Presence of Acylated Homoserine Lactones (APresence of Acylated Homoserine Lactones (AHLs) andHLs) and

AHL-Producing Bacteria in Meat and PotentialAHL-Producing Bacteria in Meat and PotentialRole of AHL in Spoilage of MeatRole of AHL in Spoilage of Meat

Jesper Bartholin Bruhn,1* Allan Beck Christensen,2 Lars Ravn FlodgJesper Bartholin Bruhn,1* Allan Beck Christensen,2 Lars Ravn Flodgaard,1aard,1

Kristian Fog Nielsen,3 Thomas Ostenfeld Larsen,3 Michael Givskov,Kristian Fog Nielsen,3 Thomas Ostenfeld Larsen,3 Michael Givskov,22

and Lone Gram1and Lone Gram1

Received 26 September 2003/Accepted 8 March 2004Received 26 September 2003/Accepted 8 March 2004

學生學生 ::吳俊昌吳俊昌

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, July 2004, p. 4293–4302 Vol. 70, No. 7

IntroductionIntroduction

Enterobacteriaceae Enterobacteriaceae 腸道菌腸道菌 種類 例種類 例 ::

Hafnia alvei Hafnia alvei 718718腐肉中最多 腐肉中最多

Serratia spp.Serratia spp.次多次多

Serratia proteamaculansSerratia proteamaculans B5a B5a產產 proteaseprotease 使牛奶使牛奶 coagulationcoagulation

IntroductionIntroduction

Monitor bacteriaMonitor bacteria

Agrobacterium tumefaciens Agrobacterium tumefaciens NT1(pZLR4)NT1(pZLR4)

AHL-reporter plasmidAHL-reporter plasmid

偵測有偵測有 oxo,6-14Coxo,6-14C

Chromobacterium violaceum Chromobacterium violaceum CV026CV026

偵測無偵測無 oxo,4Coxo,4C

IntroductionIntroduction

AHLAHL 有不同的有不同的 structure structure 例例 ::

N-3-oxo-hexanoyl homoserine lactone(OHHL)N-3-oxo-hexanoyl homoserine lactone(OHHL)腐肉中佔最多腐肉中佔最多比沒有比沒有 oxooxo 的利用率的利用率 ,, 效能及持久度高效能及持久度高

N-hexanoyl homoserine lactone(HHL)N-hexanoyl homoserine lactone(HHL)腐肉中次多腐肉中次多

N-octanoyl homoserine lactone(OHL)N-octanoyl homoserine lactone(OHL)

菌數濃度菌數濃度

分離菌種分離菌種

主要的主要的 AHLAHL 構型構型1.Marker

2.H. alvei 718

3.4 week 再萃取

4.3 week (fat)

5.2 week (fat)

6.1 week (low fat)

7. 當天 (fat)

利用 ESI+-MS 再度證實

AHLAHL 影響影響 phenotypephenotype

1.marker

2.718

3.718

4.718 halⅠ mutant

5.718 halⅠ mutant

6.718 halⅠ mutant + OHHL

7.718 halⅠ mutant + OHHL

8.marker

20kDa

Bacteria communicationBacteria communication

SummarySummary

Hafnia alvei Hafnia alvei 718 718 為主要為主要 以以 N-3-oxo-hexanoyl homoserine lactoneN-3-oxo-hexanoyl homoserine lactone

(OHHL)(OHHL) 為主要構型為主要構型718718 halⅠ halⅠ mutantmutant 影響影響 phenotypephenotypeBacteria communication(718 and B5a)Bacteria communication(718 and B5a)