Effects of wet heat treatment on the germination of individual spores of Clostridium perfringens

G. Wang, D. Paredes-Sabja, M. R. Sarker, C. Green, P. Setlow, Y. Q. Li

Resultado de la investigación: Article

17 Citas (Scopus)

Resumen

Aim: To analyse the effect of wet heat treatment on nutrient and non-nutrient germination of individual spores of Clostridium perfringens. Methods and Results: Raman spectroscopy and differential interference contrast (DIC) microscopy were used to monitor the dynamic germination of individual untreated and wet heat-treated spores of Cl. perfringens with various germinants. When incubated in water at 90-100°C for 10-30 mmore than 90% of spores were inactivated but 50-80% retained their Ca2+-dipicolinic acid (CaDPA). The wet heat-treated spores that lost CaDPA exhibited extensive protein denaturation as seen in the 1640-1680 cm-1 (amide I) and 1230-1340 cm-1 (amide III) regions of Raman spectra, while spores that retained CaDPA showed partial protein denaturation. Wet heat-treated spores that retained CaDPA germinated with KCl or l-asparagine, but wet heat treatment increased values of Tlag, ΔTrelease and ΔTlys, during which spores initiated release of the majority of their CaDPA after mixing with germinant, released >90% of their CaDPA and completed the decrease in their DIC intensity because of cortex hydrolysis, respectively. Untreated Cl. perfringens spores lacking the essential cortex-lytic enzyme (CLE), SleC, exhibited longer Tlag and ΔTrelease values during KCl germination than wild-type spores and germinated poorly with CaDPA. Wet heat-treated wild-type spores germinating with CaDPA or dodecylamine exhibited increased Tlag, ΔTrelease and ΔTlys values, as did wet heat-treated sleC spores germinating with dodecylamine. Conclusions: (i) Some proteins important in Cl. perfringens spore germination are damaged by wet heat treatment; (ii) the CLE SleC or the serine protease CspB that activates SleC might be germination proteins damaged by wet heat; and (iii) the CaDPA release process seems likely to be damaged by wet heat. Significance and Impact of the Study: This study provides information on the germination of individual Cl. perfringens spores and improves the understanding of effects of wet heat treatment on spores.

Idioma originalEnglish
Páginas (desde-hasta)824-836
Número de páginas13
PublicaciónJournal of Applied Microbiology
Volumen113
N.º4
DOI
EstadoPublished - oct 2012

Huella dactilar

Clostridium perfringens
Germination
Spores
Hot Temperature
Protein Denaturation
Amides
Interference Microscopy
dipicolinic acid
Raman Spectrum Analysis
Asparagine
Serine Proteases
Enzymes

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Biotechnology
  • Medicine(all)

Citar esto

Wang, G. ; Paredes-Sabja, D. ; Sarker, M. R. ; Green, C. ; Setlow, P. ; Li, Y. Q. / Effects of wet heat treatment on the germination of individual spores of Clostridium perfringens. En: Journal of Applied Microbiology. 2012 ; Vol. 113, N.º 4. pp. 824-836.
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abstract = "Aim: To analyse the effect of wet heat treatment on nutrient and non-nutrient germination of individual spores of Clostridium perfringens. Methods and Results: Raman spectroscopy and differential interference contrast (DIC) microscopy were used to monitor the dynamic germination of individual untreated and wet heat-treated spores of Cl. perfringens with various germinants. When incubated in water at 90-100°C for 10-30 mmore than 90{\%} of spores were inactivated but 50-80{\%} retained their Ca2+-dipicolinic acid (CaDPA). The wet heat-treated spores that lost CaDPA exhibited extensive protein denaturation as seen in the 1640-1680 cm-1 (amide I) and 1230-1340 cm-1 (amide III) regions of Raman spectra, while spores that retained CaDPA showed partial protein denaturation. Wet heat-treated spores that retained CaDPA germinated with KCl or l-asparagine, but wet heat treatment increased values of Tlag, ΔTrelease and ΔTlys, during which spores initiated release of the majority of their CaDPA after mixing with germinant, released >90{\%} of their CaDPA and completed the decrease in their DIC intensity because of cortex hydrolysis, respectively. Untreated Cl. perfringens spores lacking the essential cortex-lytic enzyme (CLE), SleC, exhibited longer Tlag and ΔTrelease values during KCl germination than wild-type spores and germinated poorly with CaDPA. Wet heat-treated wild-type spores germinating with CaDPA or dodecylamine exhibited increased Tlag, ΔTrelease and ΔTlys values, as did wet heat-treated sleC spores germinating with dodecylamine. Conclusions: (i) Some proteins important in Cl. perfringens spore germination are damaged by wet heat treatment; (ii) the CLE SleC or the serine protease CspB that activates SleC might be germination proteins damaged by wet heat; and (iii) the CaDPA release process seems likely to be damaged by wet heat. Significance and Impact of the Study: This study provides information on the germination of individual Cl. perfringens spores and improves the understanding of effects of wet heat treatment on spores.",
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Effects of wet heat treatment on the germination of individual spores of Clostridium perfringens. / Wang, G.; Paredes-Sabja, D.; Sarker, M. R.; Green, C.; Setlow, P.; Li, Y. Q.

