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Alkyl hydroperoxide reductase enhances the growth of Leuconostoc mesenteroides lactic acid bacteria at low temperatures.

Goto S, Kawamoto J, Sato SB, Iki T, Watanabe I, Kudo K, Esaki N, Kurihara T - AMB Express (2015)

Bottom Line: NH04 grew rapidly at low temperatures within the shelf-life period and resulted in heavy financial losses.We introduced an expression plasmid containing ahpC into NBRC3832, which grows slower than NH04 at 10°C, and found that expression of AhpC enhanced growth.These results demonstrated that AhpC, which likely increases anti-oxidative capacity of LAB, plays an important role in their rapid growth at low temperatures.

View Article: PubMed Central - PubMed

Affiliation: Product Development Laboratory, NH Foods Ltd., Chikusei, Ibaraki 308-0042 Japan.

ABSTRACT
Lactic acid bacteria (LAB) can cause deterioration of food quality even at low temperatures. In this study, we investigated the cold-adaptation mechanism of a novel food spoilage LAB, Leuconostoc mesenteroides NH04 (NH04). L. mesenteroides was isolated from several spoiled cooked meat products at a high frequency in our factories. NH04 grew rapidly at low temperatures within the shelf-life period and resulted in heavy financial losses. NH04 grew more rapidly than related strains such as Leuconostoc mesenteroides NBRC3832 (NBRC3832) at 10°C. Proteome analysis of NH04 demonstrated that this strain produces a homolog of alkyl hydroperoxide reductase--AhpC--the expression of which can be induced at low temperatures. The expression level of AhpC in NH04 was approximately 6-fold higher than that in NBRC3832, which was grown under the same conditions. Although AhpC is known to have an anti-oxidative role in various bacteria by catalyzing the reduction of alkyl hydroperoxide and hydrogen peroxide, the involvement of AhpC in cold adaptation of food spoilage bacteria was unclear. We introduced an expression plasmid containing ahpC into NBRC3832, which grows slower than NH04 at 10°C, and found that expression of AhpC enhanced growth. These results demonstrated that AhpC, which likely increases anti-oxidative capacity of LAB, plays an important role in their rapid growth at low temperatures.

No MeSH data available.


Related in: MedlinePlus

Effects ofahpCoverexpression on the growth of NBRC3832. (a) Production of AhpC from NH04 in NBRC3832. Soluble proteins from NBRC3832 harboring a control plasmid (lane 1) and an ahpC overexpression plasmid (lane 2) were analyzed by SDS-PAGE. Arrowhead indicates the band corresponding to AhpC. Lane M represents the molecular weight marker. (b) Growth of NH04 (square) and NBRC3832 (triangle) containing a control plasmid (filled symbol) and an ahpC overexpression plasmid (open symbol) at 10°C. Each growth curve was plotted using data obtained from three experiments.
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Fig3: Effects ofahpCoverexpression on the growth of NBRC3832. (a) Production of AhpC from NH04 in NBRC3832. Soluble proteins from NBRC3832 harboring a control plasmid (lane 1) and an ahpC overexpression plasmid (lane 2) were analyzed by SDS-PAGE. Arrowhead indicates the band corresponding to AhpC. Lane M represents the molecular weight marker. (b) Growth of NH04 (square) and NBRC3832 (triangle) containing a control plasmid (filled symbol) and an ahpC overexpression plasmid (open symbol) at 10°C. Each growth curve was plotted using data obtained from three experiments.

Mentions: To examine the effects of high-level expression of AhpC on the growth of LAB at low temperatures, we constructed an overexpression vector for ahpC (pGKahpC) and introduced it into NBRC3832, which grows slower than NH04. The level of AhpC in NBRC3832 harboring pGKahpC was approximately 6.5 ± 1.6-fold higher than that in the cells harboring a control plasmid, pGK::nucMCS (Figure 3a). NBRC3832 overexpressing ahpC grew more rapidly than the cells containing a control plasmid at 10°C (Figure 3b). The doubling time of NBRC3832 harboring pGKahpC was 15.4 ± 0.8 h, whereas that of the strain harboring pGK::nucMCS was 17.6 ± 1.1 h. The difference between the growth rates of NBRC3832 harboring pGKahpC and pGK::nucMCS was statistically significant (p < 0.05, n = 3). In contrast to the case of NBRC3832, the growth rate of NH04 was not affected by introduction of pGKahpC (Figure 3b). The doubling times of NH04 harboring pGKahpC and pGK::nucMCS were both 9.8 ± 0.3 h at 10°C. At 25°C, introduction of pGKahpC did not significantly affect the growth rate of NBRC3832 and NH04 (data not shown). The doubling times of NBRC3832 harboring pGKahpC and pGK::nucMCS were 3.0 ± 0.2 h and 3.2 ± 0.1 h, respectively, at 25°C, which were not significantly different from each other (p < 0.05, n = 3). The doubling times of NH04 harboring pGKahpC and pGK::nucMCS were both 2.0 ± 0.1 h at 25°C.Figure 3


Alkyl hydroperoxide reductase enhances the growth of Leuconostoc mesenteroides lactic acid bacteria at low temperatures.

