Phylogeny of γ-polyglutamic acid-producing Bacillus strains isolated from a fermented locust bean product manufactured in West Africa

Meerak, Jomkhwan; Yukphan, Pattaraporn; Miyashita, Mika; Sato, Hajime; Nakagawa, Yasuyoshi; Tahara, Yasutaka

doi:10.2323/jgam.54.159

Cited by 37 publications

(18 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the second step, ␥PGA-positive strains were selected. B. subtilis and B. amyloliquefaciens dominated the microbes found on the rice straw, although other Bacillus species, such as B. pumilus, B. atrophaeus, and B. licheniformis, produced ␥PGA under the conditions we employed (17).…”

Section: Resultsmentioning

confidence: 95%

Phylogenetic Analysis of Bacillus subtilis Strains Applicable to Natto (Fermented Soybean) Production

Kubo¹,

Rooney

Tsukakoshi

et al. 2011

Appl Environ Microbiol

106

View full text Add to dashboard Cite

Spore-forming Bacillus strains that produce extracellular poly-␥-glutamic acid were screened for their application to natto (fermented soybean food) fermentation. Among the 424 strains, including Bacillus subtilis and B. amyloliquefaciens, which we isolated from rice straw, 59 were capable of fermenting natto. Biotin auxotrophism was tightly linked to natto fermentation. A multilocus nucleotide sequence of six genes (rpoB, purH, gyrA, groEL, polC, and 16S rRNA) was used for phylogenetic analysis, and amplified fragment length polymorphism (AFLP) analysis was also conducted on the natto-fermenting strains. The ability to ferment natto was inferred from the two principal components of the AFLP banding pattern, and natto-fermenting strains formed a tight cluster within the B. subtilis subsp. subtilis group.The Bacillus subtilis species complex is a tight assemblage of related species that includes the laboratory strain B. subtilis subsp. subtilis 168, which has been used as a model organism for 32). B. subtilis subsp. subtilis BEST195, the starter strain used for natto (fermented soybean food) production, is thought to have been isolated about 100 years ago. Since then, it has been distributed to major commercial manufacturers (28). Before the isolation of the starter strain, dried rice straw had been used to initiate natto fermentation; rice straw is the natural habitat of the spore-forming B. subtilis strains that facilitate soybean fermentation. Recently, the entire genome information of the BEST195 strain was published (21). Completion of this project has facilitated the creation of a genomically directed starter cultivation program for natto. Natto starter cultivation targets genes regulating the metabolic pathways of secondary metabolites that affect flavor, sensitivity to bacteriophages, spore formation, and germination.The genome structure of B. subtilis subsp. subtilis BEST195 is very similar to that of the B. subtilis laboratory strain B. subtilis subsp. subtilis 168 (15, 21, 32), but it is not possible to produce natto with the latter (21), apparently due to mutations in the genes degQ and swrA, which regulate poly-␥-glutamic acid (␥PGA) synthesis (27). In contrast, BEST195 actively produces ␥PGA via the pgs operon (1, 2, 4), which provides a sticky texture to natto. However, ␥PGA production alone is not a predictor of the ability to ferment natto, as many ␥PGA-positive strains cannot be used for natto production. These observations have spurred great interest in the genetic basis of natto fermentation ability in Bacillus. Reports on the diversity of natto-fermenting strains are limited to elucidating the genetic background required for natto fermentation. To clarify the taxonomic positions of such strains within the genus Bacillus and to increase genetic sources for cultivating starters, we screened ␥PGA-positive strains from rice straw to determine their abilities to ferment natto.The commercial natto starter strain BEST195 requires biotin for growth. It is sensitive to specific bacteriophages (10, 25)...

show abstract

Section: Resultsmentioning

confidence: 95%

Phylogenetic Analysis of Bacillus subtilis Strains Applicable to Natto (Fermented Soybean) Production

Kubo¹,

Rooney

Tsukakoshi

et al. 2011

Appl Environ Microbiol

106

View full text Add to dashboard Cite

show abstract

“…B. subtilis and B. pumilus are capable of producing g-polyglutamic acid (PGA), and have been used in the processing of fermented soybean foods. 2 It is important to establish a rapid and simple method for identification of Bacillus spp. to assist in diagnosing diseases, anticipating public health hazards, recognizing warfare threats, and for monitoring possible contamination in stored foods and grains.…”

