 (2) ◽  
Author(s):  
Rizka Rahmana Putri
Keyword(s):  
Danio Rerio ◽  
Mrna Level ◽  
Gene Activation ◽  
Chain Termination ◽  
Mrna Levels ◽  
Activation System ◽  
Dna Chain ◽  
Object Of Study ◽  
Crispr Activation

CRISPR activation system is part of the function of CRSPR/Cas9 which is used to manipulate certain genes by activating these genes. CRISPR activation utilizes dCas9 (dead-Cas9) or Cas9 which is deactivated as DNA scissors, so that the use of Cas9 for gene activation does not cause targeted DNA chain termination. This research that uses this CRISPR activation is applied to the zebrafish gene (Danio rerio) aims to determine the mRNA level of the targeted gene. In this study, the ASCL1a and BCL6a genes from zebrafish were targeted as the object of study. The results showed that genes from zebrafish that were targeted had an increase in mRNA levels after being activated using CRISPR activation system. Keywords: CRISPR activation system (CRISPRa), zebrafish (Danio rerio), dCas9, ASCL1a gene, BCL6a gene

scholarly journals A Polysaccharide Deacetylase Gene (pdaA) Is Required for Germination and for Production of Muramic δ-Lactam Residues in the Spore Cortex of Bacillus subtilis

2002 ◽  
Vol 184 (21) ◽  
pp. 6007-6015 ◽  
Author(s):  
Tatsuya Fukushima ◽  
Hiroki Yamamoto ◽  
Abdelmadjid Atrih ◽  
Simon J. Foster ◽  
Junichi Sekiguchi
Keyword(s):  
Bacillus Subtilis ◽  
Phase Contrast ◽  
Biosynthetic Pathway ◽  
Wild Type Strain ◽  
Dipicolinic Acid ◽  
Northern Hybridization ◽  
Predicted Amino Acid Sequence ◽  
Wild Type ◽  
Contrast Microscopy ◽  
Acid Release

ABSTRACT The predicted amino acid sequence of Bacillus subtilis yfjS (renamed pdaA) exhibits high similarity to those of several polysaccharide deacetylases. β-Galactosidase fusion experiments and results of Northern hybridization with sporulation sigma mutants indicated that the pdaA gene is transcribed by EσG RNA polymerase. pdaA-deficient spores were bright by phase-contrast microscopy, and the spores were induced to germination on the addition of l-alanine. Germination-associated spore darkening, a slow and partial decrease in absorbance, and slightly lower dipicolinic acid release compared with that by the wild-type strain were observed. In particular, the release of hexosamine-containing materials was lacking in the pdaA mutant. Muropeptide analysis indicated that the pdaA-deficient spores completely lacked muramic δ-lactam. A pdaA-gfp fusion protein constructed in strain 168 and pdaA-deficient strains indicated that the protein is localized in B. subtilis spores. The biosynthetic pathway of muramic δ-lactam is discussed.

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