枯草芽孢杆菌YT168-6产抗菌肽sublancin的发酵工艺优化及培养基筛选的研究

doi:10.16431/j.cnki.1671-7236.2021.09.014

Abstract

Abstract: The aim of this study was to research the optimal fermentation conditions and culture medium component for sublancin, an antimicrobial peptide produced by Bacillus subtilis YT168-6, in order to increase the content of sublancin in fermentation broth. A single factor test was used to screen out the optimal fermentation conditions, inorganic salts, carbon and nitrogen sources of fermentation medium. The optimum fermentation medium component was determined by the orthogonal test, the steepest climbing test and the response surface analysis method of Box-Behnken design. The results showed that the optimal fermentation conditions for Bacillus subtilis YT168-6 to produce the antimicrobial peptide sublancin were 1.0% bacterial inoculum, fermentation temperature 37 ℃ and initial pH 7.0, the optimum fermentation medium was K₂HPO₄ 5 g/L, KH₂PO₄ 5 g/L, MgSO₄ 10 g/L, soluble starch 30.2 g/L, peptone 23.6 g/L and corn steep liquor 18.2 g/L. After optimization, the yield of sublancin was increased from 756 μg/mL to 1 581 μg/mL, which was 2.09 times as high as before optimization. In summary, the optimal fermentation conditions and culture medium selected in this research laid the technical foundation for the subsequent industrial production of Bacillus subtilis YT168-6.

Key words: Bacillus subtilis YT168-6; fermentation technology; sublancin

CLC Number:

S816.7

LIU Yangke, ZHAO Tianxiao, LU Xiaoying, LI Lei, CHENG Peng, LI Songjian, XIE Shunchang. Optimization of Fermentation Technology and Selection of Culture Medium for Antimicrobial Peptide Sublancin Produced by Bacillus subtilis YT168-6[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3232-3241.

