蜂蜜中芽孢桿菌分離、鑑定與酵素活性分析

Isolation, Identification and Enzyme Profile of Bacillus Strains Isolated from Honey

本研究收集46件台灣與各國的蜂蜜樣本，其中33件分離出芽孢桿菌，分離率71.7%。分離的102株芽孢桿菌菌種以B.cereus group(50.0%)及B.subtilis(30.4%)為主，其他尚包括B.amyloliquefaciens、B.circulans、B.fusiformis、B.gibsonii、B.indicus、B.megaterium及B.pumilus等9種。大部分樣本分離出1或2種芽孢桿菌菌種，但一件「龍眼花蜜」樣本可分離出5種芽孢桿菌。挑選5株芽孢桿菌使用API ZYM套組，進行19種酵素活性分析，結果顯示不同蜂蜜樣本分離的B.cereus group（HC5、HC11及HT64等三株）間的酵素活性差異大，但均有高的leucine aminopeptidase酵素活性，B.indicus HT68對於β-galactosidase有高的酵素活性，B.gibsonii HT71無較明顯的測試酵素活性。此外，以聚合鏈反應方法檢測分離株的核黃素基因，在102株測試株中有34株帶有核黃素基因，檢出率33.3%，這些菌株主要為B.cereus group及B.subtilis。本研究初步將不同國家及地區所生產的蜂蜜進行芽孢桿菌分離、鑑定及酵素活性分析，未來將探討具有應用價值潛力的芽孢桿菌。

Of the 46 honey samples collected from different countries, 33 (71.7%) had the Bacillus spp. isolated. Following the identification tests, the major isolates were identified as B. cereus group (50.0%) and B. subtilis (30.4%). Others including B. amyloliquefaciens, B. circulans, B. fusiformis, B. gibsonii, B. indicus, B. megaterium and B. pumilus. In addition, most of the samples had less than two Bacillus spp. isolated, with only one “longan honey” sample resulted in five Bacillus species being identified. Among these, five strains were selected and subjected to analysis by API-ZYM kit for studying the possible presence activities of 19 enzymes. Results demonstrated the properties of different enzymes varied between strains for the B. cereus group (HC5, HC11 and HT64) as isolated from different honey samples. However, all the strains studied showed a high leucine aminopeptidase activity, of which, higher level of β-galactosidase activity in B. indicus HT68 was observed. Yet, the B. gibsonii HT71 strain revealed no significant difference in enzyme activity. Moreover, the detection of riboflavin (rib) genes by PCR showed a positive yield in 34 out of 102 isolates, with a detection rate of 33.3% which are mainly belong to B. cereus group and B. subtilis. This study tried to separate and identify the Bacillus strains from the honey samples produced in different countries, and analyzed the enzyme activity. Further research will try to explore the potential for the application of Bacillus.