响应面法优化包被发酵嗜酸乳杆菌的高密度培养
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摘要
针对益生菌嗜酸乳杆菌(Lactobacillus acidophilus)包被发酵的形成特性和营养需要,采用Plackett-Burman试验设计优化其增殖培养基,获得高密度培养条件和提高菌粉存活率方法。结果表明,嗜酸乳杆菌包被发酵的增菌最佳培养基组成为:葡萄糖7.77%、蛋白胨2%、牛肉膏2%、酵母粉3%、柠檬酸氢二铵0.3%、K_2HPO_4·7H_2O 0.2%、MgSO_4·7H_2O 0.09%、MnSO_4·4H_2O 0.025%、NaAc·3H_2O0.3%、吐温-80 0.1%、海藻酸钠1.45%、纳米碳酸钙2.93%。在此优化条件下,嗜酸乳杆菌活菌数达4.32×10~(10)CFU/m L,制备成的冻干粉在37℃条件下贮藏100 d后仍保留75%以上的存活率。
According to the formation characteristics and nutritional requirements of encapsulated fermentation of Lactobacillus acidophilus, the multiplication medium was optimized by Plackett-Burman experimental design and high-density culture condition was obtained and the survival rate of bacteria powder improved. The results showed that the ingredients of the medium for encapsulated fermentation of L. acidophilus were glucose 7.77%,peptone 2%, beef extract 2%, yeast extract powder 3%, diammonium hydrogen citrate 0.3%, K_2HPO_4·7H_2O 0.2%, MgSO_4·7H_2O 0.09%, MnSO_4·4H_2O 0.025%, NaAc·3H_2O 0.3%, Tween-80 0.1%, sodium alginate 1.45%, nano calcium carbonate 2.93%. Under the optimized conditions, the bacterial count of L. acidophilus was 4.32×10~(10) CFU/ml, the survival rate of freeze-dried powder of pre-fermented encapsulated L. acidophilus was more than 75% after storage 100 d at 37 ℃.
According to the formation characteristics and nutritional requirements of encapsulated fermentation of Lactobacillus acidophilus, the multiplication medium was optimized by Plackett-Burman experimental design and high-density culture condition was obtained and the survival rate of bacteria powder improved. The results showed that the ingredients of the medium for encapsulated fermentation of L. acidophilus were glucose 7.77%,peptone 2%, beef extract 2%, yeast extract powder 3%, diammonium hydrogen citrate 0.3%, K_2HPO_4·7H_2O 0.2%, MgSO_4·7H_2O 0.09%, MnSO_4·4H_2O 0.025%, NaAc·3H_2O 0.3%, Tween-80 0.1%, sodium alginate 1.45%, nano calcium carbonate 2.93%. Under the optimized conditions, the bacterial count of L. acidophilus was 4.32×10~(10) CFU/ml, the survival rate of freeze-dried powder of pre-fermented encapsulated L. acidophilus was more than 75% after storage 100 d at 37 ℃.
引文
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[20]张琳,贠婷婷,綦文涛,等.发酵前及发酵后包被粪肠球菌微胶囊抗胁迫作用的研究[J].粮油食品科技,2016,24(2):80-84.
[21]DONNARUMMA G,MOLINARO A,CIMINI D,et al.Lactobacillus crispatus L1:high cell density cultivation and exopolysaccharide structure characterization to highlight potentially beneficial effects against vaginal pathogens[J].BMC Microbiol,2014,14(1):137.
[22]吴军林,柏建玲,莫树平,等.乳酸菌R8高密度培养的发酵工艺研究[J].现代食品科技,2018,34(2):164-170,251.
[23]CHEN H,NIU J,QIN T,et al.Optimization of the medium for Lactobacillus acidophilus by Plackett-Burman and steepest ascent experiment[J].Acta Scient Polon Technol Aliment,2015,14(3):227-232.
[24]?ABUK B,HARSA S,T.Improved viability of Lactobacillus acidophilus NRRL-B 4495 during freeze-drying in whey protein-pullulan microcapsules[J].J Microencapsul,2015,32(3):300-307.
[25]李卫娟,王计伟,王宇霄,等.粪肠球菌耐热型保护剂的筛选及加速储存稳定性研究[J].食品与发酵工业,2015,41(12):144-147.
[2]刘齐,孙美玲,王鲁峰,等.益生菌的功能活性及其分子机理探究的研究进展[J].中国酿造,2014,33(1):14-19.
[3]张宇,王连艳,王艳萍.嗜酸乳杆菌微胶囊制备及工艺优化[J].食品与发酵工业,2018,44(6):132-138.
[4]HOSEINIFAR S H,ROOSTA Z,HAJIMORADLOO A,et al.The effects of Lactobacillus acidophilus as feed supplement on skin mucosal immune parameters,intestinal microbiota,stress resistance and growth performance of black swordtail(Xiphophorus helleri)[J].Fish Shellfish Immunol,2015,42(2):533-538.
