Effects of manganese (II) on Bacillus licheniformis ATCC 9945A physiology and γ-poly(glutamic acid) formation

Cromwick, Anne-Marie; Gross, Richard A.

doi:10.1016/0141-8130(95)98153-p

Cited by 71 publications

(63 citation statements)

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“…The ratio between L-and D-glutamate in Bacillus licheniformis c-PGA also depends on the concentration of Co 2+ and Zn 2+ . Other researchers have also confirmed the dependence of the enantiomeric composition of c-PGA on Mn 2+ in both B. licheniformis and B. subtilis (Cromwick & Gross, 1995a;Wu et al, 2006).…”

Section: Mechanism Of Synthesis Of C-pgamentioning

confidence: 57%

Poly-γ-glutamic acid: production, properties and applications

et al. 2015

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Section: Mechanism Of Synthesis Of C-pgamentioning

confidence: 57%

Poly-γ-glutamic acid: production, properties and applications

et al. 2015

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“…This is a considerably shorter incubation period than that used by others, which varied between 48 and 96 h (2,3,5), but this probably reflects the simplicity and low concentrations of medium components used here. Although depolymerization of PGA ensues as the bacterium enters stationary phase, this is a slow process (2,4,6), and electrophoresis of precipitated PGA showed that there was no substantial degradation until incubation beyond 30 h (Fig. 2).…”

Section: Strains Of Bacillus Licheniformis Andmentioning

confidence: 99%

Regulation of Polyglutamic Acid Synthesis by Glutamate in Bacillus licheniformis and Bacillus subtilis

Kambourova

Tangney

Priest

2001

Appl Environ Microbiol

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The synthesis of polyglutamic acid (PGA) was repressed by exogenous glutamate in strains of Bacillus licheniformis but not in strains of Bacillus subtilis, indicating a clear difference in the regulation of synthesis of capsular slime in these two species. Although extracellular ␥-glutamyltranspeptidase (GGT) activity was always present in PGA-producing cultures of B. licheniformis under various growth conditions, there was no correlation between the quantity of PGA and enzyme activity. Moreover, the synthesis of PGA in the absence of detectable GGT activity in B. subtilis S317 indicated that this enzyme was not involved in PGA biosynthesis in this bacterium. Glutamate repression of PGA biosynthesis may offer a simple means of preventing unwanted slime production in industrial fermentations using B. licheniformis. Strains of Bacillus licheniformis andBacillus subtilis may synthesize a water-soluble, viscous slime material containing Dand L-glutamic acid residues. This polyglutamic acid (PGA) is polymerized via amide linkages between the ␣-amino and ␥-carboxylic groups of the amino acid residues. PGA is the principal component of "Itohiki-natto," a traditional Japanese food prepared from steamed soybean by the biological action of PGA-producing strains of B. subtilis, which are generally referred to as "Bacillus natto" or B. subtilis (natto) (17). PGA has other biotechnological applications in cosmetics, medicines, and foods. However, the synthesis of even small amounts of PGA can be a problem in the fermentation industry, most notably in the production of extracellular enzymes from bacilli, where PGA accumulation causes increased viscosity of the fermentation broth, reduced enzyme yield, uncontrollable foaming, and complications in product recovery. The unpredictable nature of PGA synthesis is particularly troublesome.Cultural conditions affecting PGA biosynthesis have been studied using various poorly identified strains of B. licheniformis and B. subtilis in complex, ill-defined media, and these studies have resulted in conflicting conclusions. In general, strains have been classified into two categories: (i) those that require exogenous L-glutamate for PGA synthesis, for example, B. subtilis strains IFO 3335 (6) and F-2-01 and B. licheniformis 9945 (16) (although Birrer et al. reported that strain 9945 does not require exogenous glutamate for PGA synthesis [2]), and (ii) strains that do not require exogenous glutamate (often described as de novo synthesis), such as B. licheniformis A35 (3) and B. subtilis TAM-4 (10).The mechanisms of PGA biosynthesis in B. licheniformis and B. subtilis have been elusive until recently. A membranous synthetase complex from B. licheniformis which catalyzes the activation, racemization, and polymerization of L-glutamate into exclusively poly-D-glutamate has been partially characterized (5), but B. subtilis (natto) has been studied more extensively in this context. A gene originally thought to code for ␥-glutamyltranspeptidase (GGT) (8) but later referred to as a PGA "stimulating ...

