蜂粮中芽孢杆菌的分离鉴定及其耐酸耐高糖特性的分析
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摘要
为丰富人工发酵蜂花粉生产蜂粮的菌种资源,从天然蜂粮中筛选芽孢杆菌。通过平板涂布法从中蜂蜂粮样品中分离产芽孢的细菌,利用16S rRNA基因同源比对及系统发育分析对其进行初步鉴定,并分析其耐酸耐高糖特性。共分离到16株隶属于Bacillus tequilensis、Bacillus aerophilus、Fictibacillus nanhaiensis、Bacillus sonorensis、Lysinibacillus xylanilyticus、Lysinibacillus fusiformis、Bacillus invictus、Bacillus methylotrophicus的芽孢杆菌。其中10株菌能在低酸和高糖条件下正常生长,菌株Br7、Br9和Br11的芽孢生成量较少,有潜力应用于人工发酵蜂花粉制备蜂粮。
Bacillus strains were isolated from natural bee bread to enrich microbes producing bee bread from the fermentation of bee pollen. Strains were isolated based on the homologous alignmentusing spread plate methodandpreliminaryidentified using phylogenetic analysis of the 16 S rRNA genes. Theirtolerance to acidand glucosewas also discussed. Sixteen spore-forming bacteria were totally isolated from bee bread of Apissinensis and identified as the genus of Bacillus tequilensis,Bacillus aerophilus,Fictibacillusnanhaiensis,Bacillus sonorensis,Lysinibacillusxylanily-ticus,Lysinibacillusfusiformis,Bacillus invictus and Bacillus methylotrophicus,respectively.However,only ten Bacillus strains could grow vigorously at low pH(4.0) with high glucose concentration(30%). Fewer spores were produced by the strain Br7,Br9 and Br11,which were suggested as the potential strains used for artificial fermentation of bee pollen.
Bacillus strains were isolated from natural bee bread to enrich microbes producing bee bread from the fermentation of bee pollen. Strains were isolated based on the homologous alignmentusing spread plate methodandpreliminaryidentified using phylogenetic analysis of the 16 S rRNA genes. Theirtolerance to acidand glucosewas also discussed. Sixteen spore-forming bacteria were totally isolated from bee bread of Apissinensis and identified as the genus of Bacillus tequilensis,Bacillus aerophilus,Fictibacillusnanhaiensis,Bacillus sonorensis,Lysinibacillusxylanily-ticus,Lysinibacillusfusiformis,Bacillus invictus and Bacillus methylotrophicus,respectively.However,only ten Bacillus strains could grow vigorously at low pH(4.0) with high glucose concentration(30%). Fewer spores were produced by the strain Br7,Br9 and Br11,which were suggested as the potential strains used for artificial fermentation of bee pollen.
引文
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[15]GHOSH J,LARSSON P,SINGH B,et al.Sporulation in mycobacteria[J].Proceedings of the National Academy of Sciences,2009,106(26):10781-10786.
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[2]NAGAI T,NAGASHIMA T,NAGAI T,et al.Preparation and functional properties of extracts from bee bread[J].Food/Nahrung,2004,48(3):226-229.
[3]O’TOOLE C,RAW A.Bees of the World[M].Blandford Press,1991.
[4]HUANG Wencheng.Microorganism is important for nutrition and health of swarm[J].Apiculture of China,2010,61(4):54-55.(in Chinese)
[5]BRINDZA J,GROF J,BACIGALOVA K,et al.Pollen microbial colonization and food safety[J].Acta Chimica Slovaca,2010,3(1):95-102.
[6]HAYDEN C.Microbiology of pollen and bee bread:The genus Bacillus[J].Apidologie,1979,10(3):269-275.
[7]ISIDOROVA V A,ISIDOROVAB A G,SCZCZEPANIAKA L,et al.Gas chromatographic-mass spectrometric investigation of the chemical composition of beebread[J].Food Chemistry,2009,115:1056-1063.
[8]GILLIAM M,PREST D B,LORENZ B J.Microbiology of pollen and bee bread:taxonomy and enzymology of molds[J].Apidologie,1989,20:53-68.
[9]SU Songkun,CHEN Shenglu,YU Xuping,et al.Isolation and identification of bacteria from pollen and bee bread[J].Journal of Zhejiang University(Agric.&Life Sci.),2001,27(6):627-630.(in Chinese)
[10]FOOTE H L.Possible use of microorganisms in synthetic bee bread production[J].Amer Bee J,1957,97:476-478.
[11]HAYDEN C.Microbiology of pollen and bee bread:The Genus Bacillus[J].Apidologie,1979,10(3):269-275.
[12]JI Baozhong,LIU Shuwen,XU Lina,et al.Advances on the bee bread research[J].Chinese Agricultural Science Bulletin,2006,22(8):165-169.(in Chinese)
[13]WEI Juan,HE Dongxu,LI Guohong,et al.Isolation and identification of endophytic bacteria in Paris polyphylla Smith var.yunnanensis(Franch)and phylogenetic analysis[J].Journal of Food Science and Biotechnology,2015,34(2):165-169.(in Chinese)
[14]HAN Huili,CAI Yujie,GUAN Zhengbing,et al.Screening and identification and study on antimicrobial activity of bacterial isolates from honey[J].Journal of Food Science and Biotechnology,2013,32(2):148-154.(in Chinese)
[15]GHOSH J,LARSSON P,SINGH B,et al.Sporulation in mycobacteria[J].Proceedings of the National Academy of Sciences,2009,106(26):10781-10786.
[16]WANG L,LIN Y M.Spore detection in aerobic granules by different dipicolinic acid releasing methods[J].Bioresource Technology,2007,98:3164-3167.