利用分子標誌區分楊桃鹽醃發酵過程中酵母菌之菌相

Abstract

本研究之目的在於探討甜味種與酸味種楊桃鹽醃發酵過程酵母菌菌相之變化，並對其揮發性化合物進行分析研究。 由發酵甜味種楊桃醃漬液及發酵酸楊桃醃漬液F及L（分別取自彰化縣花壇及雲林縣林內地區）進行酵母菌菌相區分，分別取30、60、90天之楊桃醃漬液於DRBC培養基進行培養，培養3至5天後，挑選外觀型態不同之酵母菌單一菌落共37株，接種至PDA培養基進行純化保存，再利用引子ITS1及ITS4進行PCR將分離純化之酵母菌株rRNA的ITS1至ITS2 DNA片段擴增，並進行基因定序，所得序列與Genbank資料庫比對分析，結果做為酵母菌分類之用。 取發酵30、60、90天之楊桃醃漬液共9個樣品進行揮發性化合物分析，再利用GC/MS以SPME的方法進行分析，而得知甜味種與酸味種楊桃醃漬液揮發性成分各有異同。 發酵醃漬30天之各樣品及樣品F之各時期皆能分離出Debaryomyces hansnii var. fabryi。甜楊桃醃漬液分類出Candida humilis，Candida tropicalis，Pichia anomala，Hanseninaspora uvarum四株酵母菌與前人研究相同。酸楊桃醃漬液樣品皆未分類出與前人研究相同之菌株。 樣品F發酵90天與樣品L發酵60天所分離出之菌株分類比對結果相同。揮發性化合物分析結果，除樣品F發酵30天分析出之Methyl benzoate外，其餘皆符合前人研究之結果。綜合以上風味描述，不論甜味種或酸味種楊桃經發酵後之醃漬液為具有果香、酒香、甜香、花香、清香等綜合香氣特性。

The aim of study is found out that the change of yeast phases during fermentation between the salt pickling fermented tart and sweet carambola juice. The salt pickling fermented sweet and tart (F&L) carambola, cultured the samples 3-5 days on DRBC media after fermented for 30, 60, 90 days, choose and get 37 varieties the single colony from the plates, Clone on PDA for pure culture. Employ primers ITS1 & ITS4 to PCR, the products use to genome sequencing. The sequences are assay by Genbank. The parted of anlysis volatile compounds, use GC/MS by the SPME method. Present different result to the volatile compounds, between different variety sweet and tart caranbola. All samples after 30 days could separate Debaryomyces hansnii var. faryi, the sample of sweet carambola could identity Candida humilis, Candida tropicalis, Pichia anomala, Hanseninaspora uvarum, the same with the other study. The samples of tart carambola can’t identity the same yeast with other studies. But the different area tart carambola gets the same yeast. Tart carambola pickling brine F fermented after 90 days and tart carambola pickling brine L fermented after 60 days had the same yeasts of class with stain. The volatility assay from tart carambola pickling brine F fermented after 30 days had methyl benzoate was different with the other study. Integrate the aroma describe, however, sweet or tart carambola pickling brine had fruit aroma, alcohol aroma, sweet aroma, flower aroma and faint scent.