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( Hongyan Yang ),( Hao Wu ),( Lijuan Gao ),( Hongbai Jia ),( Yuan Zhang ),( Zongjun Cui ),( Yuhua Li ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.12
To investigate the effects of Lactobacillus curvatus and Leuconostoc mesenteroides on suan cai (pickled Chinese cabbage) fermentation, L. curvatus and/or Ln. mesenteroides were inoculated into suan cai. Physicochemical indexes were measured, and the microbial dynamics during the fermentation were analyzed by Illumina MiSeq sequencing and quantitative polymerase chain reaction (qPCR). The results showed that inoculation with lactic acid bacteria (LAB) lowered the pH of the fermentation system more rapidly. The decrease in water-soluble carbohydrates in the inoculated treatments occurred more rapidly than in the control. The LAB counts in the control were lower than in other inoculated treatments during the first 12 days of fermentation. According to the Illumina MiSeq sequencing analyses, Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, Cyanobacteria, Fusobacteria, and Verrucomicrobia were present in the fermentations, along with other unclassified bacteria. Generally, Firmicutes was predominant during the fermentation in all treatments. At the genus level, 16 genera were detected. The relative abundance of Lactobacillus in all inoculated treatments was higher than in the control. The relative abundance of Lactobacillus in the treatments containing L. curvatus was higher than in the Ln. mesenteroides-only treatment. The relative abundance of Leuconostoc in the Ln. mesenteroides-containing treatments increased continuously throughout the fermentation. Leuconostoc was highest in the Ln. mesenteroides-only treatment. According to the qPCR results, L. curvatus and/or Ln. mesenteroides inoculations could effectively inhabit the fermentation system. L. curvatus dominated the fermentation in the inoculated treatments.
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Large-scale production of diverse rare ginsenosides using combinatorial biotechnology
Linggai Cao,Hao Wu,He Zhang,Quan Zhao,Xue Yin,Dongran Zheng,Chuanwang Li,Min-Jun Kim,Pyol Kim,Zheyong Xue,Yu Wang,Yuhua Li 강원대학교 산림과학연구소 2019 강원대학교 산림과학연구소 학술대회 Vol.2019 No.09
The rare ginsenosides have superior pharmacological activity than the major ginsenosides. Rare ginsenosides are usually an intermediate product of the ginsenoside biosynthesis pathway and do not accumulate in plants. Therefore, its application in medicine and functional foods is inhibited by the production amount. We developed an effective combinatorial biotechnology approach including large-scale production of ginseng adventitious root in plant bioreactor, hydrolysis of glycosidase combinations and immobilization of glycosidase. Firstly, we used 10 liters to 5 tons of plant bioreactor systems of different sizes to cultivate ginseng adventitious roots, and obtained large amounts of pesticide and heavy metal-free root materials with stable ginsenosides content. The biomass could be increased 30-fold within 60 days and the content of ginsenosides was similar to that of 5-years-old cultivated ginseng. Secondly, we screened out more than ten kinds of glycosidases according to the hydrolysis sites and compared the yield and hydrolysis efficiency of ginsenosides by several combinations of enzymes. Afterwards, a variety of effective combinations were obtained, including Bglsk+Bglmm, BglPm+Bgp1, BglSk+Bgp1, BglSk+BglPm+BgpA. 3g total ginseng saponins extracted from adventitious roots in a 10L bioreactor were converted into more than ten kinds of rare ginsenosides after the enzymes treatment, such as PPD, Rh2, Rg3 and CK. The contents are 336.42 mg, 326.61 mg, 138.54 mg and 279.27 mg, respectively. Thirdly, two kinds of β-glycosidases, BglPm and Bgp1, were immobilized on the hollow fiber membrane. After immobilization, the Km of BglPm and Bgp1 were remain unchanged, while the reaction rates were increased 3-fold and 5-fold, respectively. In the immobilized combination of two glycosidases, the yield of Rh2 was further increased to 511.72 mg. After 2 weeks of immobilization, the β-glycosidases activity remained above 80%. Our combinatorial biotechnology provides an efficient, low-cost, pollution-free and reusable method for large-scale production of diverse rare ginsenosides.
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