植物乳杆菌LPL-1产细菌素发酵培养基优化
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摘要
乳酸细菌素是细胞在转录翻译过程中通过核糖体机制合成,并分泌到菌体体外的一类具有抑菌活性的蛋白质,细菌素具有无抗药性、无毒副作用及易被人体降解的特点,因此是一种天然的食品防腐保鲜剂。为了提高植物乳杆菌LPL-1所产细菌素的产量,以单增李斯特菌为指示菌,相对抑菌效价为效应值,通过响应面法对发酵培养基组成进行优化,确定了最优培养基组成。利用单因素试验与Plackett-Burman试验,确定了主要影响因素为葡萄糖质量分数、胰蛋白胨质量分数与吐温-80体积比,最陡爬坡试验与Box-Behnken响应面试验确定了最优培养基组成为:葡萄糖质量分数2.08%、酵母粉质量分数0.51%、胰蛋白胨质量分数1.02%、牛肉膏质量分数1%、吐温-80体积比1.02 mL/L、磷酸氢二钾质量浓度3 g/L、乙酸钠质量分数0.5%、硫酸镁质量浓度0.2 g/L、硫酸锰质量浓度0.3g/L、柠檬酸氢二铵质量浓度2 g/L、蒸馏水体积1 L。在此条件下细菌素效价(752.11 AU/mL)比优化前(286.67AU/mL)提高了1.62倍。因此,发酵培养基的优化为菌种与细菌素的产业化应用奠定了基础。
Bacteriocins from lactic acid bacteria(LAB) are ribosomally synthesized peptides or proteins that inhibit or kill the closely related species. Based on the merits of easy degradation by proteases,nonresistance and non-toxic side effects, plantaricin LPL-1 is a promising natural and safe biological preservative for food preservation industry. Aiming at the improvement of plantaricin LPL-1 fromLactobacillus plantarumLPL-1,the inhibitory activity(AU/mL) against Listeria monocytogenes 54002 was used as effect index,and the optimized medium composition was determined by response surface methodology. The major factors were determined as glucose,yeast extract and Tween-80 by single factor tests and Plackett-Burman tests. Based on the results of the steepest grade test and response surface methodology,the optimal medium composition was followed as glucose 2. 08%,yeast extract 0. 51%,tryptone 1. 02%,beef extract 1%,Tween-80 1. 02 mL/L,K_2HPO_4 3 g/L,NaAc 0. 5%,MgSO_4 0. 2 g/L,MnSO_4 0. 3 g/L,tri-ammonium citrate 2 g/L,and dd H_2O 1 L. Under the optimal conditions,the inhibitory activity reached up to 752. 11 AU/mL which was increased by 1. 62 times. Finally,the research provided insight into the potential use of bacteriocin as a food preservative in the food industry.
Bacteriocins from lactic acid bacteria(LAB) are ribosomally synthesized peptides or proteins that inhibit or kill the closely related species. Based on the merits of easy degradation by proteases,nonresistance and non-toxic side effects, plantaricin LPL-1 is a promising natural and safe biological preservative for food preservation industry. Aiming at the improvement of plantaricin LPL-1 fromLactobacillus plantarumLPL-1,the inhibitory activity(AU/mL) against Listeria monocytogenes 54002 was used as effect index,and the optimized medium composition was determined by response surface methodology. The major factors were determined as glucose,yeast extract and Tween-80 by single factor tests and Plackett-Burman tests. Based on the results of the steepest grade test and response surface methodology,the optimal medium composition was followed as glucose 2. 08%,yeast extract 0. 51%,tryptone 1. 02%,beef extract 1%,Tween-80 1. 02 mL/L,K_2HPO_4 3 g/L,NaAc 0. 5%,MgSO_4 0. 2 g/L,MnSO_4 0. 3 g/L,tri-ammonium citrate 2 g/L,and dd H_2O 1 L. Under the optimal conditions,the inhibitory activity reached up to 752. 11 AU/mL which was increased by 1. 62 times. Finally,the research provided insight into the potential use of bacteriocin as a food preservative in the food industry.
引文
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12吕燕妮,李平兰,周伟.戊糖乳杆菌31-1菌株产细菌素发酵条件优化[J].微生物学通报,2005,32(3):13-19.LYanni,LI Pinglan,ZHOU Wei.Optimization of bacteriocin produced by Lactobacillus pentosus 31-1[J].Microbiology China,2005,32(3):13-19.(in Chinese)
13 TODOROV S D,DICKS L M.Effect of medium components on bacteriocin production by Lactobacillus plantarum strains ST23LDand ST341LD,isolated from spoiled olive brine[J].Microbiological Research,2006,161(2):102.
14 HALAMI P M,CHANDRASHEKAR A.Enhanced production of pediocin C20 by a native strain of Pediococcus acidilactici C20 in an optimized food-grade medium[J].Process Biochemistry,2005,40(5):1835-1840.
15 RATNAM B V V,NARASIMHA R M,DAMODAR R M,et al.Optimization of fermentation conditions for the production of ethanol from sago starch using response surface methodology[J].World Journal of Microbiology and Biotechnology,2003,19(5):523-526.
