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通过氯霉素乙酰转移酶基因的随机整合监测发酵蔬菜中肠膜明串珠菌DRC发酵剂

Monitoring of Leuconostoc mesenteroides DRC starter in fermented vegetable by random integration of chloramphenicol acetyltransferase gene.

作者信息

Eom Hyun-Ju, Park Joong Min, Seo Min Jae, Kim Myoung-Dong, Han Nam Soo

机构信息

Department of Food Science and Technology, Research Center for Bioresource and Health, Chungbuk National University, Cheongju, 361-763, Korea.

出版信息

J Ind Microbiol Biotechnol. 2008 Sep;35(9):953-9. doi: 10.1007/s10295-008-0369-y. Epub 2008 May 24.

Abstract

In 2004, Leuconostoc mesenteroides DRC was first used as a starter culture for achieving higher organoleptic effects in Korean kimchi manufacture. For a better understanding of starter growth in a mixed culture system, and for predicting starter predominance in kimchi, a monitoring system for the starter was established. The chloramphenicol resistance marker gene (cat) was randomly integrated into chromosomal DNA of L. mesenteroides DRC using a viral transposon and transposase. The DRC mutant, tDRC2, had a similar growth pattern to the host strain, with no major alteration in phenotypic characteristics. The mutant strain was inoculated into real kimchi, and monitoring of the starter population was successfully achieved. The overall predominance of Leuconostoc in kimchi inoculated with DRC followed the general growth pattern of this genus during kimchi fermentation. Our results also demonstrate the competitive ability of the DRC starter against Leuconostoc from natural flora, maintaining its predominance above 88% during the whole fermentation period. Based on this experiment, the random gene integration method using a transposon was shown to be of utility in transferring any commercial starter into a selectable and monitorable strain for simulation purposes.

摘要

2004年,肠膜明串珠菌DRC首次被用作发酵剂,以在韩国泡菜制作中获得更高的感官效果。为了更好地理解混合培养系统中发酵剂的生长情况,并预测泡菜中发酵剂的优势地位,建立了一个发酵剂监测系统。使用病毒转座子和转座酶将氯霉素抗性标记基因(cat)随机整合到肠膜明串珠菌DRC的染色体DNA中。DRC突变体tDRC2具有与宿主菌株相似的生长模式,其表型特征没有重大改变。将突变菌株接种到真正的泡菜中,成功实现了对发酵剂群体的监测。接种DRC的泡菜中肠膜明串珠菌的总体优势地位遵循该属在泡菜发酵过程中的一般生长模式。我们的结果还证明了DRC发酵剂对来自天然菌群的肠膜明串珠菌的竞争能力,在整个发酵期内其优势地位保持在88%以上。基于该实验,使用转座子的随机基因整合方法被证明可用于将任何商业发酵剂转化为可选择和可监测的菌株,用于模拟目的。

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