该文分析榨菜腌制过程中与菜块表面变红和产膜现象相关微生物的种类和来源。通过分离鉴定变红或产膜相关的微生物,用平板水解圈法检测其产酶活性,采用Illumina Miseq高通量测序平台分析青菜头原料表面的微生物种类。结果表明,引起变红的菌株有4株,共3种,包括嗜吡啶红球菌(Rhodococcus pyridinivorans)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、地衣芽孢杆菌(Bacillus licheniformis)。引起产膜的菌株有13株,共7种,包括解淀粉芽孢杆菌 (Bacillus amyloliquefaciens)、贝莱斯芽孢杆菌 (Bacillus velezensis)、阿耶波多氏芽孢杆菌 (Bacillus aryabhattai)、枯草芽孢杆菌(Bacillus subtilis)、地衣芽胞杆菌(Bacillus licheniformis)、蜡状芽孢杆菌(Bacillus cereus)和表皮葡萄球菌(Staphylococcus epidermidis)。其中嗜吡啶红球菌和表皮葡萄球菌有可能来源于原料。因此,该研究可为榨菜变红和产膜现象的防控提供参考。
This study focused on the microorganisms related to the surface turned red or biofilm-forming of Zhacai during the pickling process. The samples that the surface turned red or biofilm-forming Zhacai were collected for the isolation and purification related microorganisms. Specific verification experiments were conducted to confirm their relationship with above phenomenon. Marker gene 16S rDNA and plate hydrolysis zone were used for the strain characterization and testing their enzyme-producing capability. Illumina Miseq sequencing technology can help to find out where the relevant microorganism come from. In this study, 4 strains were determined to be associated with Zhacai surface turning red phenomenon which including 3 species: Rhodococcus pyridinivorans, Bacillus amyloliquefaciens and Bacillus licheniformis. Moreover, 13 strains were showed to be related to the biofilm-forming phenomenon which belonging to 7 species: Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus aryabhattai, Bacillus subtilis, Bacillus licheniformis, Bacillus cereus and Staphylococcus epidermidis. Further study indicated that Rhodococcus pyridinivorans and Staphylococcus epidermidis were probably originated from raw mustard tuber material. The results of this study provided basic data for the prevention and controlling of microorganism that resulting in Zhacai surface turning red and biofilm-forming.
[1] 庞杰,向珣.涪陵榨菜的加工及生产现状[J].长江蔬菜, 1999(2): 42-44.
[2] 苏扬,张聪,王朝辉.榨菜加工工业及其发展战略的研究[J].食品科技,2010,35(4):114-119.
[3] 贺云川,周斌全,刘德君.涪陵榨菜传统工艺概述[J].食品与发酵科技, 2013,49(4):57-60.
[4] 邓维琴,伍亚龙,陈功,等.多代发酵泡菜母水微生物多样性及其理化指标动态研究[J].食品与发酵工业, 2018,44(8):49-56.
[5] XU Nan,TAN Guangcai,WANG Hongyuan,et al.Effect of biochar additions to soil on nitrogen leaching, microbial biomass and bacterial community structure[J].European Journal of Soil Biology,2016,74:1-8.
[6] 何志刚,李维新,梁璋成,等.优良降酸酿酒酵母的分离和鉴定[J].中国食品学报,2013,13(5):187-193.
[7] 王珍珍,沙如意,蔡成岗,等.树莓酵素中耐高渗酵母菌的分离鉴定及生长特性研究[J].食品工业科技,2017, 38(8):178-182;188.
[8] 吴丹,陈健初,叶兴乾,等.榨菜腐败微生物的分离、鉴定及生物学特性研究[J].浙江大学学报(农业与生命科学版),2009,35(2):135-140.
[9] YANG Jixia,CAO Jialu,XU Haiyan,et al.Bacterial diversity and community structure in Chongqing radish paocai brines revealed using PacBio SMRT sequencing technology[J].Journal of the Science of Food and Agriculture,2018, 98(9):3 234-3 245.