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“…PGA was first discovered as a component of the capsule of Bacillus anthracis (19) and Bacillus mesentericus (18) and was isolated from the culture medium of Bacillus subtilis (7). Since then, a number of bacteria producing PGA, including B. subtilis (8,17,21,22), Bacillus licheniformis (9,42), and Bacillus megaterium (13,41), have been reported. PGA is a main constituent of the sticky material in natto, a Japanese traditional food made from soybeans that have been steamed and then fermented by B. subtilis (14).…”

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Characterization of the Bacillus subtilis ywsC Gene, Involved in γ-Polyglutamic Acid Production

2002

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The genes required for ␥-polyglutamic acid (PGA) production were cloned from Bacillus subtilis IFO16449, a strain isolated from fermented soybeans. There were four open reading frames in the cloned 4.2-kb DNA fragment, and they were almost identical to those in the ywsC and ywtABC genes of B. subtlis 168. Northern blot analysis showed that the four genes constitute an operon. Three genes, ywsC, ywtA, and ywtB, were disrupted to determine which gene plays a central role in PGA biosynthesis. No PGA was produced in ⌬ywsC and ⌬ywtA strains, indicating that both of these genes are essential for PGA production. To clarify the function of the YwsC protein, histidine-tagged YwsC (YwsC-His) was produced in the ⌬ywsC strain and purified from the lysozyme-treated lysate of the transformant by Ni-nitrilotriacetic acid affinity chromatography. Western blot analysis revealed that the YwsC-His protein consists of two subunits, the 44-kDa and 33-kDa proteins, which are encoded by in-phase overlapping in the ywsC gene.14 C-labeled PGA was synthesized by the purified proteins from L- Some Bacillus strains produce ␥-polyglutamic acid (PGA), an amino acid polymer that consists of only D-glutamic acid or D-and L-glutamic acid polymerized through ␥-glutamyl bonds, as a capsular or an extracellular viscous material (6). PGA was first discovered as a component of the capsule of Bacillus anthracis (19) and Bacillus mesentericus (18) and was isolated from the culture medium of Bacillus subtilis (7). Since then, a number of bacteria producing PGA, including B. subtilis (8,17,21,22), Bacillus licheniformis (9, 42), and Bacillus megaterium (13, 41), have been reported. PGA is a main constituent of the sticky material in natto, a Japanese traditional food made from soybeans that have been steamed and then fermented by B. subtilis (14).Concerning PGA biosynthesis, Makino et al. reported cloning of three genes, capBCA, responsible for capsular PGA biosynthesis from B. anthracis, and the gene products occurred together as membrane-associated proteins in the Escherichia coli transformant (26,27,43). The complete genome sequence of B. subtilis 168, in which ywsC and ywtAB were found to be highly homologous to the capBCA genes of B. anthracis, has been made available in databases (31). Recently, pgsBCA genes for PGA biosynthesis were also cloned from B. subtilis IFO3336, and their sequences were found to be the same as those of the ywsC and ywtAB genes of B. subtilis 168 (2). These three genes seem to be involved in PGA production; however, little is known about the function of each gene product in PGA biosynthesis.In this paper, we describe the cloning and gene disruption of the ywsC and ywtAB genes, which are responsible for PGA production in B. subtilis IFO16449, a strain isolated from natto, and we also describe the characterization of the YwsC 44-kDa and 33-kDa proteins, which catalyze the biosynthesis of PGA from L-glutamate, a crucial enzyme in PGA production. MATERIALS AND METHODSBacterial strains, plasmids, and media. Bacillus subtilis IF...

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Effects of manganese (II) on Bacillus licheniformis ATCC 9945A physiology and γ-poly(glutamic acid) formation

Cited by 71 publications

References 11 publications

Poly-γ-glutamic acid: production, properties and applications

Poly-γ-glutamic acid: production, properties and applications

Regulation of Polyglutamic Acid Synthesis by Glutamate in Bacillus licheniformis and Bacillus subtilis

Characterization of the Bacillus subtilis ywsC Gene, Involved in γ-Polyglutamic Acid Production

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