[5]LV C H,WANG T,REGMI N,et al.Effects of dietary supplementation of selenium-enriched probiotics on production performance and intestinal microbiota of weanling piglets raised under high ambient temperature[J].J Anim Physiol Anim Nutr,2015,99(6):1161-1171.
[6]MENG J,ZHANG Q X,LU R R.Surface layer protein from Lactobacillus acidophilus,NCFM inhibit intestinal pathogen-induced apoptosis in HT-29cells[J].Int J Biol Macromol,2017,96(3):766-774.
[7]PACHECO ORDAZ R,WALL MEDRANO A,GO譙I M G,et al.Effect of phenolic compounds on the growth of selected probiotic and pathogenic bacteria[J].Lett Appl Microbiol,2018,66(1):25-31.
[8]WEN K,LI G,BUI T,et al.High dose and low dose Lactobacilli acidophilus exerted differential immune modulating effects on T cell immune responses induced by an oral human rotavirus vaccine in gnotobiotic pigs[J].Vaccine,2012,30(6):1198-1207.
[9]HASANI S,SARI A A,HESHMATI A,et al.Physicochemical and sensory attributes assessment of functional low-fat yogurt produced by incorporation of barley bran and Lactobacillus acidophilus[J].Food Sci Nutr,2017,5(4):875-880.
[10]ZARRATI M,SALEHI E,NOURIJELYANI K,et al.Effects of probiotic yogurt on fat distribution and gene expression of proinflammatory factors in peripheral blood mononuclear cells in overweight and obese people with or without weight-loss diet[J].J Am Coll Nutr,2014,33(6):417-425.
[11]ZHANG L,LI J,YUN T T,et al Effects of pre-encapsulated and pro-encapsulated Enterococcus faecalis on growth performance,blood characteristics,and cecal microflora in broiler chickens[J].Poultry Sci,2015,94(11):2821-2830.
[12]潘秋月,高红岩,南天月,等.益生菌的微胶囊研究进展[J].食品科技,2014,39(7):31-36.
[13]GORANOV B,BLAZHEVA D,KOSTOV G,et al.Lactic acid fermentation with encapsulated Lactobacillus casei ssp.rhamnosus ATCC 11979(NBIMCC 1013)in alginate/chitosan matrices[J].Bulgar J Agr Sci,2013,19(2):101-104.
[14]秦腾飞,贠婷婷,王春玲,等.发酵前包被布拉迪酵母生长代谢及其抗逆性机理[J].食品科学,2017,38(22):137-14.
[15]OZYURT V H,魨TLES S.Properties of probiotics and encapsulated probiotics in food[J].Acta Scient Polon Technol Aliment,2014,13(4):413-424.
[16]张娅,冷一非,李建华,等.响应面法优化嗜酸乳杆菌增殖培养基[J].中国酿造,2011,30(9):134-137.
[17]陈婷,王玉华,蔡丹,等.益生菌微胶囊技术研究进展[J].中国乳品工业,2016,44(1):31-37.
[18]宋娇娇,包秋华,王亚利,等.瑞士乳杆菌MG9-2微胶囊的制备工艺优化及其性能分析[J].食品科学,2017,38(14):49-57.
[19]贠婷婷.饲用益生菌保护技术的研究进展[C].2017第二届中国畜牧生物科技大会论文集.北京:中国农业科学院饲料研究所、第十五届中国畜牧业博览会组委会、新饲料传媒,2017.
[20]张琳,贠婷婷,綦文涛,等.发酵前及发酵后包被粪肠球菌微胶囊抗胁迫作用的研究[J].粮油食品科技,2016,24(2):80-84.
[21]DONNARUMMA G,MOLINARO A,CIMINI D,et al.Lactobacillus crispatus L1:high cell density cultivation and exopolysaccharide structure characterization to highlight potentially beneficial effects against vaginal pathogens[J].BMC Microbiol,2014,14(1):137.
[22]吴军林,柏建玲,莫树平,等.乳酸菌R8高密度培养的发酵工艺研究[J].现代食品科技,2018,34(2):164-170,251.
[23]CHEN H,NIU J,QIN T,et al.Optimization of the medium for Lactobacillus acidophilus by Plackett-Burman and steepest ascent experiment[J].Acta Scient Polon Technol Aliment,2015,14(3):227-232.
[24]?ABUK B,HARSA S,T.Improved viability of Lactobacillus acidophilus NRRL-B 4495 during freeze-drying in whey protein-pullulan microcapsules[J].J Microencapsul,2015,32(3):300-307.
[25]李卫娟,王计伟,王宇霄,等.粪肠球菌耐热型保护剂的筛选及加速储存稳定性研究[J].食品与发酵工业,2015,41(12):144-147.