En: Journal of Applied Microbiology, Vol. 113, N.º 4, 10.2012, p. 824-836.

Resultado de la investigación: Article

TY - JOUR

T1 - Effects of wet heat treatment on the germination of individual spores of Clostridium perfringens

AU - Wang, G.

AU - Paredes-Sabja, D.

AU - Sarker, M. R.

AU - Green, C.

AU - Setlow, P.

AU - Li, Y. Q.

PY - 2012/10

Y1 - 2012/10

N2 - Aim: To analyse the effect of wet heat treatment on nutrient and non-nutrient germination of individual spores of Clostridium perfringens. Methods and Results: Raman spectroscopy and differential interference contrast (DIC) microscopy were used to monitor the dynamic germination of individual untreated and wet heat-treated spores of Cl. perfringens with various germinants. When incubated in water at 90-100°C for 10-30 mmore than 90% of spores were inactivated but 50-80% retained their Ca2+-dipicolinic acid (CaDPA). The wet heat-treated spores that lost CaDPA exhibited extensive protein denaturation as seen in the 1640-1680 cm-1 (amide I) and 1230-1340 cm-1 (amide III) regions of Raman spectra, while spores that retained CaDPA showed partial protein denaturation. Wet heat-treated spores that retained CaDPA germinated with KCl or l-asparagine, but wet heat treatment increased values of Tlag, ΔTrelease and ΔTlys, during which spores initiated release of the majority of their CaDPA after mixing with germinant, released >90% of their CaDPA and completed the decrease in their DIC intensity because of cortex hydrolysis, respectively. Untreated Cl. perfringens spores lacking the essential cortex-lytic enzyme (CLE), SleC, exhibited longer Tlag and ΔTrelease values during KCl germination than wild-type spores and germinated poorly with CaDPA. Wet heat-treated wild-type spores germinating with CaDPA or dodecylamine exhibited increased Tlag, ΔTrelease and ΔTlys values, as did wet heat-treated sleC spores germinating with dodecylamine. Conclusions: (i) Some proteins important in Cl. perfringens spore germination are damaged by wet heat treatment; (ii) the CLE SleC or the serine protease CspB that activates SleC might be germination proteins damaged by wet heat; and (iii) the CaDPA release process seems likely to be damaged by wet heat. Significance and Impact of the Study: This study provides information on the germination of individual Cl. perfringens spores and improves the understanding of effects of wet heat treatment on spores.

AB - Aim: To analyse the effect of wet heat treatment on nutrient and non-nutrient germination of individual spores of Clostridium perfringens. Methods and Results: Raman spectroscopy and differential interference contrast (DIC) microscopy were used to monitor the dynamic germination of individual untreated and wet heat-treated spores of Cl. perfringens with various germinants. When incubated in water at 90-100°C for 10-30 mmore than 90% of spores were inactivated but 50-80% retained their Ca2+-dipicolinic acid (CaDPA). The wet heat-treated spores that lost CaDPA exhibited extensive protein denaturation as seen in the 1640-1680 cm-1 (amide I) and 1230-1340 cm-1 (amide III) regions of Raman spectra, while spores that retained CaDPA showed partial protein denaturation. Wet heat-treated spores that retained CaDPA germinated with KCl or l-asparagine, but wet heat treatment increased values of Tlag, ΔTrelease and ΔTlys, during which spores initiated release of the majority of their CaDPA after mixing with germinant, released >90% of their CaDPA and completed the decrease in their DIC intensity because of cortex hydrolysis, respectively. Untreated Cl. perfringens spores lacking the essential cortex-lytic enzyme (CLE), SleC, exhibited longer Tlag and ΔTrelease values during KCl germination than wild-type spores and germinated poorly with CaDPA. Wet heat-treated wild-type spores germinating with CaDPA or dodecylamine exhibited increased Tlag, ΔTrelease and ΔTlys values, as did wet heat-treated sleC spores germinating with dodecylamine. Conclusions: (i) Some proteins important in Cl. perfringens spore germination are damaged by wet heat treatment; (ii) the CLE SleC or the serine protease CspB that activates SleC might be germination proteins damaged by wet heat; and (iii) the CaDPA release process seems likely to be damaged by wet heat. Significance and Impact of the Study: This study provides information on the germination of individual Cl. perfringens spores and improves the understanding of effects of wet heat treatment on spores.

KW - Clostridium perfringens

KW - Dipicolinic acid

KW - Spore germination

KW - Spores

KW - Wet heat inactivation

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U2 - 10.1111/j.1365-2672.2012.05387.x

DO - 10.1111/j.1365-2672.2012.05387.x

M3 - Article

C2 - 22776375

AN - SCOPUS:84866269878

VL - 113

SP - 824

EP - 836

JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

SN - 1364-5072

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