title: Involvement of Colonizing Bacillus Isolates in Glucovanillin Hydrolysis during the Curing of Vanilla planifolia Andrews.
abstract: Vanilla beans were analyzed using biochemical methods, which revealed that glucovanillin disperses from the inner part to the outer part of the vanilla bean during the curing process and is simultaneously hydrolyzed by β-d-glucosidase. Enzymatic hydrolysis was found to occur on the surface of the vanilla beans. Transcripts of the β-d-glucosidase gene (bgl) of colonizing microorganisms were detected. The results directly indicate that colonizing microorganisms are involved in glucovanillin hydrolysis. Phylogenetic analysis based on 16S rRNA gene sequences showed that the colonizing microorganisms mainly belonged to the Bacillus genus. bgl was detected in all the isolates and presented clustering similar to that of the isolate taxonomy. Furthermore, inoculation of green fluorescent protein-tagged isolates showed that the Bacillus isolates can colonize vanilla beans. Glucovanillin was metabolized as the sole source of carbon in a culture of the isolates within 24 h. These isolates presented unique glucovanillin degradation capabilities. Vanillin was the major volatile compound in the culture. Other compounds, such as α-cubebene, β-pinene, and guaiacol, were detected in some isolate cultures. Colonizing Bacillus isolates were found to hydrolyze glucovanillin in culture, indirectly demonstrating the involvement of colonizing Bacillus isolates in glucovanillin hydrolysis during the vanilla curing process. Based on these results, we conclude that colonizing Bacillus isolates produce β-d-glucosidase, which mediates glucovanillin hydrolysis and influences flavor formation.
authors.affiliation: Spice and Beverage Research Institute, CATAS, Wanning, China College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture, Wanning, China.
authors.forename: Yonggan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Spice and Beverage Research Institute, CATAS, Wanning, China Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture, Wanning, China xiaogu4117@163.com.
authors.forename: Fenglin
authors.initials: F
authors.lastname: Gu
authors.affiliation: Agricultural Product Processing Research Institute, CATAS, Zhanjiang, China.
authors.forename: Jihua
authors.initials: J
authors.lastname: Li
authors.affiliation: Spice and Beverage Research Institute, CATAS, Wanning, China Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture, Wanning, China.
authors.forename: Shuzhen
authors.initials: S
authors.lastname: He
authors.affiliation: Spice and Beverage Research Institute, CATAS, Wanning, China Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture, Wanning, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Xu
authors.affiliation: Spice and Beverage Research Institute, CATAS, Wanning, China Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops, Ministry of Agriculture, Wanning, China.
authors.forename: Yiming
authors.initials: Y
authors.lastname: Fang
mesh_terms: D001407:Bacillus / Q000201:enzymology / Q000378:metabolism*; D001547:Benzaldehydes / Q000378:metabolism*; D016000:Cluster Analysis; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D031669:Vanilla / Q000737:chemistry* / Q000382:microbiology*; D001617:beta-Glucosidase / Q000378:metabolism*
chemical_list: D001547:Benzaldehydes; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; C100058:vanillin; D001617:beta-Glucosidase; C443341:glucovanillin