Screening of a Weakly Post-acidified Lactobacillus bulgaricus KLDS1.1011 and Its Genome-wide Annotation
-
摘要: 为了研究影响保加利亚乳杆菌后酸化的关键基因，为酸奶发酵剂的开发提供分子水平的理论基础。本实验以实验室现有的8株保加利亚乳杆菌作为出发菌株，通过对其生长性能以及对酸敏感性筛选出KLDS1.0207、KLDS1.0205、KLDS1.1001、KLDS1.1011产酸差异性明显的4株保加利亚乳杆菌，通过对4株保加利亚乳杆菌单菌株发酵乳的凝乳时间、滴定酸度、乳糖消耗进行检测，分析得到菌株KLDS1.1011后酸化能力最弱。通过Illumina HiSeq与Illumina MiSeq测序平台对菌株菌株KLDS1.1011进行基因组测序，得到KLDS1.1011基因组全长1887491 bp，平均G+C含量为39.83%，基因组中共预测出2098个CDS，其总长度为1622760 bp，编码区域总长度占全基因组比例85.97%，编码基因的平均长度为773 bp；利用同源聚类分析方式比较保加利亚乳杆菌KLDS1.1011和KLDS1.0207基因组，发现两者有1631个共有基因，KLDS1.1011有353个特有基因，KLDS1.0207有320个特有基因，进而通过对特有基因进行KEGG通路的注释以及后酸化相关通路的分析得到6个与后酸化相关的基因，1.1011_GM000805、1.1011_GM002068、1.1011_GM000803、1.1011_GM000804四个基因是关于生物膜的形成，菌株的物质转运、1.1011_GM000260基因属于丙酮酸代谢通路，是乳酸生成过程的重要通路、1.1011_GM000194基因是蛋白水解的关键酶基因。在基因水平上为菌株KLDS1.1011较弱的后酸化特性提供了重要的理论依据。Abstract: In order to study the key genes that affect the post-acidification of Lactobacillus bulgaricus, it provides a theoretical basis for the development of yogurt starter at the molecular level. In this experiment, 8 existing Lactobacillus bulgaricus in the laboratory were used as starting strains. Based on their growth performance and acid sensitivity, 4 strains of Lactobacillus bulgaricus(KLDS1.0207, KLDS1.0205, KLDS1.1001, and KLDS1.1011) with obvious differences in acid production were selected. Four strains of Lactobacillus bulgaricus were tested for the curd time, titrated acidity, and lactose consumption of four strains of Lactobacillus bulgaricus single strains. The analysis showed that the strain KLDS1.1011 had the weakest acidification capacity.Genome sequencing of strain KLDS1.1011 via Illumina HiSeq and Illumina MiSeq sequencing platform. The total length of the KLDS1.1011 genome was 1884491 bp.The average G+C content was 39.83%. A total of 2098 CDS were predicted in the genome, with a total length of 1622760 bp.The total length of the coding region accounted for 85.97% of the whole genome and the average length of the coding gene was 773 bp. The genomes of Lactobacillus bulgaricus KLDS1.1011 and KLDS1.0207 were used homology cluster analysis to compare. It was found that there were 1631 genes in common, KLDS1.1011 had 353 unique genes, and KLDS1.0207 had 320 unique genes. Furthermore, by annotating the KEGG pathway and analyzing the post-acidification related pathways of the unique genes, 6 genes related to post-acidification were obtained. The four genes 1011_GM000805, 1.1011_GM002068, 1.1011_GM000803, and 1.1011_GM000804 were related to the formation of biofilms and the material transport of strains. 1.1011_GM000260 gene belonged to the pyruvate metabolism pathway and was an important pathway in the lactic acid production process and the 1.1011_GM000194 gene was a key enzyme gene for proteolysis. It would provide an important theoretical basis for the weak post-acidification characteristics of strain KLDS1.1011 at the gene level.
-
Keywords:
- Lactobacillus bulgaricus /
- screening /
- post-acidification /
- genome-wide annotation /
- unique genes
-
[1] 郭清泉,张兰威,夏秀芳. 酸奶制品发生后酸化主要发酵剂菌确定及性质研究[J]. 食品与发酵工业,2002,28(4):24-27. [2] 徐成勇,吴昊,郑思聪,等.酸乳后酸化影响因子的初步研究[J]. 食品与发酵工业,2006,32(12):10-14. [3] C.Beal,J.Skokanova,et al. Combined effects of culture conditions and storage time on acidification and viscosity of stirred yogurt[J]. Journal of Dairy Science,1999,82(4):673-681.
[4] Ongol M P,Sawatari Y,Ebina Y,et al. Yoghurt fermented by Lactobacillus delbrueckii subsp. bulgaricus H+-ATPase-defective mutants exhibits enhanced viability of Bifidobacterium breve during storage[J]. International Journal of Food Microbiology,2007,116(3):358-366.
[5] Chen M J,Tang H Y,Chiang M L. Effects of heat,cold,acid and bile salt adaptations on the stress tolerance and protein expression of kefir-isolated probiotic,Lactobacillus kefiranofaciens M1[J]. Food Microbiology,2017,66:20-27.
[6] Rivals J P,Catherine Béal,Thammavongs B,et al. Cryotolerance of Lactobacillus delbrueckii subsp. bulgaricus CFL1 is modified by acquisition of antibiotic resistance[J]. Cryobiology,2007,55(1):19-26.
[7] Li C,Song J,Kwok L Y,et al. Influence of,Lactobacillus plantarum,on yogurt fermentation properties and subsequent changes during postfermentation storage[J]. Journal of Dairy Science,2017,100(4):2512-2525.