16朱凤,陈长华,张琪.吐温-85对螺旋霉素发酵过程的影响[J].中国医药工业杂志,2008,39(3):176-178.
17 GARVER K I,MURIANA P M.Detection,identification and characterization of bacteriocin-producing lactic acid bacteria from retail food products[J].International Journal of Food Microbiology,1993,19(4):241-258.
18庄绪亮,张洪勋,马桂荣,等.不同发酵条件下Tween80对乳酸链球菌SM526产Nisin的影响[J].过程工程学报,2000,21(2):145-148.
19沈慧,秦海宏,王福俤,等.低锌浓度培养基对Caco2细胞二价金属离子转运体mRNA表达的影响[J].中华预防医学杂志,2006,40(1):55-57.
20谢英,覃倩倩,张京声,等.植物乳杆菌LB-B1产细菌素发酵条件的优化[J].中国酿造,2010,29(10):22-25.
2 YANG S C,LIN C H,SUNG C T,et al.Corrigendum:antibacterial activities of bacteriocins:application in foods and pharmaceuticals[J].Frontier in Microbiology,2014,5(22):683.
3 COTTER P D.Bacteriocins-a viable alternative to antibiotics?[J].Nature Reviews Microbiology,2013,11(2):95-105.
4 WANG Y,SHANG N,QIN Y,et al.The complete genome sequence of Lactobacillus plantarum LPL-1,a novel antibacterial probiotic producing class IIa bacteriocin[J].Journal of Biotechnology,2018,266:84-88.
5 LIU G,REN L,SONG Z,et al.Purification and characteristics of bifidocin A,a novel bacteriocin produced by Bifidobacterium animals BB04 from centenarians'intestine[J].Food Control,2015,50:889-895.
6 GUI M,ZHAO B,SONG J,et al.Paraplantaricin L-ZB1,a novel bacteriocin and its application as a biopreservative agent on quality and shelf life of rainbow trout fillets stored at 4℃[J].Applied Biochemistry&Biotechnology,2014,174(6):2295-2306.
7 DALIE D K D,DESCHAMPS A M,RICHARDFORGET F.Lactic acid bacteria-potential for control of mould growth and mycotoxins:a review[J].Food Control,2010,21(4):370-380.
8 KANMANI P,SATISH K R,YUVARAJ N,et al.Probiotics and its functionally valuable products-a review[J].Critical Reviews in Food Science&Nutrition,2013,53(6):641-658.
9 DEEGAN L H,COTTER P D,HILL C,et al.Bacteriocins:biological tools for bio-preservation and shelf-life extension[J].International Dairy Journal,2006,16(9):1058-1071.
10 ABBASILIASI S,TAN J S,IBRAHIM T A T,et al.Fermentation factors influencing the production of bacteriocins by lactic acid bacteria:a review[J].RSC Advances,2017,7:29395-29420.
11 DELAADO A,BRITO D,FEVEREIRO P,et al.Bioactivity quantification of crude bacteriocin solutions[J].Journal of Microbiological Methods,2005,62(1):121-124.
12吕燕妮,李平兰,周伟.戊糖乳杆菌31-1菌株产细菌素发酵条件优化[J].微生物学通报,2005,32(3):13-19.LYanni,LI Pinglan,ZHOU Wei.Optimization of bacteriocin produced by Lactobacillus pentosus 31-1[J].Microbiology China,2005,32(3):13-19.(in Chinese)
13 TODOROV S D,DICKS L M.Effect of medium components on bacteriocin production by Lactobacillus plantarum strains ST23LDand ST341LD,isolated from spoiled olive brine[J].Microbiological Research,2006,161(2):102.
14 HALAMI P M,CHANDRASHEKAR A.Enhanced production of pediocin C20 by a native strain of Pediococcus acidilactici C20 in an optimized food-grade medium[J].Process Biochemistry,2005,40(5):1835-1840.
15 RATNAM B V V,NARASIMHA R M,DAMODAR R M,et al.Optimization of fermentation conditions for the production of ethanol from sago starch using response surface methodology[J].World Journal of Microbiology and Biotechnology,2003,19(5):523-526.
16朱凤,陈长华,张琪.吐温-85对螺旋霉素发酵过程的影响[J].中国医药工业杂志,2008,39(3):176-178.
17 GARVER K I,MURIANA P M.Detection,identification and characterization of bacteriocin-producing lactic acid bacteria from retail food products[J].International Journal of Food Microbiology,1993,19(4):241-258.
18庄绪亮,张洪勋,马桂荣,等.不同发酵条件下Tween80对乳酸链球菌SM526产Nisin的影响[J].过程工程学报,2000,21(2):145-148.
19沈慧,秦海宏,王福俤,等.低锌浓度培养基对Caco2细胞二价金属离子转运体mRNA表达的影响[J].中华预防医学杂志,2006,40(1):55-57.
20谢英,覃倩倩,张京声,等.植物乳杆菌LB-B1产细菌素发酵条件的优化[J].中国酿造,2010,29(10):22-25.