Goto S, Kawamoto J, Sato SB, Iki T, Watanabe I, Kudo K, Esaki N, Kurihara T - AMB Express (2015)

Effects ofahpCoverexpression on the growth of NBRC3832. (a) Production of AhpC from NH04 in NBRC3832. Soluble proteins from NBRC3832 harboring a control plasmid (lane 1) and an ahpC overexpression plasmid (lane 2) were analyzed by SDS-PAGE. Arrowhead indicates the band corresponding to AhpC. Lane M represents the molecular weight marker. (b) Growth of NH04 (square) and NBRC3832 (triangle) containing a control plasmid (filled symbol) and an ahpC overexpression plasmid (open symbol) at 10°C. Each growth curve was plotted using data obtained from three experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
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Fig3: Effects ofahpCoverexpression on the growth of NBRC3832. (a) Production of AhpC from NH04 in NBRC3832. Soluble proteins from NBRC3832 harboring a control plasmid (lane 1) and an ahpC overexpression plasmid (lane 2) were analyzed by SDS-PAGE. Arrowhead indicates the band corresponding to AhpC. Lane M represents the molecular weight marker. (b) Growth of NH04 (square) and NBRC3832 (triangle) containing a control plasmid (filled symbol) and an ahpC overexpression plasmid (open symbol) at 10°C. Each growth curve was plotted using data obtained from three experiments.
Mentions: To examine the effects of high-level expression of AhpC on the growth of LAB at low temperatures, we constructed an overexpression vector for ahpC (pGKahpC) and introduced it into NBRC3832, which grows slower than NH04. The level of AhpC in NBRC3832 harboring pGKahpC was approximately 6.5 ± 1.6-fold higher than that in the cells harboring a control plasmid, pGK::nucMCS (Figure 3a). NBRC3832 overexpressing ahpC grew more rapidly than the cells containing a control plasmid at 10°C (Figure 3b). The doubling time of NBRC3832 harboring pGKahpC was 15.4 ± 0.8 h, whereas that of the strain harboring pGK::nucMCS was 17.6 ± 1.1 h. The difference between the growth rates of NBRC3832 harboring pGKahpC and pGK::nucMCS was statistically significant (p < 0.05, n = 3). In contrast to the case of NBRC3832, the growth rate of NH04 was not affected by introduction of pGKahpC (Figure 3b). The doubling times of NH04 harboring pGKahpC and pGK::nucMCS were both 9.8 ± 0.3 h at 10°C. At 25°C, introduction of pGKahpC did not significantly affect the growth rate of NBRC3832 and NH04 (data not shown). The doubling times of NBRC3832 harboring pGKahpC and pGK::nucMCS were 3.0 ± 0.2 h and 3.2 ± 0.1 h, respectively, at 25°C, which were not significantly different from each other (p < 0.05, n = 3). The doubling times of NH04 harboring pGKahpC and pGK::nucMCS were both 2.0 ± 0.1 h at 25°C.Figure 3

Bottom Line: NH04 grew rapidly at low temperatures within the shelf-life period and resulted in heavy financial losses.We introduced an expression plasmid containing ahpC into NBRC3832, which grows slower than NH04 at 10°C, and found that expression of AhpC enhanced growth.These results demonstrated that AhpC, which likely increases anti-oxidative capacity of LAB, plays an important role in their rapid growth at low temperatures.

View Article: PubMed Central - PubMed

Affiliation: Product Development Laboratory, NH Foods Ltd., Chikusei, Ibaraki 308-0042 Japan.

ABSTRACT
Lactic acid bacteria (LAB) can cause deterioration of food quality even at low temperatures. In this study, we investigated the cold-adaptation mechanism of a novel food spoilage LAB, Leuconostoc mesenteroides NH04 (NH04). L. mesenteroides was isolated from several spoiled cooked meat products at a high frequency in our factories. NH04 grew rapidly at low temperatures within the shelf-life period and resulted in heavy financial losses. NH04 grew more rapidly than related strains such as Leuconostoc mesenteroides NBRC3832 (NBRC3832) at 10°C. Proteome analysis of NH04 demonstrated that this strain produces a homolog of alkyl hydroperoxide reductase--AhpC--the expression of which can be induced at low temperatures. The expression level of AhpC in NH04 was approximately 6-fold higher than that in NBRC3832, which was grown under the same conditions. Although AhpC is known to have an anti-oxidative role in various bacteria by catalyzing the reduction of alkyl hydroperoxide and hydrogen peroxide, the involvement of AhpC in cold adaptation of food spoilage bacteria was unclear. We introduced an expression plasmid containing ahpC into NBRC3832, which grows slower than NH04 at 10°C, and found that expression of AhpC enhanced growth. These results demonstrated that AhpC, which likely increases anti-oxidative capacity of LAB, plays an important role in their rapid growth at low temperatures.

No MeSH data available.


Related in: MedlinePlus