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

高淑真(Shu-Chen Kao) ； 廖文昌(Wayne-Chang Liao) ； 陳清惠(Ching-Hui Chen) ； 邱義源(Robin Yih-Yuan Chiou) ； 張育彰(Yu-Chang Chang)

《長庚科技學刊》 20期 (2014/06) Pp. 59-69

https://doi.org/10.6192/CGUST.2014.6.20.8

摘要

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。

關鍵字

蜂蜜 ； 芽孢桿菌 ； 核黃素

並列摘要

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＂.

並列關鍵字

honey ； Bacillus spp. ； riboflavin