响应面法优化丁酸梭菌快速增殖基础培养基Optimization of Basic Medium for Clostridium butyricum Rapid Proliferation by Response Surface Methodology
刘瑾,赵华
摘要(Abstract):
丁酸梭菌是一种专性厌氧菌,具有极强的整肠作用,在食品、医药、发酵工业等多个领域中有着十分广阔的应用前景。但是由于培养条件苛刻,培养物活菌数量少,其多种应用被限制。通过单因素实验研究了5种碳源、5种氮源、4种除氧剂以及培养温度对丁酸梭菌生长的影响,并在最陡爬坡实验的基础上使用Design Expert 10.0软件研究各因素对丁酸梭菌OD值的响应面分析,最终确定最优培养基。结果表明,优化后培养基中最佳碳源为葡萄糖(14.0 g/L),最佳氮源为酵母膏(16.0 g/L),最佳除氧剂为半胱氨酸盐酸盐(0.9 g/L),最适培养温度为37℃。此时丁酸梭菌生长最快速,且最大OD_(600)值达到3.350 4,比优化前提高约132.72%。
关键词(KeyWords): 响应面分析;丁酸梭菌;菌体浓度;单因素实验
基金项目(Foundation):
作者(Author): 刘瑾,赵华
参考文献(References):
- [1]LIU X.Production of butyric acid and hydrogen by metabolically engineered mutants of Clostridium tyrobutyricum[D].Columbus:the Ohio State University,2005.
- [2]孔青.丁酸梭菌培养与发酵动力学以及调节腹泻小鼠肠道菌群平衡的研究[D].杭州:浙江大学,2006.
- [3]WEIMER P J,MOEN G N.Quantitative analysis of growth and volatile fatty acid production by the anaerobic ruminal bacterium Megasphaera elsdenii T81[J].Applied Microbiology & Biotechnology,2013,97(9):4075-4081.
- [4]蒋维勇.丁酸梭菌BJ-10产芽孢条件优化及放大培养工艺研究[D].长沙:湖南农业大学,2019.
- [5]TRAN N T,LI Z Z,MA H Y,et al.Clostridium butyricum:a promising probiotic confers positive health benefits in aquatic animals[J].Reviews in Aquaculture,2020,12(4):2573-2589.
- [6]张倩.丁酸钠在动物生产中的应用研究进展[J].中国饲料,2009(11):28-30.
- [7]WYDAU-DEMATTEIS S,LOUIS M,ZAHR N,et al.The functional dlt operon of Clostridium butyricum controls the D-alanylation of cell wall components and influences cell septation and vancomycin-induced lysis[J].Anaerobe,2015,35:105-114.
- [8]梁静,聂存喜,张文举,等.丁酸梭菌的生物学功能及其在动物生产中的应用[J].动物营养学报,2018,30(5):1639-1646.
- [9]HAGIHARA M,ARIYOSHI T,KUROKI Y,et al.Clostridium butyricum enhances colonization resistance against Clostridioides difficile by metabolic and immune modulation[J].Scientific Reports,2021,11(1):15007.
- [10]LIU S Q,BISCHOFF K M,LEATHERS T D,et al.Butyric acid from anaerobic fermentation of lignocellulosic biomass hydrolysates by Clostridium tyrobutyricum strain RPT-4213[J].Bioresource Technology,2013,143:322-329.
- [11]陈玲.饲粮添加丁酸梭菌制剂对断奶仔猪生长性能及免疫的影响[D].雅安:四川农业大学,2018.
- [12]ARSLAN D,STEINBUSCH K J J,DIELS L,et al.Selective short-chain carboxylates production:a review of control mechanisms to direct mixed culture fermentations[J].Critical Reviews in Environmental Science & Technology,2016,46(6):592-634.
- [13]索玉凯.代谢工程改造酪丁酸梭菌(Clostridium tyrobutyricum)强化丁酸生产及木质纤维素利用[D].广州:华南理工大学,2018.
- [14]HUANG J,TANG W,ZHU S Q,et al.Biosynthesis of butyric acid by Clostridium tyrobutyricum[J].Preparative Biochemistry & Biotechnology,2018,48(5):427-434.
- [15]衡文,叶光斌,邹伟.酪丁酸梭菌发酵产丁酸研究进展[J].中国调味品,2021,46(2):186-190.
- [16]马鸿飞,崔宝凯,员瑗,等.响应面法优化褐腐真菌竹生薄孔菌产纤维素酶的液体培养基[J].生物技术通报,2018,34(4):91-101.
- [17]王雪莲,王敏,骆健美,等.枯草芽孢杆菌生防菌B579最佳培养基响应面法优化[J].江苏农业学报,2009,25(1):212-215.
- [18]林华林,秦颖超,周加义,等.丁酸梭菌及其代谢产物对畜禽肠道健康影响的研究进展[J].饲料工业,2019,40(6):11-16.
- [19]赵娜,魏金涛,陈芳,等.丁酸梭菌在动物养殖中的研究与应用[J].湖北畜牧兽医,2018,39(12):13-15.
- [20]戚薇,何玉慧,李安东,等.酪酸梭状芽孢杆菌发酵培养基的优化[J].天津科技大学学报,2010(2):18-21.
- [21]刘磊,田亚男,倪学勤,等.丁酸梭菌 CBM01的碳、氮源优化及其对胃肠道耐受性的研究[J].动物营养学报,2017,29(10):3831-3836.
- [22]亓秀晔,张志焱,谢全喜,等.丁酸梭菌的生物学特性分析及发酵培养基优化[J].中国酿造,2021,40(7):71-76.
- [23]周红,谢全喜.几种丁酸梭菌的性能研究[J].中国酿造,2021,40(3):120-123.
- [24]ABDEL-LATIF M A,EL-HACK M E A.Single and combined effects of Clostridium butyricum and Saccharomyces cerevisiae on growth indices,intestinal health,and immunity of broilers[J].Animals,2018,8(10):184.
- [25]谢丽静,王丽.丁酸梭菌优良菌株LXYB-2抑菌活性及抗逆性研究[J].中国酿造,2018,37(6):91-96.
- [26]杨桂苹.丁酸梭菌的生物学功能研究[J].中国微生态学杂志,1998(5):306-307.