[8] Settachaimongkon S,Van Valenberg H J F,Gazi I,et al. Influence of Lactobacillus plantarum WCFS1 on post-acidification,metabolite formation and survival of starter bacteria in set-yoghurt[J]. Food Microbiology,2016,59:14-22.
[9] Wang T,Xu Z,Lu S,et al. Effects of glutathione on acid stress resistance and symbiosis between Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus[J]. International Dairy Journal,2016,61:22-28.
[10] Elisabetta Benozzi,Romano A,Capozzi V,et al. Monitoring of lactic fermentation driven by different starter cultures via direct injection mass spectrometric analysis of flavour-related volatile compounds[J]. Food Research International,2015,76:682-688.
[11] Indyk H E,Edwards M J,Woollard D C. High performance liquid chromatographic analysis of lactose-hydrolysedmilk[J]. Food Chemistry,1996,57(4):575-580.
[12] 丁秀云,蒙月月,李柏良,等. 嗜热链球菌KLDSSM胞外多糖生物合成途径的分析[J].食品工业科技,2017(12):157-162. [13] 张文羿,孟和,张和平. 乳酸菌基因组学研究进展[J].微生物学报,2008,48(9):1270-1275. [14] Van d G M,Penaud S,Grimaldi C,et al. The complete genome sequence of Lactobacillus bulgaricus reveals extensive and ongoing reductive evolution[J]. Proceedings of the National Academy of Sciences of the United States of America,2006,103(24):9274-9279.
[15] Tanigawa K,Watanabe K. Multilocus sequence typing reveals a novel subspeciation of Lactobacillus delbrueckii[J]. Microbiology,2011,157(3):727-738.
[16] Song Y,Sun Z,Guo C,et al. isolated from naturally fermented dairy foods[J]. Scientific Reports,2016,6:22704.
[17] 程娜. 基于组学技术解析保加利亚乳杆菌LJJ的耐酸机制[D].临汾:山西师范大学,2018. [18] 孙懿琳,田辉,方伟,等.保加利亚乳杆菌关键糖代谢机制的研究[J]. 食品工业科技,2012,33(22):202-205. [19] 李柏良,丁秀云,靳妲,等. 基于基因组学分析嗜热链球菌KLDS SM的蛋白质水解系统和氨基酸合成途径[J].食品科学,2018(1):120-126. [20] Feng Q,Huang T,Zhao Q,et al. Analysis of collinear regions of Oryza AA and CC genomes[J]. Journal of Genetics and Genomics,2009,36(11):667-677.
[21] Li B,Ding X,Evivie S E,et al. Short communication:Genomic and phenotypic analyses of exopolysaccharides produced by,Streptococcus thermophilus,KLDS SM[J]. Journal of Dairy Science,2017,101(1):106-112.
[22] Hui T,Bailiang L,Smith E,et al. Technological and genomic analysis of roles of the cell-envelope protease PrtS in yoghurt starter development[J]. International Journal of Molecular Sciences,2018,19(4):1068.
[23] Salmon-Divon M,Dvinge H,Tammoja K,et al. PeakAnalyzer:Genome-wide annotation of chromatin binding and modification loci[J]. BMC Bioinformatics,2010,11(1):415.
[24] Schmid R,Blaxter M L. Annot8r:GO,EC and KEGG annotation of EST datasets[J]. Bmc Bioinformatics,2008,9(1):180.
[25] Galperin M Y,Makarova K S,Wolf Y I,et al. Expanded microbial genome coverage and improved protein family annotation in the COG database[J]. Nucleic Acids Research,2015,43(D1):D261-D269.
[26] Hickey M W,Hillier A J,Jago G R. Transport and metabolism of lactose,glucose,and galactose in homofermentative lactobacilli[J]. Applied & Environmental Microbiology,1986,51(4):825-831.
[27] Aat Ledeboer,Jeroen Hugenholtz,Jan Kok,et al. Thirty years of research on lactic acid bacteria[M]. Holland:Max Blngham,2011:493-535.
[28] 韩巍巍,侯俊财,曹秋阁,等. 不同生长阶段保加利亚乳杆菌关键蛋白酶基因表达变化规律[J]. 食品与发酵工业,2014,40(3):13-19. [29] Leong-Morgenthaler,P,Zwahlen,M C,Hottinger,H. Lactose metabolism in Lactobacillus bulgaricus:Analysis of the primary structure and expression of the genes involved[J]. Journal of Bacteriology,1991,173(6):1951-1957.
[30] Fernandez A,Ogawa J,Stéphanie Penaud,et al. Rerouting of pyruvate metabolism during acid adaptation in Lactobacillus bulgaricus[J]. Proteomics,2008,8(15):3154-3163.
-
期刊类型引用(4)
1. 舒明，潘凡达，边腾飞，袁凯龙，项波卡，张勇刚，杨洋，钟卫鸿. 造纸法再造烟叶浓缩液中抗逆酵母的分离鉴定与代谢活性研究. 轻工学报. 2023(01): 54-62 . 
百度学术
2. 张蓝月，孙万成，罗毅皓. 基于非靶向代谢组学分析不同地区羊肉代谢物的差异. 现代食品科技. 2023(07): 91-101 . 百度学术
3. 赵电波，马燕青，王少丹，王雯雯. 基于非靶向代谢组学解析肉桂精油纳米乳抑制假单胞菌CM2作用机制研究. 食品工业科技. 2023(17): 168-175 . 本站查看
4. 玉霞，赵飞燕，孙志宏. 肠道中有益菌培养组学的研究进展. 食品科学. 2023(23): 365-371 . 百度学术
其他类型引用(5)
计量
- 文章访问数: 271
- HTML全文浏览量: 45
- PDF下载量: 31
- 被引次数: 9
下载:
