應用聚合酶鏈反應（PCR）技術鑑定分離自傳統家庭釀造液之醋酸菌（Acetobacter sp.）

Identification of Acetic Acid Bacteria Isolates from Home Made Fermentation Liquids by PCR

本研究根據醋酸菌之adb III序列(GenBank accession no.AB264314)，發展特異性PCR引子，ADH187/ADH500，此PCR引子成功的應用於醋酸菌菌株及非醋酸菌菌株之檢測。將此PCR引子應用於家庭自釀醋中分離之79株疑似醋酸菌株之檢測，其中41株經PCR引子初步確認為醋酸菌。以醋酸產生能力分析篩選出29株，進一步分析29株分離之醋酸菌之醋酸產量及酒精耐受度，發現分離株AAB305於含7%(v/v)乙醇之Yeast Glucose Manni tol/Mg 2+ (YGM/Mg2+medium中比其他分離株產生更高醋酸量，將來可進一步應用於工業上醋酸之生產。將醋酸產量高之分離株經16S rDNA序列分析確認皆為Acetobacter spp.。由結果顯示，此PCR方法與16S rDNA序列分析結果符合，將PCR方法應用於醋酸菌的篩選可節省大量人力及物力。

In this study, polymerase chain reaction primers based on adh III sequence (GenBank accession no. AB264314) were designed for identification of Acetobocter strains and acetic acid bacteria (AAB) isolates. This primer set termed ADH 187/ADH 500 was successfully used for identification of 41 AAB strains of 79 isolates directly from homemade vinegar fermentation. Furthermore, the acetate oxidation ability was used for analysis of acetate oxidation ability. The results showed that the AAB305 strain could produce higher acetate content than other AAB isolates in Yeast Glucose Mannitol/Mg(superscript 2+) (YGM/Mg(superscript 2+)) medium supplemented with 7% ethanol and could be used for industrial vinegar production. In addition, these isolates with high acetate oxidation ability were identified as Acetobacter sp. by the sequencing of the 16S rDNA PCR amplification products.