References

[1] 翟培, 韩晋辉, 潘晓瑜.基于天蚕素A和胡蜂毒素的杂合肽设计及活性鉴定[J]. 中国畜牧兽医, 2020, 47(9):3006-3013. ZHAI P, HAN J H, PAN X Y.Design and activity identification of hybrid peptide based on cecropin A and mastoparan[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(9):3006-3013.(in Chinese)
[2] ZHANG L J, GALLO R L.Antimicrobial peptides[J]. Current Biology, 2016, 26(1):R14-R19.
[3] 聂守民, 赵瑞君, 杜斌, 等.抗菌肽对肿瘤细胞的作用机制及应用前景[J]. 国际医学寄生虫病杂志, 2015, 42(1):44-48. NIE S M, ZHAO R J, DU B, et al. The mechanism of action and application prospects of antibacterial peptide on tumor cell[J]. International Journal of Medical Parasitic Diseases, 2015, 42(1):44-48.(in Chinese)
[4] 卢军霞, 张宁, 王娟, 等.抗菌肽的作用机制及在畜禽生产中的应用[J]. 中国饲料, 2020, 4:20-22. LU J X, ZHANG N, WANG J, et al. Action mechanism of antibacterial peptide and its application in livestock and poultry production[J]. China Feed, 2020, 4:20-22.(in Chinese)
[5] 王臣, 廖成水, 牛明福, 等.Metalnikowin类抗菌肽MLP的设计及其抗菌活性的测定[J]. 中国兽医科学, 2016, 46(7):841-846. WANG C, LIAO C S, NIU M F, et al. Molecular design and antibacterial activity determination of Metalnikowin-like antibacterial peptide[J]. Chinese Veterinary Science, 2016, 46(7):841-846.(in Chinese)
[6] VAN BOECKEL T P, PIRES J, SILVESTER R, et al. Global trends in antimicrobial resistance in animal in low and middle-income countries[J]. Science, 2019, 365(6459):1944.
[7] BAKKEREN E, HUISMAN J S, FATTINGER S A, et al. Salmonella persisters promote the spread of antibiotic resistance plasmids in the gut[J]. Nature, 2019, 573(7773):276-280.
[8] PIRES J, SIRWARDENA T N, STACH M, et al. In vitro activity of the novel antimicrobial peptide dendrimer G3KL against multidrug-resistant Acinetobacter baumannii and Pseudomonas aeruginosa[J]. Anitimi-crobial Agents and Chemotherapy, 2015, 59(12):7915-7918.
[9] 汪以真.动物源抗菌肽的研究现状和展望[J]. 动物营养学报, 2014, 26(10):2934-2941. WANG Y Z.Antimicrobial peptides of animal origin:Current situation and prospect[J]. Chinese Journal of Animal Nutrition, 2014, 26(10):2934-2941.(in Chinese)
[10] 刘伦伦, 刘焱, 瞿朝霞, 等.抗菌肽的功能及生产研究进展[J]. 中国酿造, 2015, 34(8):1-4. LIU L L, LIU Y, QU Z X, et al. Research progress of function and production of antibacterial peptides[J]. China Brewing, 2015, 34(8):1-4.(in Chinese)
[11] MANSOUR S C, PENA O M, HANCOCK R E.Host defense peptides:Front-line immunomodulators[J]. Trends Immunology, 2014, 35(9):443-450.
[12] 黄佳明, 姜宁, 张爱忠.基因工程菌生产抗菌肽的研究进展[J]. 微生物学通报, 2019, 46(3):654-659. HUANG J M, JIANG N, ZHANG A Z.Progress in engineering bacteria producing antibacterial peptides[J]. Microbiology China, 2019, 46(3):654-659.(in Chinese)
[13] PAIK S H, CHAKICHERLA A, HANSEN J N.Identification and characterization of the structural and transporter genes for, and the chemical and biological properties of, sublancin 168, a novel lantibiotic produced by Bacillus subtilis 168[J]. Journal of Biological Chemistry, 1998, 273(36):23134-23142.
[14] 李尧, 陈婷婷.毕赤酵母基因工程菌发酵生产抗菌肽工艺研究[J]. 食品与发酵科技, 2017, 53(5):17-20. LI Y, CHEN T T.Optimization of the fermentional process of expression of antibacterial peptide in Pichia pastoris[J]. Food and Fermentation Sciences & Technology, 2017, 53(5):17-20.(in Chinese)
[15] 翟立翔, 李国丽, 冷艳, 等.产抗菌肽PSI的重组乳酸克鲁维酵母高密度发酵及产物的分离纯化研究[J]. 食品与发酵工业, 2018, 44(4):13-21. ZHAI L X, LI G L, LENG Y, et al. High density fermentation of recombinant Kluyveromyces lactis producing antimicrobial peptide PSI and purification of its products[J]. Food and Fermentation Industries, 2018, 44(4):13-21.(in Chinese)
[16] 张雯, 卞丹, 沈燕秋, 等.枯草芽孢杆菌抑菌活性物质鉴定、抑菌特性及发酵条件优化[J]. 中国食品学报, 2017, 17(12):105-114. ZHANG W, BIAN D, SHEN Y Q, et al. Identification and characterization of antibacterial metabolites and optimization of cultural conditions for Bacillus subtilis[J]. Journal of Chinese Institute of Food Science and Technology, 2017, 17(12):105-114.(in Chinese)
[17] 张志焱, 赵倩, 于佳民, 等.一株枯草芽孢杆菌产抗菌肽培养基筛选及发酵工艺优化的研究[J]. 中国畜牧兽医, 2019, 46(4):1217-1226. ZHANG Z Y, ZHAO Q, YU J M, et al. Selection of culture medium and optimization of fermention conditions for Bacillus subtilis producing antimicrobial peptides[J]. China Animal Husbandry & Veterinary Medicine, 2019, 46(4):1217-1226.(in Chinese)
[18] SAIER M H, JR CHAUVAUX S, DEUTSCHER J, et al. Protein phosphorylation and regulation of carbon metabolism in Gram-negative versus Gram-positive bacteria[J]. Trends in Biochemical Sciences, 1995, 20(7):267-271.
[19] 李鸿雁, 陆健, 李晓敏.阿拉伯呋喃糖苷酶的重组表达及其发酵工艺优化[J]. 食品与发酵工业, 2020, 46(15):14-20. LI H Y, LU J, LI X M.Recombinant expression and fermentation optimization of arabinofurosidase from Penicillium oxalicum[J]. Food and Fermentation Industries, 2020, 46(15):14-20.(in Chinese)
[20] 吴耀辉, 姜伦, 褚磊, 等.抗菌肽生产工艺研究[M].国外医药抗生素分册, 2017, 38(1):33-37. WU Y H, JIANG L, CHU L, et al. Study on Production Technology of Antibacterial Peptide[M].World Notes on Antibiotics, 2017, 38(1):33-37.(in Chinese)
[21] 应晓彩, 刘磊, 裴志花, 等.动物血缘抗菌肽的研究进展及应用[J]. 动物医学进展, 2020, 41(9):115-118. YING X C, LIU L, PEI Z H, et al. Progress and application of animal blood-derived antibacterial peptides[J]. Progress in Veterinary Medicine, 2020, 41(9):115-118.(in Chinese)
[22] 李波.天蚕素的规模化制备及其在断奶仔猪日粮中的应用研究[D].长春:吉林大学, 2015. LI B.Study of large-scale preparation of cecropin and it's utilization in weaniling pig diets[D].Changchun:Jilin University, 2015.(in Chinese)
[23] 姬生跃.新型抗菌糖肽-sublancin的增量表达及其生物学活性研究[D].杨凌:西北农林科技大学, 2015. JI S Y.Producion improvement of novel antimicrobial glycopeptide sublancin and its bioactivity study[D].Yangling:Northwest A&F University, 2015.(in Chinese)

Optimization of Fermentation Technology and Selection of Culture Medium for Antimicrobial Peptide Sublancin Produced by Bacillus subtilis YT168-6

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