The subtillicin L produced by Bacillus subtillis DH8043 was systematically purified. Meanwhile, the predicted structural gene and antibacterial properties of Subtillicin L were analyzed in the study. The results showed that one single band with molecular weight at approximately 12.3 kDa was found via tricine sodium dodecyl sulfate poly-acrylamide gel electrophoresis (Tricine-SDS-PAGE) after the purification by anion exchange column and Sephadex-G50 chromatography column from the crude extract precipitated by ammonium sulfate. To identify the similarity of Subtillicin L with other bacteriocins, the analysis and blast of the complete genome sequence were used. The predicted similarity between the amino acid sequence of Subtillicin L and the known amylocyclisin was 99.11%. The main difference between them was at the 88th amino acid by alignment analysis. Finally, antibacterial activity between Subtillicin L and Nisin against Bacillus cereus LMGT2805 was studied. The results showed Subtillicin L can completely inhibit B. cereus LMGT2805 at high concentration, 4 times of minimum inhibition concentration (MIC50), while nisin only prevent the growth of B. cereus LMGT2805 at the same concentration. The antimicrobial results provided more options to inhibit B. cereus besides Nisin.
LI Jinjin
,
WANG Wei
,
TAO Leren
,
CHI Hai
. Purification, identification and antimicrobial characteristics of Subtillicin L produced by Bacillus subtillis DH8043[J]. Food and Fermentation Industries, 2020
, 46(18)
: 8
-12
.
DOI: 10.13995/j.cnki.11-1802/ts.023716
[1] KHOCHAMITN, SIRIPORNADULSILS,SUKON P, et al. Purification, characterization and mode of action of enterocin, a novel bacteriocin produced by Enterococcus faecium TJUQ1[J]. International Journal of Biological Macromolecules, 2020, 144:151-159.
[2] NALISA K, SURASAK S, PEERAPOL S, et al. Antibacterial activity and genotypic-phenotypic characteristics of bacteriocin-producing Bacillus subtilis KKU213:Potential as a probiotic strain[J]. Microbiological Research, 2015, 170:36-50.
[3] 侯养全. 超高压食品灭菌技术研究进展与应用[J]. 山西科技, 2018, 33(1):106-108.
[4] LEE S G, CHANG H C. Purification and characterization of mejucin, a new bacteriocin produced by Bacillus subtilis SN7[J]. LWT-Food Science and Technology, 2018, 87:8-15.
[5] 刘静, 杨富民, 白斌芳, 等. 产细菌素乳酸菌菌株的来源及所产细菌素的分类研究进展[J]. 中国酿造, 2018, 37(6):20-24.
[6] KOMORA N, BRUSCHI C, MAGALHSES R, et al. Survival of Listeria monocytogenes with different antibiotic resistance patterns to food-associated stresses[J]. International Journal of Food Microbiology, 2017, 245(20): 79-87.
[7] 王伟栋, 王丽娟, 刘晓琳, 等. 一起蜡样芽孢杆菌引起食物中毒的实验室检测[J]. 食品界, 2019(2):90-91.
[8] 赵圣明, 赵岩岩, 马汉军.植物乳杆菌JLA-9产细菌素的分离纯化[J].食品与发酵工业,2017,43(6):60-65.
[9] CHI H, HOLO H. Synergistic antimicrobial activity between the broad spectrum bacteriocin garvicin KS and nisin, farnesol and polymyxin b against gram-positive and gram-negative bacteria[J]. Current Microbiology, 2018, 75:272-277.
[10] 张宁, 于功明, 蒋万枫. 曹州木瓜超氧化物歧化酶(SOD)粗酶的提取工艺[J]. 食品安全导刊, 2018(6):105.
[11] PERVMAL V, VENKASTESAN A. Antimicrobial cytotoxic effect and purification of bacteriocin from vancomycin susceptible Enterococcus faecalis and its safety evaluation for probiotization[J]. LWT-Food Science and Technology, 2017,78:303-310.
[12] MAGI A, SEMERARO R, MINGRINO A, et al. Nanopore sequencing data analysis: State of the art, applications and challenges[J]. Brief Bioinform, 2018, 19(6):1 256-1 272.
[13] JAIN M, OLSEN H E, PATEN B, et al. The Oxford Nanopore MinION: Delivery of nanopore sequencing to the genomics community[J]. Genome Biology, 2016, 17(1):239-249.
[14] TARAILO G M, CHEN N. Using RepeatMasker to identify repetitive elements in genomic sequences[J]. Current Protocols in Bioinformatics, 2009, 25:1-14.
[15] HYATT D, CHEN G L, LOCASCIO P F, et al. Prodigal: Prokaryotic gene recognition and translation initiation site identification[J]. BMC Bioinformatics, 2010, 11(1): 119.
[16] HOS SAIN M S,AKHTER M Z,HOSSAIN M M, et al. Complete genome sequence of Bacillus subtilis strain MH1, which has a high level of bacteriocin-like activity, isolated from soil in Bangladesh[J]. Genomics Announcements, 2018, 6(25):1-2.
[17] PEI J J, JIN W G, ELATY A M A, et al. Isolation, purification, and structural identification of a new bacteriocin made by Lactobacillus plantarum found in conventional kombucha[J]. Food Control, 2019,110:106 923.
[18] SCHOLZ R, VATER J, BUDIHARJO A, et al. Amylocyclicin, a novel circular bacteriocin produced by Bacillus amyloliquefaciens FZB42[J]. Journal of Bacteriolog, 2014, 196(10):1 842-1 852.
[19] MOLINATTO G, PUOPOLO G, SONEGO P, et.al. Complete genome sequence of Bacillus amyloliquefaciens subsp plantarum S499, a rhizobacterium that triggers plant defences and inhibits fungal phytopathogens[J]. Journal of Biotechnology, 2016, 238:56-59.
[20] 唐广心,黄薇. 一起蜡样芽孢杆菌引起的持续交叉污染食物中毒流行病学调查[J]. 大家健康(中国版), 2013, 7(7):67-68.