[10] CAPORASO J G,KUCZYNSKI J,STOMBAUGH J,et al.QIIME allows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336.
[11] EDGAR R C.Search and clustering orders of magnitude faster than BLAST[J].Bioinformatics,2010,26 (19):2 460-2 461.
[12] QUAST C,PRUESSE E,YILMAZ P,et al.The SILVA ribosomal RNA gene database project: Improved data processing and web-based tools[J].Nucleic Acids Research,2013,41(D1):D590-D596.
[13] YILMAZ P,PARFREY L W,YARZA P,et al. The SILVA and & quot;all-species living tree project (LTP) & quot; taxonomic frameworks[J].Nucleic Acids Research,2014,42(D1):D643-D648.
[14] LU Yingying,CHEN Jing,ZHENG Junyuan,et al.Mucosal adherent bacterial dysbiosis in patients with colorectal adenomas[J].Scientific reports,2016,6:26 337.
[15] 程池,刘光全,李金霞,等.55株芽孢杆菌16S rRNA基因序列测定与系统发育学分析[J].食品与发酵工业, 2006,32(10):20-24.
[16] STACKEBRANDT E,GOEBEL B M.Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology[J]. International Journal of Systematic Bacteriology,1994,44(4): 842-849.
[17] WANG L T,LEE F L,TAI C J,et al.Bacillus velezensis is a later heterotypic synonym of Bacillus amyloliquefaciens[J].International Journal of Systematic and Evolutionary Microbiology,2008,58(3):671-675.
[18] YOON J H,KANG S S,CHO Y G,et al.Rhodococcus pyridinivorans sp. nov. a pyridine-degrading bacterium[J]. International Journal of Systematic and Evolutionary Microbiology,2000,50(6):2 173-2 180.
[19] 郑晓冬,王友永.用红球菌生产类胡萝卜素的研究[J].浙江大学学报(农业与生命科学版),2000,26(5):516-520.
[20] TAKAICHI S,MAOKA T,AKIMOTO N,et al.Carotenoids in a corynebacterineae, Gordonia terrae AIST-1: Carotenoid glucosyl mycoloyl esters[J].Bioscience, Biotechnology and Biochemistry,2008,72(10):2 615-2 622.
[21] 刘振宇,曹阳,邓德法,等.解淀粉芽孢杆菌生物膜特征与生防活性[C].中国植物病理学会2012年学术年会论文集,2012:362.
[22] 蔡高磊,张凡,欧阳友香,等.贝莱斯芽孢杆菌(Bacillus velezensis)研究进展[J].北方园艺,2018(12):162-167.
[23] HONG B H,JOE M M,SELVAKUMAR G,et al.Influence of salinity variations on exocellular polysaccharide production, biofilm formation and flocculation in halotolerant bacteria[J].Journal of Environmental Biology,2017, 38(4):657-664.
[24] 刘伟杰,刘聪,蒋继宏.枯草芽孢杆菌形成生物被膜的研究进展[J].微生物学报,2014,54(9):977-983.
[25] RANDRIANJATOVO-GBALOU I,ROUQUETTE P,LEFEBVRE D,et al.In situ analysis of Bacillus licheniformis biofilms: Amyloid-like polymers and eDNA are involved in the adherence and aggregation of the extracellular matrix[J].Journal of Applied Microbiology,2017,122(5):1 262-1 274.
[26] MAJED R,FAILLE C,KALLASSY M,et al.Bacillus cereus biofilms-same, only different[J].Frontiers in Microbiology,2016,7:1 054.
[27] 李燕,李冬冬,陶传敏,等.表皮葡萄球菌生物膜形成及相关基因的检测及评价[J].中华医院感染学杂志, 2010,20(4):473-476.
[28] GRANUM P E,LUND T.Bacillus cereus and its food poisoning toxins[J].FEMS Microbiology Letters,1997, 157(2):223-228.
[29] LOWY F D,HAMMER S M.Staphylococcus epidermidis infections[J].Annals of Internal Medicine,1983,99(6): 834-839.