Section: Introductionmentioning

confidence: 99%

Lipid fingerprinting of Bacillus spp. using online MALDI-TOF mass spectrometry

et al. 2012

View full text Add to dashboard Cite

Rapid identification of Bacillus spores has attracted great interest in the medical and forensic communities due to the highly pathogenic nature of certain Bacillus species and its potential application in bioweapons. In this study, we evaluated the feasibility of rapid and reliable identification of several Bacillus spp. (B. subtilis, B. cereus, B. megaterium, B. pumilus, and B. sphaericus) using online MALDI-TOF mass spectrometry combined with lipid profiling. The bacterial cultures in liquid medium were extracted with methanol and chloroform, and the extracts were aerosolized into droplets and immediately analyzed with an online MALDI-TOF mass spectrometer. Results showed that phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and diglycosyldiacylglycerol (DGDG) were present in the mass spectra, as alkali metal adducts. The unique lipid profiles obtained from these Bacillus spp. allowed us to differentiate between them. The reliability and reproducibility of the rapid identification method for Bacillus were tested under different experimental conditions. An alternative online chemotaxonomic method for bacterial identification is demonstrated.

show abstract

“…2). At the close of the analysis B. amyloliquefaciens strains GH58 and GH28 (Meerak et al, 2008) were identified as being almost identical, i.e. with 99% identity.…”

Section: Resultsmentioning

confidence: 95%

Identification a Novel Raw-Starch-Degrading-α-Amylase from a Tropical Marine Bacterium

Nurachman¹

2010

American Journal of Biochemistry and Biotechnology

View full text Add to dashboard Cite

Problem statement: Bacteria from the surface of the tropical marine hard coral Acropora sp. were screened for producing raw-starch-degrading-α-amylase. Approach: Based on its 16s rDNA sequence, a bacterium that produced the highest amylolitic activity was identified as Bacillus amyloliquifaciens ABBD. The bacterial isolate secreted a α-amylase extracellularly and then the enzyme was partially purified by ammonium sulfate precipitation followed by anion exchange chromatography. Results: Electrophoresis results both SDS-PAGE and native PAGE suggested that the enzyme was a heterodimeric protein (97 kDa) consisting of 45 and 55 kDa subunits. The α-amylase had an optimum pH of 7.0 and temperature of 60°C. More than 80% activity of the enzyme was retained under high salt conditions (up to 20% NaCl). The enzyme remained stable at 50°C for 1 h. Starch hydrolysis by the enzyme at 70°C yielded oligosaccharides (G2-G4) and at room temperature yielded glucose/maltose (G1 and G2). Conclusion: The B. amyloliquifaciens ABBD α-amylase was capable of degrading various raw starch granules from corn, rice, cassava and sago at room temperature.

show abstract

Phylogeny of γ-polyglutamic acid-producing Bacillus strains isolated from a fermented locust bean product manufactured in West Africa

Cited by 37 publications

References 24 publications

Phylogenetic Analysis of Bacillus subtilis Strains Applicable to Natto (Fermented Soybean) Production

Phylogenetic Analysis of Bacillus subtilis Strains Applicable to Natto (Fermented Soybean) Production

Lipid fingerprinting of Bacillus spp. using online MALDI-TOF mass spectrometry

Identification a Novel Raw-Starch-Degrading-α-Amylase from a Tropical Marine Bacterium

Contact Info

Product

Resources

About

Phylogeny of γ-polyglutamic acid-producing Bacillus strains isolated from a fermented locust bean product manufactured in West Africa

Cited by 37 publications

References 24 publications

Phylogenetic Analysis of Bacillus subtilis Strains Applicable to Natto (Fermented Soybean) Production

Phylogenetic Analysis of Bacillus subtilis Strains Applicable to Natto (Fermented Soybean) Production

Lipid fingerprinting of Bacillus spp. using online MALDI-TOF mass spectrometry

Identification a Novel Raw-Starch-Degrading-&alpha;-Amylase from a Tropical Marine Bacterium

Contact Info

Product

Resources

About

Identification a Novel Raw-Starch-Degrading-α-Amylase from a Tropical Marine Bacterium