Fluorescent In Situ Hybridization Analysis of Open Lactic Acid Fermentation of Kitchen Refuse Using rRNA-Targeted Oligonucleotide Probes(ENVIRONMENTAL BIOTECHNOLOGY)

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概要

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• Reproducible amounts of lactic acid accumulate in minced kitchen refuse under open conditions with intermittent pH neutralization [Sakai et al., Food Sci. Technol. Res., 6, 140 (2000)]. Here, we showed that such pH-controlled open fermentation of kitchen refuse reproducibly resulted a selective proliferation of a major lactic acid bacterial (LAB) species. In one experiment, the predominant microorganisms isolated during the early phase (6 h) were Gammaproteobacteria. In contrast, those that predominated during the late phase (48 h) were always Lactobacillus plantarum in three independent experiments. To further quantify the microbial community within open lactic acid fermentation, we performed fluorescent in situ hybridization (FISH) analysis targeting 16S (23S) rRNA. We designed two new group-specific DNA probes : LAC722 (L) was active for most LAB including the genera Lactobacillus, Pediococcus, Leuconostoc and Weisella, whereas Lplan477 was specific for L. plantarum and its related species. We then optimized sample preparation using lysozyme and hybridization conditions including temperature, as well as the formamide concentration and the salt concentration in the washing buffer. We succeeded in quantification of microorganisms in semi-solid, complex biological materials such as minced kitchen refuse by taking color microphotographs in modified RGB balance on pre-coated slides. FISH analysis of the fermentation of kitchen refuse indicated that control of the pH swing leads to domination by the LAB population in minced kitchen refuse under open conditions. We also confirmed that L. plantarum, which generates lactic acid in high quantities but with low optical activity, became the dominant microorganism in kitchen refuse during the late phase of open fermentation.

• 社団法人日本生物工学会の論文
• 2004-07-25

著者

• 酒井 謙二

  九大院生資環

• 酒井 謙二

  九州大学大学院農学研究院

• Fujii Akira

  Department of Cardiology, Ehime National Hospital

• Sakai Kenji

  Department of Plant Resources, Kyushu University

• MORI MASATSUGU

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• IWAMI YUKO

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• CHUKEATIROTE EKACHAI

  Department of Applied Chemistry, Faculty of Engineering, Oita University

• SHIRAI YOSHIHITO

  Department of Biochemical Engineering, Graduate School of Life Science and Systems Engineering, Kyus

• Moriguchi Mitsuaki

  Department Of Food And Bioscience Beppu University

• Iwami Yuko

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Sakai Kenji

  Department Of Physics Nagoya University

• Mori Masatsugu

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Shirai Y

  Department Of Biochemical Engineering And Science Kyushu Institute Of Technology

• Shirai Y

  Department Of Biochemical Engineering Graduate School Of Life Science And Systems Engineering Kyushu

• Shirai Yoshihito

  Department Of Biochemical Engineering And Science Faculty Of Computer Science And Systems Engineerin

• Sakai Kenji

  Department Of Environmental Chemistry And Engineering Faculty Of Engineering Oita University

• SAKAI Kenji

  Research Institute for Theoretical Physics Hiroshima University

• Chukeatirote Ekachai

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Chukeatirote Ekachai

  Department Of Applied Chemistry Faculty Of Engineering Oita University:department Of Biotechnology S

• Shirai Yoshihito

  Deparment Of Food Science And Technology Faculty Of Agriculture Kyoto University

• Sakai Kenji

  Department Of Drug Metabolism And Pharmacokinetics Nippon Boehringer Ingelheim Co. Ltd.

• Fujii Akira

  Department Of Applied Chemistry Faculty Of Engineering Oita University

• Sakai Kenji

  Department Of Applied Chemistry Faculty Of Engineering Oita University

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