解淀粉芽孢杆菌B17养分需求、发酵条件与绿原酸含量关系的研究

Study on the Relationship between Nutrient Requirements, Fermentation Conditions and Chlorogenic Acid Content of Bacillus amyloliquefaciens B17

绿原酸(chlorogenic acid,CGA)作为一种重要的次生代谢物,在医药、食品、化工等领域广泛应用。本实验室前期研究发现一株产绿原酸的解淀粉芽孢杆菌(Bacillus Amyloliquefaciens)B17,绿原酸产量为1.746 mg/L。为改善该菌株产绿原酸的能力,利用单因素设计优化培养基的培养成分和发酵条件。获得了最佳培养基组分为:蔗糖40 g/L,NH_(4)Cl 5.0 g/L,KH_(2)PO_(4)1.0 g/L,NaCl 3.0 g/L,CuSO_(4)0.025 g/L,FeSO_(4)0.1 g/L,L-苯丙氨酸2 g/L,对羟基苯甲酸10 g/L;并且利用正交设计对发酵条件进行了优化,获得的发酵条件为:温度40℃,pH值7.0,装液量40 mL/250 mL和转速220 r/min。在该条件下,菌株B17产量为1.28 g/L,较优化前产量提高了733.1倍。上述研究结果表明,发酵优化显著提高了菌株B17生产绿原酸的能力,这为后期开发绿原酸在种植、养殖及重金属镉污染等领域的应用提供了原料来源。

Chlorogenic acid(CGA)is widely used as an important secondary metabolite in the fields of medicine,food,and chemical industry.A bacterium strain Bacillus amyloliquefaciens B17 was preserved in our laboratory with a chlorogenic acid production of 1.746 mg/L.A design of single-factor experiment was used to optimize the medium composition and fermentation conditions to improve the production of chlorogenic acid.The optimum medium components were obtained:sucrose 40 g/L,NH_(4)Cl 5.0 g/L,KH_(2)PO_(4)1.0 g/L,NaCl 3.0 g/L,CuSO_(4)0.025 g/L,FeSO_(4)0.1 g/L,L-phenylalanine acid 2 g/L,p-hydroxybenzoic acid 10 g/L.Orthogonal design to optimize the fermentation conditions,the fermentation conditions are:temperature 40℃,pH 7.0,liquid volume 40mL/250 mL and rotational speed 220 r/min.Under this condition,the yield of the strain Bacillus Amyloliquefaciens B17 was 1.28 g/L,which was 733.1 times higher than that before optimization.The above results indicate that the fermentation optimization significantly improved the ability of strain B17 to produce chlorogenic acid,which provides a raw material basis for the later development of CGA application in the fields of planting,breeding and removal of heavy metal cadmium pollution.