食品科学 ›› 2018, Vol. 39 ›› Issue (6): 135-140.doi: 10.7506/spkx1002-6630-201806022

• 生物工程 • 上一篇    下一篇

1 株高产L-乳酸菌株的分离鉴定及其发酵培养基优化

孙丽慧1,王云晓1,吕诗文1,李曼1,贺雷雨2,包永明1   

  1. (1.大连理工大学食品与环境学院,辽宁?盘锦 124221;2.大连理工大学生命与医药学院,辽宁?盘锦 124221)
  • 出版日期:2018-03-25 发布日期:2018-03-14
  • 基金资助:
    中央高校基本科研业务费专项(DUT16QY31)

Isolation and Identification of a Strain for High-Yield Production of L-Lactic Acid and Optimization of Its Culture Medium

SUN Lihui1, WANG Yunxiao1, Lü Shiwen1, LI Man1, HE Leiyu2, BAO Yongming1   

  1. (1. School of Food and Environment, Dalian University of Technology, Panjin 124221, China;2. School of Life Science and Medicine, Dalian University of Technology, Panjin 124221, China)
  • Online:2018-03-25 Published:2018-03-14

摘要: 从辣白菜样品中筛选出1 株高产乳酸的菌株LB-103,经L-/D-乳酸试剂盒检测该菌株发酵产L-乳酸的光学纯度为100%。通过形态学观察、VITEK 2生理生化鉴定和16S rDNA序列分析,确定该菌株为鼠李糖乳酸杆菌(Lactobacillus rhamnosus),将其命名为鼠李糖乳酸杆菌DLF-15038。对其发酵培养基进行初步优化,发现廉价的棉籽饼粉可以部分替代酵母粉,采用15?g/L棉籽饼粉和10?g/L的酵母粉为复合氮源,L-乳酸的产量得以维持且明显降低成本,最适无机盐质量浓度分别为CH3COONa?3?g/L、KH2PO4?2?g/L、MnSO4?0.3?g/L、MgSO4?0.2?g/L。在该优化条件下,进行了5?L发酵罐中的批式流加发酵实验,发酵72?h,L-乳酸产量为165.15?g/L,生产强度为2.29?g/(L·h),糖酸转化率为93.34%。

关键词: L-乳酸, 鼠李糖乳酸杆菌, 分离, 鉴定, 棉籽饼粉

Abstract: A strain, LB-103, which is capable of producing high concentration of L-lactic acid with high optical purity was isolated from kimchi. The optical purity of the produced lactic acid was measured to be 100% by using L-/D-lactic acid enzymatic test kit. The strain was identified as Lactobacillus rhamnosus based on its morphological features, physiological and biochemical properties (VITEK 2 automated microbial identification system), and 16S rDNA sequence analysis. It was named L. rhamnosus DLF-15038. Then the fermentation medium for L-lactic acid production was optimized, and the results showed that yeast extract (10 g/L) and cottonseed meal (15 g/L) were used as nitrogen sources to maintain high L-lactic acid yield and simultaneously reduce the cost. The optimal concentrations of inorganic salts in the medium were as follows: CH3COONa 3 g/L, KH2PO4 2 g/L, MnSO4 0.3 g/L and MgSO4 0.2 g/L. Finally, fed-batch fermentation was performed under the optimized conditions in a 5-L fermentor. We obtained 165.15 g/L L-lactic acid after 72 h fermentation by using the strain DLF-15038, with a yield of 93.34% (g L-lactic acid/g glucose), and a productivity of 2.29 g/(L·h).

Key words: L-lactic acid, Lactobacillus rhamnosus, isolation, identification, cottonseed meal

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