Targeting the vanillic acid decarboxylase gene for Alicyclobacillus acidoterrestris quantification and guaiacol assessment in apple juices using real time PCR.
Targeting the vanillic acid decarboxylase gene for Alicyclobacillus acidoterrestris quantification and guaiacol assessment in apple juices using real time PCR.
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DOI：
10.1016/j.ijfoodmicro.2020.109006
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发表时间：
2020-12
影响因子：
5.4
通讯作者：
Zhouli Wang;T. Yue;Yahong Yuan;Yanchen Zhang;Zhenpeng Gao;Rui Cai
中科院分区：
文献类型：
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作者：
Zhouli Wang;T. Yue;Yahong Yuan;Yanchen Zhang;Zhenpeng Gao;Rui Cai
近年来，由于其在巴氏灭菌果汁的变质中所起的作用，引起了人们的广泛关注，其特征是形成愈创木酚。以往的研究表明并非所有的aliicyclobacillusspp。都能生产愈创木酚。本研究旨在鉴定愈创木酚生物合成过程中涉及的香草酸脱羧酶基因的可能差异，并建立产生愈创木酚的酸环杆菌的特异性检测方法。琼脂糖凝胶电泳结果显示，产愈创木酚的藻环杆菌中均存在部分vdcci基因，而除产愈创木酚的菌株ma外，非产愈创木酚的菌株中不存在vdcci基因。fastidiosusDSM 17978。在vdccgene序列的基础上，找到了一个特异的引物对。acidoterrestriswas设计;建立SYBR Green I实时荧光定量PCR，直接定量a的含量。检测限为2.6 × 101CFU/mL。采用所建立的实时PCR系统检测ta。用气相色谱-质谱法分析了36份人工污染苹果汁样品的抑酸作用和愈创木酚的产生。采用Gompertz模型来描述a和a之间的关系。酸抑制细胞浓度和愈创木酚含量，r2值为0.854。这项工作为传统的愈创木酚定量方法提供了一种替代方法。对a的早期识别非常有用。酸碱抑制果汁中的污染。
Alicyclobacillus spp. has recently received much attention due to its implication in the spoilage of pasteurized fruit juices, which is characterized by the formation of guaiacol. Previous researches indicate that not allAlicyclobacillusspp. are able to produce guaiacol. The aim of this study was to identify possible differences in the vanillic acid decarboxylase gene involved in guaiacol biosynthesis and then develop specific detection methods for guaiacol producingAlicyclobacillus. Agarose gel electrophoresis results showed that the partialvdcCgene was present in all the guaiacol producingAlicyclobacillus, but absent in non-guaicaol producing strains apart fromA. fastidiosusDSM 17978. On the basis of thevdcCgene sequence, a primer pair specific toA. acidoterrestriswas designed; then a SYBR Green I real time PCR was established for the direct quantification ofA. acidoterrestrisin apple juice, and the detection limit was 2.6 × 101CFU/mL. The developed real time PCR system was used to detectA. acidoterrestrisin 36 artificially contaminated apple juice samples and guaiacol production in the sample was also analyzed by GC–MS. The Gompertz model was employed to describe the relationship betweenA. acidoterrestriscell concentration and guaiacol content, and the value ofR2was 0.854. This work provides an alternative to conventional methods of guaiacol quantification andA. acidoterrestrisdetection and could be very useful for the early recognition ofA. acidoterrestriscontamination in fruit juices.